Journal Description
DNA
DNA
is an international, peer-reviewed, open access journal on DNA and DNA-related technologies published quarterly online by MDPI.
- Open Access— free for readers, with article processing charges (APC) paid by authors or their institutions.
- Rapid Publication: manuscripts are peer-reviewed and a first decision is provided to authors approximately 23.3 days after submission; acceptance to publication is undertaken in 3.7 days (median values for papers published in this journal in the second half of 2024).
- Recognition of Reviewers: APC discount vouchers, optional signed peer review, and reviewer names published annually in the journal.
Latest Articles
Comparing the Efficiency of Different Methods for Reliable Results in Ancient DNA NGS Workflow
DNA 2025, 5(2), 26; https://doi.org/10.3390/dna5020026 - 19 May 2025
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Background/Objectives: Ancient DNA (aDNA) research workflows heavily depend on efficient aDNA extraction and NGS library preparation. In this study, we compared some of the commonly used laboratory protocols and compared the source of the bone material for sufficient and reliable results. Methods: We
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Background/Objectives: Ancient DNA (aDNA) research workflows heavily depend on efficient aDNA extraction and NGS library preparation. In this study, we compared some of the commonly used laboratory protocols and compared the source of the bone material for sufficient and reliable results. Methods: We executed a three-phase study. First, we analyzed about 2000 previously processed archaic bone samples and conducted a comparative analysis. The second phase involved a controlled experiment of five ancient individuals, with internal control, to further investigate the efficiency of some of the methods. In the third phase, we made a comparison between the efficiency of two enzymes used for library preparation. Results: Samples made from Pars petrosa resulted in the highest yield of endogenous DNA and longer fragment sizes compared to tooth or skeletal samples. DNA extraction made by MinElute columns preserved slightly longer fragments than the handmade silica suspension. NGS libraries indexed using AccuPrime Pfx produced slightly more consistent insert sizes compared to GoTaq G2. Samples prepared with GoTaq G2 contained slightly more unique molecules. The duplication rates showed no significant impact from enzyme choice. Conclusions: Pars petrosa remains the most reliable source of aDNA, with the extraction method using MinElute columns. While AccuPrime Pfx ensures precise NGS library preparation, a more economical choice of the GoTaq G2 enzyme is a viable alternative for degraded archaic samples.
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Open AccessArticle
Synthesis of Sensitive Oligodeoxynucleotides Containing Acylated Cytosine, Adenine, and Guanine Nucleobases
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Komal Chillar, Rohith Awasthy, Marina Tanasova and Shiyue Fang
DNA 2025, 5(2), 25; https://doi.org/10.3390/dna5020025 - 9 May 2025
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Background/Objective: Oligodeoxynucleotides (ODNs) containing base-labile modifications such as N4-acetyldeoxycytidine (4acC), N6-acetyladenosine (6acA), N2-acetylguanosine (2acG), and N4-methyoxycarbonyldeoxycytidine (4mcC) are highly challenging to synthesize because standard ODN synthesis methods require deprotection and cleavage under strongly basic and nucleophilic conditions, and there is a lack of
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Background/Objective: Oligodeoxynucleotides (ODNs) containing base-labile modifications such as N4-acetyldeoxycytidine (4acC), N6-acetyladenosine (6acA), N2-acetylguanosine (2acG), and N4-methyoxycarbonyldeoxycytidine (4mcC) are highly challenging to synthesize because standard ODN synthesis methods require deprotection and cleavage under strongly basic and nucleophilic conditions, and there is a lack of ideal alternative methods to solve the problem. The objective of this work is to explore the capability of the recently developed 1,3-dithian-2-yl-methoxycarbonyl (Dmoc) method for the incorporation of multiple 4acC modifications into a single ODN molecule and the feasibility of using the method for the incorporation of the 6acA, 2acG and 4mcC modifications into ODNs. Methods: The sensitive ODNs were synthesized on an automated solid phase synthesizer using the Dmoc group as the linker and the methyl Dmoc (meDmoc) group for the protection of the exo-amino groups of nucleobases. Deprotection and cleavage were achieved under non-nucleophilic and weakly basic conditions. Results: The 4acC, 6acA, 2acG, and 4mcC were all found to be stable under the mild ODN deprotection and cleavage conditions. Up to four 4acC modifications were able to be incorporated into a single 19-mer ODN molecule. ODNs containing the 6acA, 2acG, and 4mcC modifications were also successfully synthesized. The ODNs were characterized using RP HPLC, capillary electrophoresis, gel electrophoresis and MALDI MS. Conclusions: Among the modified nucleotides, 4acC has been found in nature and proven beneficial to DNA duplex stability. A method for the synthesis of ODNs containing multiple 4acC modifications is expected to find applications in biological studies involving 4acC. Although 6acA, 2acG, and 4mcC have not been found in nature, a synthetic route to ODNs containing them is expected to facilitate projects aimed at studying their biophysical properties as well as their potential for antisense, RNAi, CRISPR, and mRNA therapeutic applications.
