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Special Issue "Detection and Safety Assessment of Genetically Modified Organisms"

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A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Molecular Toxicology".

Deadline for manuscript submissions: closed (15 August 2014)

Special Issue Editor

Guest Editor
Dr. Virginia Garcia-Cañas

Laboratory of Foodomics (CIAL), National Research Council of Spain (CSIC), Nicolás Cabrera 9, 28049, Madrid, Spain
Website | E-Mail
Interests: food analysis; bioactivity of dietary ingredients; foodomics; high-throughput analysis; transcriptomics; genomics; proteomics; in vitro biological models

Special Issue Information

Dear Colleagues,

Since its early applications more than three decades ago, genetic engineering has become one of the principal technological advances in modern biotechnology. The rapid progress of this technology has opened new opportunities to create genetically modified organisms (GMOs), which are used to grow GM crops for the production of food, feed, etc. However, public attention to the use of genetic engineering in the production of GMOs has been constantly growing since its beginning. The main controversial issues focus on four areas, namely, potential harm to human health, environmental concerns, and concerns related to ethical and patent issues.

After several years of intense debate, regulations on different aspects regarding GMOs, including risk assessment, marketing, labeling, and traceability have been established in many countries. In order to verify compliance with the requirements imposed by the legislation, appropriate analytical tools have been developed and validated for the rapid detection, identification and accurate quantification of approved and unapproved GMOs in food and feed. However, the variety of authorized and unauthorized GMOs and the extensive areas where they are cultivated are steadily increasing around the globe, making their detection more challenging. In consequence, there remains a demand for innovative analytical procedures able to cope with the increasingly growing number of novel GMOs in a cost-effective and rapid way.

Safety evaluation of GMO-derived foods is based on the comparative analysis between the GMO and its conventional counterpart in terms of allergenicity, toxicity, nutrients, etc. Also, the study of potential unintended alterations, defined as those effects that fall beyond the primary expected effects of the genetic modification, has attracted much attention in recent years. As they are unpredictable, unintended effects are considered as a source of uncertainty that might pose health risks. Although their detection is challenging, different analytical approaches, including targeted analysis and the more recent profiling strategies based on omics techniques, have been developed in recent years. In spite of these developments, more robust, reliable and integrative approaches that facilitate comprehensive or global compositional studies to effectively investigate safety and quality aspects of GMOs are lacking. In this context, appropriate holistic strategies such as foodomics are promising approaches to tackle the high level of complexity in the study of GMOs.

A special issue devoted to Detection and Safety Assessment of Genetically Modified Organisms in the International Journal of Molecular Sciences is thus timely and well-grounded. All manuscripts furthering advanced methodologies for the analysis of GMOs in food and feed samples are welcome. Of particular interest would be those works that focus on the development and application of high-throughput technologies that allow cost-effective, rapid and/or multi-targeted GMO analysis. Studies based on omics profiling and foodomics, aimed at investigating safety and quality issues are also called for.

Dr. Virginia Garcia-Cañas
Guest Editor

Submission

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. Papers will be published continuously (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as communications are invited. For planned papers, a title and short abstract (about 100 words) can be sent to the Editorial Office for announcement on this website.

Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are refereed through a peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. International Journal of Molecular Sciences is an international peer-reviewed Open Access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 1600 CHF (Swiss Francs).


Keywords

  • genetically modified organisms
  • transgenic food
  • molecular methods
  • unintended effects
  • foodomics
  • DNA analysis
  • safety assessment
  • omics technologies

Published Papers (7 papers)

