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Related Concept Videos

Genetic Screens02:46

Genetic Screens

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Genetic screens are tools used to identify genes and mutations responsible for phenotypes of interest. Genetic screens help identify individuals or a group of people at risk of developing  genetic diseases and help them with early intervention, targeted therapy, and reproductive options.
Forward genetic screens
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To learn more about the function of a gene, researchers can observe what happens when the gene is inactivated or “knocked out,” by creating genetically engineered knockout animals. Knockout mice have been particularly useful as models for human diseases such as cancer, Parkinson’s disease, and diabetes.
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Updated: Nov 19, 2025

Reverse Genetics to Engineer Positive-Sense RNA Virus Variants
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Reverse Genetics to Engineer Positive-Sense RNA Virus Variants

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Engineering SARS-CoV-2 using a reverse genetic system.

Xuping Xie1, Kumari G Lokugamage2, Xianwen Zhang1

  • 1Department of Biochemistry and Molecular Biology, University of Texas Medical Branch, Galveston, TX, USA.

Nature Protocols
|January 30, 2021
PubMed
Summary
This summary is machine-generated.

Researchers developed a reverse genetic system for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) to engineer viruses for COVID-19 research. This system facilitates studying the virus and developing countermeasures, vaccines, and diagnostics.

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Area of Science:

  • Virology
  • Molecular Biology
  • Genetics

Background:

  • Reverse genetic systems are essential for virus research and countermeasure development.
  • The COVID-19 pandemic highlighted the need for tools to study SARS-CoV-2.
  • Engineering SARS-CoV-2 presents challenges due to its large genome and toxic elements.

Purpose of the Study:

  • To describe a detailed protocol for engineering recombinant SARS-CoV-2 using a reverse genetic system.
  • To enable researchers to utilize SARS-CoV-2 reverse genetics for basic and translational research.

Main Methods:

  • Development of an infectious complementary DNA (cDNA) clone for SARS-CoV-2.
  • A six-step protocol involving plasmid preparation, DNA fragment assembly, in vitro transcription, electroporation, and virus characterization.
  • Utilizing sophisticated methods to overcome challenges associated with the large SARS-CoV-2 genome.

Main Results:

  • Successful engineering of recombinant SARS-CoV-2.
  • A reproducible protocol enabling researchers to master SARS-CoV-2 reverse genetics.
  • Facilitation of rapid virus engineering for various research applications.

Conclusions:

  • The described reverse genetic system and protocol are crucial for advancing COVID-19 research.
  • This system accelerates the study of SARS-CoV-2, including in vitro and in vivo analyses.
  • The protocol supports the development of vaccines, diagnostics, and antiviral screening methods.