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DNA Replication in Time and Space: The Archaeal Dimension
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Anastasia Serdyuk and Thorsten Allers
DNA 2025, 5(2), 24; https://doi.org/10.3390/dna5020024 - 6 May 2025
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The ability of a nucleic acid molecule to self-replicate is the driving force behind the evolution of cellular life and the transition from RNA to DNA as the genetic material. Thus, the physicochemical properties of genome replication, such as the requirement for a
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The ability of a nucleic acid molecule to self-replicate is the driving force behind the evolution of cellular life and the transition from RNA to DNA as the genetic material. Thus, the physicochemical properties of genome replication, such as the requirement for a terminal hydroxyl group for de novo DNA synthesis, are conserved in all three domains of life: eukaryotes, bacteria, and archaea. Canonical DNA replication is initiated from specific chromosomal sequences termed origins. Early bacterial models of DNA replication proposed origins as regulatory points for spatiotemporal control, with replication factors acting on a single origin on the chromosome. In eukaryotes and archaea, however, replication initiation usually involves multiple origins, with complex spatiotemporal regulation in the former. An alternative replication initiation mechanism, recombination-dependent replication, is observed in every cellular domain (and viruses); DNA synthesis is initiated instead from the 3′ end of a recombination intermediate. In the domain archaea, species including Haloferax volcanii are not only capable of initiating DNA replication without origins but grow faster without them. This raises questions about the necessity and nature of origins. Why have archaea retained such an alternative DNA replication initiation mechanism? Might recombination-dependent replication be the ancestral mode of DNA synthesis that was used during evolution from the primordial RNA world? This review provides a historical overview of major advancements in the study of DNA replication, followed by a comparative analysis of replication initiation systems in the three domains of life. Our current knowledge of origin-dependent and recombination-dependent DNA replication in archaea is summarised.
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Unveiling the Genomic Basis of Antagonism and Plant Growth Promotion in the Novel Endophyte Bacillus velezensis Strain B.B.Sf.2
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Dimitra Douka, Tasos-Nektarios Spantidos, Panagiotis Katinakis and Anastasia Venieraki
DNA 2025, 5(2), 23; https://doi.org/10.3390/dna5020023 - 4 May 2025
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Background/Objectives: The agriculture sector faces significant challenges due to global climate change, environmental stressors, and rapid population growth, compounded by unsustainable farming practices. This study investigates the potential of the endophytic bacterial strain B.B.Sf.2, isolated from the bark of Salvia fruticosa and identified
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Background/Objectives: The agriculture sector faces significant challenges due to global climate change, environmental stressors, and rapid population growth, compounded by unsustainable farming practices. This study investigates the potential of the endophytic bacterial strain B.B.Sf.2, isolated from the bark of Salvia fruticosa and identified as Bacillus velezensis through phylogenomic analyses. Methods: To address these issues, eco-friendly techniques, such as the application of plant-associated microbes, are gaining attention. Genome mining revealed numerous secondary metabolite biosynthetic gene clusters associated with plant growth promotion, biocontrol, colonization, and defense elicitation. Results: The strain exhibited strong antagonistic activity against phytopathogens, mediated by diffusible and volatile compound production, along with plant-growth-promoting traits and environmental adaptability. Genome mining revealed numerous secondary metabolite biosynthetic gene clusters associated with plant growth promotion, biocontrol, colonization, and defense elicitation. B.B.Sf.2 effectively inhibited Colletotrichum species causing olive anthracnose and suppressed Botrytis cinerea, the gray mold pathogen, in post-harvest studies on infected fruits. Bioautography of ethyl acetate extracts demonstrated bioactivity against B. cinerea, attributed to iturin-like metabolites. The extracts maintained bioactive properties regardless of fungal interaction. Furthermore, the strain significantly promoted the growth of Arabidopsis thaliana via diffusible and volatile compounds. Conclusions: Our results highlight the multifunctional potential of B.B.Sf.2 as a biocontrol and growth-promoting agent, warranting further evaluation in field applications to enhance sustainable agriculture.
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Open AccessReview
Genomic and Epigenomic Plasticity in the Hypoxic Environment
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Emanuele Cricchi, Alessio Bertozzo, Martina Minisini and Claudio Brancolini
DNA 2025, 5(2), 22; https://doi.org/10.3390/dna5020022 - 4 May 2025
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Our genome has evolved a complex network of information designed to precisely regulate gene transcription. Commonly known as cis-regulatory elements, they represent those parts of DNA that are highly sensitive to environmental changes in the form of associated multi-protein complexes. Oxygen levels are
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Our genome has evolved a complex network of information designed to precisely regulate gene transcription. Commonly known as cis-regulatory elements, they represent those parts of DNA that are highly sensitive to environmental changes in the form of associated multi-protein complexes. Oxygen levels are an important environmental factor influencing a range of cellular activities, including cell survival. To respond to changes in oxygen levels, cells have developed an efficient and precise system for regulating gene expression. Cis-regulatory elements are the key hubs of this response and control the activation of the transcriptional response to hypoxia. In this review, we will discuss the complex genomic and epigenomic structures that are modulated by oxygen and control the activity of cis-regulatory elements and the adaptations to variations in O2 availability.