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Research

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Open AccessArticle Mutation Scanning in a Single and a Stacked Genetically Modified (GM) Event by Real-Time PCR and High Resolution Melting (HRM) Analysis
Int. J. Mol. Sci. 2014, 15(11), 19898-19923; doi:10.3390/ijms151119898
Received: 13 August 2014 / Revised: 11 October 2014 / Accepted: 21 October 2014 / Published: 31 October 2014
Cited by 2 | PDF Full-text (875 KB) | HTML Full-text | XML Full-text | Supplementary Files
Abstract
Genetic mutations must be avoided during the production and use of seeds. In the European Union (EU), Directive 2001/18/EC requires any DNA construct introduced via transformation to be stable. Establishing genetic stability is critical for the approval of genetically modified organisms (GMOs). In
[...] Read more.
Genetic mutations must be avoided during the production and use of seeds. In the European Union (EU), Directive 2001/18/EC requires any DNA construct introduced via transformation to be stable. Establishing genetic stability is critical for the approval of genetically modified organisms (GMOs). In this study, genetic stability of two GMOs was examined using high resolution melting (HRM) analysis and real-time polymerase chain reaction (PCR) employing Scorpion primers for amplification. The genetic variability of the transgenic insert and that of the flanking regions in a single oilseed rape variety (GT73) and a stacked maize (MON88017 × MON810) was studied. The GT73 and the 5' region of MON810 showed no instabilities in the examined regions. However; two out of 100 analyzed samples carried a heterozygous point mutation in the 3' region of MON810 in the stacked variety. These results were verified by direct sequencing of the amplified PCR products as well as by sequencing of cloned PCR fragments. The occurrence of the mutation suggests that the 5' region is more suitable than the 3' region for the quantification of MON810. The identification of the single nucleotide polymorphism (SNP) in a stacked event is in contrast to the results of earlier studies of the same MON810 region in a single event where no DNA polymorphism was found. Full article
(This article belongs to the Special Issue Detection and Safety Assessment of Genetically Modified Organisms)
Open AccessArticle Recombinase Polymerase Amplification (RPA) of CaMV-35S Promoter and nos Terminator for Rapid Detection of Genetically Modified Crops
Int. J. Mol. Sci. 2014, 15(10), 18197-18205; doi:10.3390/ijms151018197
Received: 14 August 2014 / Revised: 20 September 2014 / Accepted: 29 September 2014 / Published: 10 October 2014
Cited by 4 | PDF Full-text (1448 KB) | HTML Full-text | XML Full-text
Abstract
Recombinase polymerase amplification (RPA) is a novel isothermal DNA amplification and detection technology that enables the amplification of DNA within 30 min at a constant temperature of 37–42 °C by simulating in vivo DNA recombination. In this study, based on the regulatory sequence
[...] Read more.
Recombinase polymerase amplification (RPA) is a novel isothermal DNA amplification and detection technology that enables the amplification of DNA within 30 min at a constant temperature of 37–42 °C by simulating in vivo DNA recombination. In this study, based on the regulatory sequence of the cauliflower mosaic virus 35S (CaMV-35S) promoter and the Agrobacterium tumefaciens nopaline synthase gene (nos) terminator, which are widely incorporated in genetically modified (GM) crops, we designed two sets of RPA primers and established a real-time RPA detection method for GM crop screening and detection. This method could reliably detect as few as 100 copies of the target molecule in a sample within 15–25 min. Furthermore, the real-time RPA detection method was successfully used to amplify and detect DNA from samples of four major GM crops (maize, rice, cotton, and soybean). With this novel amplification method, the test time was significantly shortened and the reaction process was simplified; thus, this method represents an effective approach to the rapid detection of GM crops. Full article
(This article belongs to the Special Issue Detection and Safety Assessment of Genetically Modified Organisms)
Open AccessArticle Genetically Modified Flax Expressing NAP-SsGT1 Transgene: Examination of Anti-Inflammatory Action
Int. J. Mol. Sci. 2014, 15(9), 16741-16759; doi:10.3390/ijms150916741
Received: 14 July 2014 / Revised: 2 September 2014 / Accepted: 9 September 2014 / Published: 22 September 2014
PDF Full-text (5058 KB) | HTML Full-text | XML Full-text
Abstract
The aim of the work was to define the influence of dietary supplementation with GM (genetically modified) GT#4 flaxseed cake enriched in polyphenols on inflammation development in mice liver. Mice were given ad libitum isoprotein diets: (1) standard diet; (2) high-fat diet rich
[...] Read more.
The aim of the work was to define the influence of dietary supplementation with GM (genetically modified) GT#4 flaxseed cake enriched in polyphenols on inflammation development in mice liver. Mice were given ad libitum isoprotein diets: (1) standard diet; (2) high-fat diet rich in lard, high-fat diet enriched with 30% of (3) isogenic flax Linola seed cake; and (4) GM GT#4 flaxseed cake; for 96 days. Administration of transgenic and isogenic seed cake lowered body weight gain, of transgenic to the standard diet level. Serum total antioxidant status was statistically significantly improved in GT#4 flaxseed cake group and did not differ from Linola. Serum thiobarbituric acid reactive substances, lipid profile and the liver concentration of pro-inflammatory cytokine tumor necrosis factor-α were ameliorated by GM and isogenic flaxseed cake consumption. The level of pro-inflammatory cytokine interferon-γ did not differ between mice obtaining GM GT#4 and non-GM flaxseed cakes. The C-reactive protein concentration was reduced in animals fed GT#4 flaxseed cake and did not differ from those fed non-GM flaxseed cake-based diet. Similarly, the liver structure of mice consuming diets enriched in flaxseed cake was improved. Dietetic enrichment with GM GT#4 and non-GM flaxseed cakes may be a promising solution for health problems resulting from improper diet. Full article
(This article belongs to the Special Issue Detection and Safety Assessment of Genetically Modified Organisms)
Figures

