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Methods and Protocols, Volume 4, Issue 1

March 2021 - 23 articles

Cover Story: Bombyx mori is a model organism for lepidopteran genetics which is economically important for producing silk and is used in the textile, biomedical, and cosmetics sectors. Silkworms can also act as genetically engineered bioreactors to produce recombinant proteins. CRISPR/Cas9 technology is an efficient method of generating heritable gene-specific DNA modifications. After CRISPR/Cas9 mutagenesis, silkworms must be molecularly screened to identify individuals carrying a suitable DNA modification for selective breeding. We describe an efficient screening workflow involving a heteroduplex assay and Sanger sequencing for the characterization of mutations, and the amplification refractory mutation system PCR for the subsequent detection of the modification, enabling the establishment of B. mori mutant lines stably inheriting specific mutations. View this paper.
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Articles (23)

  • Communication
  • Open Access
3 Citations
2,919 Views
8 Pages

11 February 2021

Human rhinovirus infections are a major trigger for acute exacerbations of lower airway diseases, including asthma and chronic obstructive pulmonary disease. Disease exacerbation is thought to be regulated via double-stranded RNA (dsRNA)-mediated sig...

  • Commentary
  • Open Access
8 Citations
3,532 Views
5 Pages

The Protocol Gap

  • Michael G. Weller

3 February 2021

Although peer review is considered one of the main pillars of modern science, experimental methods and protocols seem to be not a rigorous subject of this process in many papers. Commercial equipment, test kits, labeling kits, previously published co...

  • Review
  • Open Access
26 Citations
5,129 Views
13 Pages

The Past, Present and Future of Flow Cytometry in Central Nervous System Malignancies

  • Evrysthenis Vartholomatos,
  • George Vartholomatos,
  • George A. Alexiou and
  • Georgios S. Markopoulos

26 January 2021

Central nervous system malignancies (CNSMs) are categorized among the most aggressive and deadly types of cancer. The low median survival in patients with CNSMs is partly explained by the objective difficulties of brain surgeries as well as by the ac...

  • Perspective
  • Open Access
262 Citations
18,104 Views
18 Pages

Recent Advances in miRNA Delivery Systems

  • Ishani Dasgupta and
  • Anushila Chatterjee

20 January 2021

MicroRNAs (miRNAs) represent a family of short non-coding regulatory RNA molecules that are produced in a tissue and time-specific manner to orchestrate gene expression post-transcription. MiRNAs hybridize to target mRNA(s) to induce translation repr...

  • Article
  • Open Access
11 Citations
5,058 Views
14 Pages

Validity and Cost-Effectiveness of Pediatric Home Respiratory Polygraphy for the Diagnosis of Obstructive Sleep Apnea in Children: Rationale, Study Design, and Methodology

  • Esther Oceja,
  • Paula Rodríguez,
  • María José Jurado,
  • Maria Luz Alonso,
  • Genoveva del Río,
  • María Ángeles Villar,
  • Olga Mediano,
  • Marian Martínez,
  • Santiago Juarros and
  • Milagros Merino
  • + 5 authors

Obstructive sleep apnea (OSA) in children is a prevalent, albeit largely undiagnosed disease associated with a large spectrum of morbidities. Overnight in-lab polysomnography remains the gold standard diagnostic approach, but is time-consuming, incon...

  • Protocol
  • Open Access
12 Citations
7,271 Views
24 Pages

The expansion and intensification of agriculture have led to global declines in biodiversity. This paper presents a systematic review protocol to clarify under what management and landscape contexts diversified farming practices are effective at impr...

  • Letter
  • Open Access
20 Citations
5,190 Views
11 Pages

Inactivation of Material from SARS-CoV-2-Infected Primary Airway Epithelial Cell Cultures

  • Kaitlyn A. Barrow,
  • Lucille M. Rich,
  • Elizabeth R. Vanderwall,
  • Stephen R. Reeves,
  • Jennifer A. Rathe,
  • Maria P. White and
  • Jason S. Debley

Given that the airway epithelium is the initial site of infection, study of primary human airway epithelial cells (AEC) infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) will be crucial to improved understanding of viral entr...

  • Study Protocol
  • Open Access
3 Citations
3,502 Views
12 Pages

Novel In Vivo Mouse Cryoablation Model to Explore Unique Therapeutic Approaches for Premalignant Columnar Lesions

  • Ana C. P. Correia,
  • Danielle Straub,
  • Silvia Calpe and
  • Kausilia K. Krishnadath

Patients with epithelial metaplasias have an increased risk of developing malignancies. In Barrett’s esophagus, neo-columnar epithelium develops proximal to the squamous-columnar junction (SCJ) in the esophagus as the result of prolonged exposu...

  • Article
  • Open Access
8 Citations
4,495 Views
11 Pages

30 December 2020

Over the past two decades, it has become increasingly evident that microRNAs (miRNA) play a major role in human diseases such as cancer and cardiovascular diseases. Moreover, their easy detection in circulation has made them a tantalizing target for...

  • Protocol
  • Open Access
9 Citations
4,876 Views
14 Pages

An Efficient Workflow for Screening and Stabilizing CRISPR/Cas9-Mediated Mutant Lines in Bombyx mori

  • Daniel Brady,
  • Alessio Saviane,
  • Silvia Cappellozza and
  • Federica Sandrelli

29 December 2020

The domestic silkworm Bombyx mori is extensively studied as a model organism for lepidopteran genetics and has an economic value in silk production. Silkworms also have applications in biomedical and cosmetic industries, and the production of mutant...

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Methods Protoc. - ISSN 2409-9279