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(This article belongs to the Special Issue Epigenetics and Environmental Exposures)
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Research Themes in KAT6A Syndrome: A Scoping Review
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Tanya Tripathi, Miya St John, Jordan Wright, Natacha Esber and David J. Amor
DNA 2025, 5(2), 21; https://doi.org/10.3390/dna5020021 - 27 Apr 2025
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Pathogenic variants in the KAT6A gene cause KAT6A syndrome, a neurodevelopmental disorder characterised by intellectual disability (ID), developmental delay, speech and language challenges, feeding difficulties, and skeletal abnormalities. This scoping review synthesises current knowledge on KAT6A syndrome, identifies key research themes, and supports
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Pathogenic variants in the KAT6A gene cause KAT6A syndrome, a neurodevelopmental disorder characterised by intellectual disability (ID), developmental delay, speech and language challenges, feeding difficulties, and skeletal abnormalities. This scoping review synthesises current knowledge on KAT6A syndrome, identifies key research themes, and supports the mission of advocacy groups like the KAT6 Foundation. A systematic search of five databases (Ovid MEDLINE, Ovid EMBASE, PubMed, Web of Science, and Scopus) was conducted from 1990 to 2024, including peer-reviewed articles, preprints, and conference abstracts published from 2022 onward. Of 771 citations retrieved, 111 full-text articles were reviewed, with 62 meeting the inclusion criteria. Data were synthesised into six themes: (1) the genotype and phenotype map, revealing a broad phenotypic spectrum with common features like ID, absent speech, and craniofacial dysmorphism, as well as rare features such as severe aplastic anaemia and pancraniosynostosis; (2) the neurodevelopmental profile, detailing communication deficits, sleep disturbances, and impaired adaptive functioning; (3) the epigenetic and developmental roles of KAT6A, highlighting its critical function in histone acetylation, chromatin remodelling, and gene regulation; (4) molecular biomarkers, identifying distinct DNA methylation episignatures and dysregulated cellular pathways; (5) drug discovery, with preliminary studies suggesting that pantothenate and L-carnitine may mitigate mitochondrial dysfunction and histone acetylation deficits, while RSPO2 overexpression reverses cognitive impairment in animal models; (6) phenotypic overlap with Rett syndrome and KAT6B-related disorders. This review underscores the complexity and variability of KAT6A syndrome, highlighting the need for multidisciplinary approaches to improving diagnosis, management, and development of therapies. Future research should focus on longitudinal studies, underrepresented phenotypes, biomarker identification, and robust therapeutic trials to enhance outcomes for affected individuals and their families.
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Open AccessArticle
DNA Barcoding as a Tool for Surveying Cytospora Species Associated with Branch Dieback and Canker Diseases of Woody Plants in Canada
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Evgeny Ilyukhin and Svetlana Markovskaja
DNA 2025, 5(2), 20; https://doi.org/10.3390/dna5020020 - 21 Apr 2025
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Background/Objectives: Branch dieback and canker diseases caused by Cytospora species adversely impact the health of woody plants worldwide. Results: During this survey, 59 Cytospora isolates were obtained from symptomatic trees and shrubs growing in southwest Ontario and Saskatchewan, Canada. A DNA barcoding approach
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Background/Objectives: Branch dieback and canker diseases caused by Cytospora species adversely impact the health of woody plants worldwide. Results: During this survey, 59 Cytospora isolates were obtained from symptomatic trees and shrubs growing in southwest Ontario and Saskatchewan, Canada. A DNA barcoding approach combined with morphological characterization identified 15 known species of Cytospora associated with these diseases: C. chrysosperma, C. curvata, C. euonymina, C. hoffmannii, C. kantschavelii, C. leucosperma, C. leucostoma, C. nitschkeana, C. piceae, C. populina, C. pruinopsis, C. pruinosa, C. ribis, C. schulzeri, and C. sorbina. The most common species isolated from multiple hosts were C. sorbina (10), C. chrysosperma (8), C. nitschkeana (6), and C. pruinosa (6). A wide range of host associations, including non-conifer species, was observed for C. piceae. Conclusions: The obtained results contribute to the study of diversity, host affiliation, geographical distribution, and pathogenicity of Cytospora species occurring on woody plants in both natural habitats and agricultural systems. The findings support the effectiveness of using DNA barcodes in fungal taxonomy and plant pathology studies.
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Open AccessReview
Chemical Versus Enzymatic Nucleic Acid Modifications and Genomic Stability
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Jonathan R. Cortez and Marie E. Migaud
DNA 2025, 5(2), 19; https://doi.org/10.3390/dna5020019 - 9 Apr 2025
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DNA damage and repair have been central themes in cellular biology research. Broadly, DNA damage is understood as modifications to canonical nucleotides that disrupt their function during transcription and replication. A deeper biochemical understanding of DNA damage is essential, as the genome governs
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DNA damage and repair have been central themes in cellular biology research. Broadly, DNA damage is understood as modifications to canonical nucleotides that disrupt their function during transcription and replication. A deeper biochemical understanding of DNA damage is essential, as the genome governs all cellular processes. We can classify DNA damage according to whether the modifications to the nucleic acid scaffold are chemically or enzymatically initiated. This distinction is important because chemical modifications are often irreversible, sometimes sparse, and difficult to detect or control spatially and replicate systematically. This can result in genomic damage or modifications to nucleotides in the nucleotide pool, which is less commonly studied. In contrast, enzymatic modifications are typically induced by the cell for specific purposes and are under strong regulatory control. Enzymatic DNA modifications also present a degree of sequence specificity and are often reversible. However, both types of DNA modifications contribute to cellular aging when poorly repaired and, as a result, remain incompletely understood. This review hopes to gather less studied mechanisms in nucleotide modifications and show research gaps in our current understanding of nucleotide biology. By examining the implications of these mechanisms on DNA modifications, in the nucleotide pool and genome, we may gain insights into innovative strategies for mitigating the effects of cellular aging.