Open AccessArticle Compositional and Proteomic Analyses of Genetically Modified Broccoli (Brassica oleracea var. italica) Harboring an Agrobacterial Gene
Int. J. Mol. Sci. 2014, 15(9), 15188-15209; doi:10.3390/ijms150915188
Received: 17 June 2014 / Revised: 19 August 2014 / Accepted: 25 August 2014 / Published: 28 August 2014
Cited by 2 | PDF Full-text (1909 KB) | HTML Full-text | XML Full-text | Supplementary Files
Abstract
Previously, we showed improved shelf life for agrobacterial isopentenyltransferase (ipt) transgenic broccoli (Brassica oleracea var. italica), with yield comparable to commercial varieties, because of the protection mechanism offered by molecular chaperones and stress-related proteins. Here, we used proximate analysis
[...] Read more.
Previously, we showed improved shelf life for agrobacterial isopentenyltransferase (ipt) transgenic broccoli (Brassica oleracea var. italica), with yield comparable to commercial varieties, because of the protection mechanism offered by molecular chaperones and stress-related proteins. Here, we used proximate analysis to examine macronutrients, chemical and mineral constituents as well as anti-nutrient and protein changes of ipt-transgenic broccoli and corresponding controls. We also preliminarily assessed safety in mice. Most aspects were comparable between ipt-transgenic broccoli and controls, except for a significant increase in carbohydrate level and a decrease in magnesium content in ipt-transgenic lines 101, 102 and 103, as compared with non-transgenic controls. In addition, the anti-nutrient glucosinolate content was increased and crude fat content decreased in inbred control 104 and transgenic lines as compared with the parental control, “Green King”. Gel-based proteomics detected more than 50 protein spots specifically found in ipt-transgenic broccoli at harvest and after cooking; one-third of these proteins showed homology to potential allergens that also play an important role in plant defense against stresses and senescence. Mice fed levels of ipt-transgenic broccoli mimicking the 120 g/day of broccoli eaten by a 60-kg human adult showed normal growth and immune function. In conclusion, the compositional and proteomic changes attributed to the transgenic ipt gene did not affect the growth and immune response of mice under the feeding regimes examined. Full article
(This article belongs to the Special Issue Detection and Safety Assessment of Genetically Modified Organisms)
Open AccessArticle Development and Application of Loop-Mediated Isothermal Amplification Assays for Rapid Visual Detection of cry2Ab and cry3A Genes in Genetically-Modified Crops
Int. J. Mol. Sci. 2014, 15(9), 15109-15121; doi:10.3390/ijms150915109
Received: 11 July 2014 / Revised: 15 August 2014 / Accepted: 18 August 2014 / Published: 27 August 2014
Cited by 10 | PDF Full-text (1108 KB) | HTML Full-text | XML Full-text | Supplementary Files
Abstract
The cry2Ab and cry3A genes are two of the most important insect-resistant exogenous genes and had been widely used in genetically-modified crops. To develop more effective alternatives for the quick identification of genetically-modified organisms (GMOs) containing these genes, a rapid and visual loop-mediated
[...] Read more.
The cry2Ab and cry3A genes are two of the most important insect-resistant exogenous genes and had been widely used in genetically-modified crops. To develop more effective alternatives for the quick identification of genetically-modified organisms (GMOs) containing these genes, a rapid and visual loop-mediated isothermal amplification (LAMP) method to detect the cry2Ab and cry3A genes is described in this study. The LAMP assay can be finished within 60 min at an isothermal condition of 63 °C. The derived LAMP products can be obtained by a real-time turbidimeter via monitoring the white turbidity or directly observed by the naked eye through adding SYBR Green I dye. The specificity of the LAMP assay was determined by analyzing thirteen insect-resistant genetically-modified (GM) crop events with different Bt genes. Furthermore, the sensitivity of the LAMP assay was evaluated by diluting the template genomic DNA. Results showed that the limit of detection of the established LAMP assays was approximately five copies of haploid genomic DNA, about five-fold greater than that of conventional PCR assays. All of the results indicated that this established rapid and visual LAMP assay was quick, accurate and cost effective, with high specificity and sensitivity. In addition, this method does not need specific expensive instruments or facilities, which can provide a simpler and quicker approach to detecting the cry2Ab and cry3A genes in GM crops, especially for on-site, large-scale test purposes in the field. Full article
(This article belongs to the Special Issue Detection and Safety Assessment of Genetically Modified Organisms)