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(This article belongs to the Special Issue Epigenetics and Environmental Exposures)
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Thyroid Hormone-Responsive Genes in Primary Cultures of Rat Hepatic Cells
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Nariaki Fujimoto and Shigeyuki Kitamura
DNA 2025, 5(2), 18; https://doi.org/10.3390/dna5020018 - 1 Apr 2025
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Background/Objectives: Thyroid hormones are key regulators in hepatic metabolic pathways. Although they regulate various hepatic genes, only a few are known to be under direct transcriptional regulation through thyroid hormone receptors. To better understand the roles of thyroid hormones in the liver, it
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Background/Objectives: Thyroid hormones are key regulators in hepatic metabolic pathways. Although they regulate various hepatic genes, only a few are known to be under direct transcriptional regulation through thyroid hormone receptors. To better understand the roles of thyroid hormones in the liver, it is critical to identify thyroid hormone-responsive genes at the cellular level. Methods: A cDNA microarray analysis was applied to primary cultures of rat hepatic cells treated with triiodothyronine (T3) at 10−9 M for 24 h to identify the differentially expressed genes. The identified gene expressions were further examined in vivo using F344 rats. The reporter gene assay was performed to investigate the transcriptional activity of the upstream region of the gene. Results: A limited number of genes were listed, and only three of them, pyridoxal kinase (Pdxk), phosphoenolpyruvate carboxykinase 1 (Pck1), and solute carrier family 17 member 2 (Slc17a2), were confirmed to be upregulated by quantitative RT-PCR. The mRNA expression of these genes increased in the livers of F344 rats after T3 injection, suggesting the physiological relevance in vivo. There are two partially conserved thyroid hormone-responsive elements (TREs) in the upstream region of the rat Pdxk gene. The reporter gene assay indicated that an imperfect TRE (5′-gGGTCAxxxxAGGaCt-3′) located at −2146 was sufficient for the thyroid hormone-induced transcription of the gene. Conclusions: The present study identified novel T3-responsive genes, pdxk and Slc17a2. Promoter analyses showed that a single TRE in the pdxk gene accounts for the transcriptional regulation by T3.
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Overview of Roles of Novel Components in the Regulation of DNA Damage Repair in BRCA1-Deficient Cancers: An Update
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Nhat Nguyen, Dominic Arris and Manh Tien Tran
DNA 2025, 5(2), 17; https://doi.org/10.3390/dna5020017 - 1 Apr 2025
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Cancers that arise from germline mutations of breast cancer associated gene 1 (BRCA1), which is a crucial player in homologous recombination (HR) DNA repair, are vulnerable to DNA-damaging agents such as platinum and PARP inhibitors (PARPis). Increasing evidence suggests that BRCA1
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Cancers that arise from germline mutations of breast cancer associated gene 1 (BRCA1), which is a crucial player in homologous recombination (HR) DNA repair, are vulnerable to DNA-damaging agents such as platinum and PARP inhibitors (PARPis). Increasing evidence suggests that BRCA1 is an essential driver of all phases of the cell cycle, thereby maintaining orderly steps during cell cycle progression. Specifically, loss of BRCA1 activity causes the S-phase, G2/M, spindle checkpoints, and centrosome duplication to be dysregulated, thereby blocking cell proliferation and inducing apoptosis. In vertebrates, loss of HR genes such as BRCA1 and/or BRCA2 is lethal, since HR is a prerequisite for genome integrity. Thus, cancer cells utilize alternative DNA repair pathways such as non-homologous end joining (NHEJ) to cope with the loss of BRCA1 function. In this review, we attempt to update and discuss how these novel components are crucial for regulating DNA damage repair (DDR) in BRCA1-deficient cancers.
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BUB1 Inhibition Induces Ferroptosis in Triple-Negative Breast Cancer Cell Lines
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Sushmitha Sriramulu, Shivani Thoidingjam, Stephen L. Brown, Farzan Siddiqui, Benjamin Movsas and Shyam Nyati
DNA 2025, 5(1), 16; https://doi.org/10.3390/dna5010016 - 12 Mar 2025
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Background: Triple-negative breast cancer (TNBC) is a highly aggressive subtype with limited effective treatments available, including targeted therapies, often leading to poor prognosis. Mitotic checkpoint kinase BUB1 is frequently overexpressed in TNBC and correlates with poor survival outcomes suggesting its potential as
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Background: Triple-negative breast cancer (TNBC) is a highly aggressive subtype with limited effective treatments available, including targeted therapies, often leading to poor prognosis. Mitotic checkpoint kinase BUB1 is frequently overexpressed in TNBC and correlates with poor survival outcomes suggesting its potential as a therapeutic target. This study explores the cytotoxicity of TNBC cells to BUB1 inhibition, alone or in combination with radiation and demonstrates that ferroptosis, an iron-dependent form of programmed cell death, has a role. Methods: TNBC cell lines (SUM159, MDA-MB-231, and BT-549) were treated with a BUB1 inhibitor BAY1816032 (BUB1i) alone or in combination with the ferroptosis activator RSL3 with or without 4 Gy irradiation. Cell viability assays were conducted to assess treatment effects, qPCR analyses measured expression of key ferroptosis markers including ACSL4, GPX4, PTGS2, SLC7A11, NCOA4, IREB2, NFS1, and TFRC expression, and TBARS assay measured the lipid peroxidation levels. Ferroptosis specificity was confirmed through co-treatment with the ferroptosis inhibitor Ferrostatin-1 (F-1). Results: In all TNBC cell lines studied, BUB1 inhibition significantly induced ferroptosis, marked by increased expression of ACSL4 and PTGS2, decreased expression of GPX4 and SLC7A11, and increased lipid peroxidation levels. The combination of BUB1i with RSL3 further amplified these ferroptotic markers, suggesting at least an additive effect, which was not present with the combination of BUB1i and radiation. Co-treatment with Ferrostatin-1 reversed the expression of ferroptosis markers, suggesting that BUB1i-mediated cell death may involve ferroptotic signaling in TNBC cell lines. Conclusions: This study demonstrates that BUB1 inhibition may independently induce ferroptosis in TNBC cell lines, which is enhanced when combined with a ferroptosis activator. Further research is warranted to delineate the molecular mechanism of BUB1-mediated ferroptosis in TNBC.
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Open AccessArticle
Carbon Dioxide Fluxes Associated with Prokaryotic and Eukaryotic Communities in Ice-Free Areas on King George Island, Maritime Antarctica
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Luiz H. Rosa, Vívian N. Gonçalves, Débora Luiza Costa Barreto, Marcio Rocha Francelino, Clara Glória Oliveira Baldi, Danilo Cesar Mello, Kárita C. R. Santos, Fabyano A. C. Lopes, Micheline Carvalho-Silva, Peter Convey and Paulo E. A. S. Câmara
DNA 2025, 5(1), 15; https://doi.org/10.3390/dna5010015 - 10 Mar 2025
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Background and Methods: We assessed the prokaryotic and eukaryotic diversity present in non-vegetated and vegetated soils on King George Island, Maritime Antarctic, in combination with measurements of carbon dioxide fluxes. Results: For prokaryotes, 381 amplicon sequence variants (ASVs) were assigned, dominated by the
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Background and Methods: We assessed the prokaryotic and eukaryotic diversity present in non-vegetated and vegetated soils on King George Island, Maritime Antarctic, in combination with measurements of carbon dioxide fluxes. Results: For prokaryotes, 381 amplicon sequence variants (ASVs) were assigned, dominated by the phyla Actinobacteriota, Acidobacteriota, Pseudomonadota, Chloroflexota, and Verrucomicrobiota. A total of 432 eukaryotic ASVs were assigned, including representatives from seven kingdoms and 21 phyla. Fungi dominated the eukaryotic communities, followed by Viridiplantae. Non-vegetated soils had higher diversity indices compared with vegetated soils. The dominant prokaryotic ASV in non-vegetated soils was Pyrinomonadaceae sp., while Pseudarthrobacter sp. dominated vegetated soils. Mortierella antarctica (Fungi) and Meyerella sp. (Viridiplantae) were dominant eukaryotic taxa in the non-vegetated soils, while Lachnum sp. (Fungi) and Polytrichaceae sp. (Viridiplantae) were dominant in the vegetated soils. Measured CO2 fluxes indicated that the net ecosystem exchange values measured in vegetated soils were lower than ecosystem respiration in non-vegetated soils. However, the total flux values indicated that the region displayed positive ecosystem respiration values, suggesting that the soils may represent a source of CO2 in the atmosphere. Conclusions: Our study revealed the presence of rich and complex communities of prokaryotic and eukaryotic organisms in both soil types. Although non-vegetated soils demonstrated the highest levels of diversity, they had lower CO2 fluxes than vegetated soils, likely reflecting the significant biomass of photosynthetically active plants (mainly dense moss carpets) and their resident organisms. The greater diversity detected in exposed soils may influence future changes in CO2 flux in the studied region, for which comparisons of non-vegetated and vegetated soils with different microbial diversities are needed. This reinforces the necessity for studies to monitor the impact of resident biota on CO2 flux in different areas of Maritime Antarctica, a region strongly impacted by climatic changes.