Review

Jump to: Research

Open AccessReview Characterization and Study of Transgenic Cultivars by Capillary and Microchip Electrophoresis
Int. J. Mol. Sci. 2014, 15(12), 23851-23877; doi:10.3390/ijms151223851
Received: 21 October 2014 / Revised: 15 December 2014 / Accepted: 16 December 2014 / Published: 22 December 2014
Cited by 5 | PDF Full-text (2698 KB) | HTML Full-text | XML Full-text
Abstract
Advances in biotechnology have increased the demand for suitable analytical techniques for the analysis of genetically modified organisms. Study of the substantial equivalence, discrimination between transgenic and non-transgenic cultivars, study of the unintended effects caused by a genetic modification or their response to
[...] Read more.
Advances in biotechnology have increased the demand for suitable analytical techniques for the analysis of genetically modified organisms. Study of the substantial equivalence, discrimination between transgenic and non-transgenic cultivars, study of the unintended effects caused by a genetic modification or their response to diverse situations or stress conditions (e.g., environmental, climatic, infections) are some of the concerns that need to be addressed. Capillary electrophoresis (CE) is emerging as an alternative to conventional techniques for the study and characterization of genetically modified organisms. This article reviews the most recent applications of CE for the analysis and characterization of transgenic cultivars in the last five years. Different strategies have been described depending on the level analyzed (DNA, proteins or metabolites). Capillary gel electrophoresis (CGE) has shown to be particularly useful for the analysis of DNA fragments amplified by PCR. Metabolites and proteins have been mainly separated using capillary zone electrophoresis (CZE) using UV and MS detection. Electrophoretic chips have also proven their ability in the analysis of transgenic cultivars and a section describing the new applications is also included. Full article
(This article belongs to the Special Issue Detection and Safety Assessment of Genetically Modified Organisms)
Open AccessReview Metabolomics of Genetically Modified Crops
Int. J. Mol. Sci. 2014, 15(10), 18941-18966; doi:10.3390/ijms151018941
Received: 18 August 2014 / Revised: 8 October 2014 / Accepted: 9 October 2014 / Published: 20 October 2014
Cited by 12 | PDF Full-text (2426 KB) | HTML Full-text | XML Full-text
Abstract
Metabolomic-based approaches are increasingly applied to analyse genetically modified organisms (GMOs) making it possible to obtain broader and deeper information on the composition of GMOs compared to that obtained from traditional analytical approaches. The combination in metabolomics of advanced analytical methods and bioinformatics
[...] Read more.
Metabolomic-based approaches are increasingly applied to analyse genetically modified organisms (GMOs) making it possible to obtain broader and deeper information on the composition of GMOs compared to that obtained from traditional analytical approaches. The combination in metabolomics of advanced analytical methods and bioinformatics tools provides wide chemical compositional data that contributes to corroborate (or not) the substantial equivalence and occurrence of unintended changes resulting from genetic transformation. This review provides insight into recent progress in metabolomics studies on transgenic crops focusing mainly in papers published in the last decade. Full article
(This article belongs to the Special Issue Detection and Safety Assessment of Genetically Modified Organisms)

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