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Open AccessEditor’s ChoiceArticle
Conservation Genomics of West Virginia Walleye (Sander vitreus): Impact of Minor Allele Frequency Thresholds on Population Structure and Potential Adaptive Divergence Inferences
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Andrew Johnson, Katherine Zipfel, Dustin Smith and Amy Welsh
DNA 2025, 5(1), 14; https://doi.org/10.3390/dna5010014 - 3 Mar 2025
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Background: Walleye (Sander vitreus), a valuable sportfish and an important ecological apex predator, exhibits genetic structuring across their range and localized structuring as a result of stocking. Methods: Walleye from 17 sampling locations across West Virginia were sequenced using a ddRAD
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Background: Walleye (Sander vitreus), a valuable sportfish and an important ecological apex predator, exhibits genetic structuring across their range and localized structuring as a result of stocking. Methods: Walleye from 17 sampling locations across West Virginia were sequenced using a ddRAD protocol, generating various SNP datasets to assess population structuring and genomic diversity, with specific emphasis on the native Eastern Highlands strain. Different minor allele frequency filter thresholds were tested to assess impacts on genetic diversity and differentiation metrics. Results: High genetic differentiation was observed between the Eastern Highlands and Great Lakes strains, with further sub-structuring within the Eastern Highlands strain between the Ohio River populations and the other populations. Increasing MAF thresholds generally reduced the distinctiveness of clusters, but the overall inference of the number of clusters was minimally impacted. Genetic diversity metrics indicated some variability among Eastern Highlands walleye populations, with isolated populations, including the New River and Summersville Lake, showing higher inbreeding coefficients. MAF filters generally increased diversity metrics, but the trend of diversity metrics among populations remained relatively consistent. Several SNPs were found to be potentially undergoing selection, with the minor allele frequencies of these SNPs being found to be highest in Summersville Lake, highlighting potential adaptive divergence between the riverine populations and a large lentic system. Conclusions: The use of any MAF filter generated the same trends of population structuring and genomic diversity inferences regardless of the MAF threshold used. Further management of Eastern Highlands walleye in West Virginia needs to emphasize protecting the genetic integrity of the Kanawha River population and ongoing genomic screening of broodstock to conserve native genetic diversity.
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Open AccessEditor’s ChoiceArticle
Estimating Carbon Biomass Using DNA: Phytoplankton as a Case Study
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Lingjie Zhou, Nanjing Ji, Brittany N. Sprecher and Senjie Lin
DNA 2025, 5(1), 13; https://doi.org/10.3390/dna5010013 - 3 Mar 2025
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Background/Objectives: Estimating carbon content for cells is often necessary but difficult. In many biological, oceanographic, and marine biogeochemical studies, information on phytoplankton species composition and their biomass contribution to the community is essential. However, it is technically challenging to estimate the biomass of
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Background/Objectives: Estimating carbon content for cells is often necessary but difficult. In many biological, oceanographic, and marine biogeochemical studies, information on phytoplankton species composition and their biomass contribution to the community is essential. However, it is technically challenging to estimate the biomass of individual species in a natural assemblage. DNA analysis has the potential to profile species composition and estimate species-specific carbon biomass simultaneously. However, this requires an established relationship between carbon biomass and DNA content with species resolution using a measurable DNA index such as rDNA. Methods: In this study, DNA, rDNA, and carbon contents were measured for species from major phytoplankton phyla grown in different growth stages and under different nutrient and temperature conditions. Correlations between these parameters were examined. Results: Our data resulted in significant log-log regression equations: Log C = 0.8165 × Log DNA + 2.407 (R2 = 0.9577, p < 0.0001), Log rDNA = 0.7472 × Log DNA − 0.0289 (R2 = 0.9456, p < 0.0001), and Log C = 1.09 × Log rDNA + 2.41 (R2 = 0.9199, p < 0.0001). Furthermore, similar strong regression functions were found when incorporating previously published data on a wide range of organisms including bacteria, plants, and animals. Conclusions: Carbon biomass is significantly correlated with DNA and rDNA abundances in phytoplankton and other organisms. The regression equations we developed offer a tool for estimating phytoplankton carbon biomass using DNA or rDNA and serve as a foundation for establishing similar models for other organisms.
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Open AccessEditor’s ChoiceArticle
Peroxidase-like Activity of G-Quadruplex/Hemin Complexes for Colorimetric Nucleic Acid Analysis: Loop and Flanking Sequences Affect Signal Intensity
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Ryan P. Connelly, Valentina Fonseca and Yulia V. Gerasimova
DNA 2025, 5(1), 12; https://doi.org/10.3390/dna5010012 - 3 Mar 2025
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Background/Objectives: Some G-quadruplex (G4)-forming nucleic acid sequences bind a hemin cofactor to enhance its peroxidase-like activity. This has been implemented in a variety of bioanalytical assays benefiting from analyte-dependent peroxidation of a chromogenic organic substrate (e.g., ABTS) to produce a color change.
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Background/Objectives: Some G-quadruplex (G4)-forming nucleic acid sequences bind a hemin cofactor to enhance its peroxidase-like activity. This has been implemented in a variety of bioanalytical assays benefiting from analyte-dependent peroxidation of a chromogenic organic substrate (e.g., ABTS) to produce a color change. Adenine and cytosine nucleotides in the vicinity of the G4 hemin-binding site promote the peroxidation reaction. In this work, the effect of G4 loop and flanking nucleotides on the colorimetric signal of split hybridization probes utilizing hemin-G4 signal reporters was tested. Methods: G4s varying by loop sequences and flanking nucleotides were tested with hemin for ABTS peroxidation (A420), and the signal was compared with that produced by the most catalytically efficient complexes reported in the literature using one-way ANOVA and post hoc pairwise comparison with Tukey’s HSD test. The best G4s were used as signal transducers in the split peroxidase deoxyribozyme (sPDz) probes for sensing two model nucleic acid analytes, as well as in a cascade system, where the analyte-dependent assembly of an RNA-cleaving deoxyribozyme 10–23 results in G4 release. Results: Intramolecular G4s (G3T)3G3TC or G3T3G3ATTG3T3G3 were found to be the most efficient hemin PDzs. When splitting intramolecular G4 for the purpose of sPDz probe design, the addition of a flanking d(TC) sequence at one of the G4 halves or d(ATT) in a loop connecting the second and third G-tracts helps boost analyte-dependent signal intensity. However, for the cascade system, the effect of d(TC) or d(ATT) in the released G4 was not fully consistent with the data reported for intramolecular G4-hemin complexes. Conclusions: Our findings offer guidance on the design of split hybridization probes utilizing the peroxidase-like activity of G4-hemin complexes as a signal transducer.
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Open AccessEditor’s ChoiceArticle
Similar Ehlers–Danlos Syndrome Profiles Produced by Variants in Multiple Collagen Genes
by
Sahil S. Tonk and Golder N. Wilson
DNA 2025, 5(1), 11; https://doi.org/10.3390/dna5010011 - 25 Feb 2025
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Background: Despite increased attention to double-jointedness or joint hypermobility as seen in connective tissue dysplasias like Ehlers–Danlos syndrome, improved clinical DNA correlations are needed to reduce decadal delays in diagnosis. Methods: To this end, patterns of history (among 80) and physical (among 40)
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Background: Despite increased attention to double-jointedness or joint hypermobility as seen in connective tissue dysplasias like Ehlers–Danlos syndrome, improved clinical DNA correlations are needed to reduce decadal delays in diagnosis. Methods: To this end, patterns of history (among 80) and physical (among 40) findings are compared for 121 Ehlers–Danlos syndrome patients with recurring variants in collagen type I, II, III, V, VI, VII, IX, XI, and XII genes and novel ones in type XV, XVII, XVIII, and XXVII. Results: A recognizable tissue laxity–dysautonomia profile that transcended collagen biochemical class, triple helix component, mutation type, or presence of accessory DNA variants was defined with a few exceptions. Patients with novel variations experienced severe symptoms at younger ages (6–10 versus 14–18 years) and those with collagen type III variations had more than one significant difference in finding frequencies (spinal disk issues 75% versus 49%; bloating-reflux 100% versus 69%; migraines or menorrhagia 92% versus 53%). Conclusions: These results suggest that collagen DNA variants of diverse gene and molecular type can demonstrate EDS disposition and hasten its diagnosis when distress and disease become manifest.
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Open AccessEditor’s ChoiceReview
Essays on the Binary Representations of the DNA Data
by
Evgeny V. Mavrodiev and Nicholas E. Mavrodiev
DNA 2025, 5(1), 10; https://doi.org/10.3390/dna5010010 - 14 Feb 2025
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The advancement of modern genomics has led to the large-scale industrial production of molecular data and scientific outcomes. Simultaneously, conventional DNA character alignments (sequence alignments) are utilized for DNA-based phylogenetic analyses without further recoding procedures or any a priori determination of character polarity,
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The advancement of modern genomics has led to the large-scale industrial production of molecular data and scientific outcomes. Simultaneously, conventional DNA character alignments (sequence alignments) are utilized for DNA-based phylogenetic analyses without further recoding procedures or any a priori determination of character polarity, contrary to the requirements of foundations of phylogenetic systematics. These factors are the primary reasons why the binary perspective has not been implemented in modern molecular phylogenetics. In this study, we demonstrate how to recode conventional DNA data into various types of binary matrices, either unpolarized or with established polarity. Despite its historical foundation, our analytical approach to DNA sequence data has not been adequately explored since the inception of the molecular age. Binary representations of conventional DNA alignments allow for the analysis of molecular data from a purely comparative or static perspective. Furthermore, we show that the binarization of DNA data possesses broad mathematical and cultural connotations, making them intriguing regardless of their applications to different phylogenetic procedures.
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Open AccessFeature PaperEditor’s ChoiceReview
Contribution of Androgen Receptor CAG Repeat Polymorphism to Human Reproduction
by
Alessandro Ciarloni, Nicola delli Muti, Nicola Ambo, Michele Perrone, Silvia Rossi, Sara Sacco, Gianmaria Salvio and Giancarlo Balercia
DNA 2025, 5(1), 9; https://doi.org/10.3390/dna5010009 - 8 Feb 2025
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Background: Exon 1 of the gene encoding for the androgen receptor (AR) contains a polymorphic sequence of variably repeated CAG triplets ranging from 11 to 36. The number of triplets appears to inversely correlate with receptor transcriptional activity, conditioning the peripheral effects
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Background: Exon 1 of the gene encoding for the androgen receptor (AR) contains a polymorphic sequence of variably repeated CAG triplets ranging from 11 to 36. The number of triplets appears to inversely correlate with receptor transcriptional activity, conditioning the peripheral effects of testosterone. Methods: We conducted a narrative review to explore the current evidence regarding the relationship between the number of CAG repeats and the human reproductive system. Results: We found several articles that investigate the relationship between CAG polymorphism and the male reproductive system, suggesting a possible modulatory effect on spermatogenesis, sexual function, prostate cancer, and testicular cancer. Similarly, in women, evidence has emerged to support a possible relationship between CAG repeat number and breast cancer, polycystic ovary syndrome (PCOS), and recurrent spontaneous abortions (RSAs). Unfortunately, the data in the current literature are largely discordant, largely due to an important influence of ethnicity on the variability of the CAG polymorphism, and partly due to the quality of the available studies. Conclusions: In the current state of the art, the study of CAG polymorphism does not have a sufficient literature base to allow its use in common clinical practice. However, it represents an interesting research target and, in the future, as new evidence emerges, it could help to elucidate some pathogenetic aspects of human reproductive disorders.
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Open AccessEditor’s ChoiceArticle
Transcription Factor Inhibition as a Potential Additional Mechanism of Action of Pyrrolobenzodiazepine (PBD) Dimers
by
Julia Mantaj, Paul J. M. Jackson, Richard B. Parsons, Tam T. T. Bui, David E. Thurston and Khondaker Miraz Rahman
DNA 2025, 5(1), 8; https://doi.org/10.3390/dna5010008 - 5 Feb 2025
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Background: The pyrrolobenzodiazepine (PBD) dimer SJG-136 reached Phase II clinical trials in ovarian cancer and leukaemia in the UK and USA in the 2000s. Several structural analogues of SJG-136 are currently in clinical development as payloads for Antibody-Drug Conjugates (ADCs). There is growing
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Background: The pyrrolobenzodiazepine (PBD) dimer SJG-136 reached Phase II clinical trials in ovarian cancer and leukaemia in the UK and USA in the 2000s. Several structural analogues of SJG-136 are currently in clinical development as payloads for Antibody-Drug Conjugates (ADCs). There is growing evidence that PBDs exert their pharmacological effects through inhibition of transcription factors (TFs) in addition to arrest at the replication fork, DNA strand breakage, and inhibition of enzymes including endonucleases and RNA polymerases. Hence, PBDs can be used to target specific DNA sequences to inhibit TFs as a novel anticancer therapy. Objective: To explore the ability of SJG-136 to bind to the cognate sequences of transcription factors using a previously described HPLC/MS method, to obtain preliminary mechanistic evidence of its ability to inhibit transcription factors (TF), and to determine its effect on TF-dependent gene expression. Methods: An HPLC/MS method was used to assess the kinetics and thermodynamics of adduct formation between the PBD dimer SJG-136 and the cognate recognition sequence of the TFs NF-κB, EGR-1, AP-1, and STAT3. CD spectroscopy, molecular dynamics simulations, and gene expression analyses were used to rationalize the findings of the HPLC/MS study. Results: Notable differences in the rate and extent of adduct formation were observed with different DNA sequences, which might explain the variations in cytotoxicity of SJG-136 observed across different tumour cell lines. The differences in adduct formation result in variable downregulation of several STAT3-dependent genes in the human colon carcinoma cell line HT-29 and the human breast cancer cell line MDA-MB-231. Conclusions: SJG-136 can disrupt transcription factor-mediated gene expression, which contributes to its exceptional cytotoxicity in addition to the DNA-strand cleavage initiated by its ability to crosslink DNA.
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Open AccessEditor’s ChoiceArticle
Partial Proliferating Cell Nuclear Antigen Functional Knockout Impairs Cisplatin Resistance and Clonogenic Potential in Lung Adenocarcinoma Cells
by
Ana Paula Morelli, Nathalia Quintero-Ruiz, Mariana Camargo Silva Mancini, Isadora Carolina Betim Pavan, Isabelle Lima Flores, Luiz Guilherme Salvino Silva, Matheus Brandemarte Severino, Rosangela Maria Neves Bezerra and Fernando Moreira Simabuco
DNA 2025, 5(1), 7; https://doi.org/10.3390/dna5010007 - 2 Feb 2025
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Background/Objectives: Lung cancer ranks as the leading cause of cancer-related deaths globally and is highly associated with cisplatin resistance due to both intrinsic and extrinsic mechanisms. Proliferating Cell Nuclear Antigen (PCNA) plays a critical role in molecular processes, such as DNA replication and
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Background/Objectives: Lung cancer ranks as the leading cause of cancer-related deaths globally and is highly associated with cisplatin resistance due to both intrinsic and extrinsic mechanisms. Proliferating Cell Nuclear Antigen (PCNA) plays a critical role in molecular processes, such as DNA replication and repair, chromatin structure maintenance, and cell cycle progression. PCNA is known as a molecular marker for proliferation and an excellent inhibition target to shut down highly proliferative cells. One of the mechanisms of cisplatin resistance is the increase in DNA repair, and studies have reported an association between PCNA, lung cancer, and cisplatin treatment. The present study aimed to characterize the absence of PCNA in A549 lung adenocarcinoma cells. Methods: Employing a CRISPR/Cas9 gene-editing approach, we generated a monoclonal cell culture, termed PKO (PCNA knockout). Results: PKO cells exhibited a residual PCNA expression, significantly decreased clonogenic potential and ubiquitylation at K164 residue. IC50 assay suggested that PKO cells could not acquire cisplatin resistance when compared to PX. After cisplatin treatment, PKO cells presented impaired ubiquitylation and did not have increased STAT3 phosphorylation (Tyr705), a previously characterized mechanism of cisplatin resistance. Conclusions: We suggest that PCNA participates in cisplatin resistance in A549, partially by DNA damage tolerance through failure on PCNA monoubiquitylation, and its inhibition may be an approach to circumvent cisplatin resistance.
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