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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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The CRISPR-Cas system serves as a bacterial defense mechanism against invading genetic elements such as viruses and plasmids, forming the foundation for its adaptation as a powerful genome-editing tool. Originally discovered in prokaryotes, this system has been repurposed to revolutionize genetic engineering across a wide range of organisms, including plants, animals, and humans. The core component, Cas9, is an endonuclease derived from Streptococcus pyogenes, capable of introducing...
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CRISPR01:59

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Genome editing technologies allow scientists to modify an organism’s DNA via the addition, removal, or rearrangement of genetic material at specific genomic locations. These types of techniques could potentially be used to cure genetic disorders such as hemophilia and sickle cell anemia. One popular and widely used DNA-editing research tool that could lead to safe and effective cures for genetic disorders is the CRISPR-Cas9 system. CRISPR-Cas9 stands for Clustered Regularly Interspaced...
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Because the DNA segments are cut and reorganized in a direction-specific manner, site-specific recombination has emerged as an efficient genetic engineering technique. Flippase and Cyclization recombinases or Flp and Cre, respectively, are two members of the tyrosine recombinase family derived from bacteriophages, that are used to mediate site-specific DNA insertions, deletions, and targeted expression of proteins in mammalian cell lines.
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A New Toolkit for Evaluating Gene Functions using Conditional Cas9 Stabilization
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A versatile, high-efficiency platform for CRISPR-based gene activation.

Amy J Heidersbach1, Kristel M Dorighi2, Javier A Gomez3

  • 1Department of Molecular Biology, Genentech Inc., South San Francisco, CA, USA. heidersbach.amy@gene.com.

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|February 22, 2023
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We developed a new CRISPR-mediated transcriptional activation (CRISPRa) system that efficiently generates CRISPRa-ready cell lines. This platform enhances gene activation and broadens CRISPRa applications in various cellular contexts.

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Area of Science:

  • Gene editing and functional genomics
  • Molecular biology and genetic engineering

Background:

  • CRISPR-mediated transcriptional activation (CRISPRa) is crucial for gene expression studies.
  • Existing methods for creating CRISPRa-competent cell lines have limitations.

Purpose of the Study:

  • To develop a high-efficiency, self-selecting CRISPRa enrichment strategy.
  • To enable rapid production of CRISPRa-ready cell populations.
  • To enhance CRISPRa functionality and expand its utility.

Main Methods:

  • Utilized a self-selecting CRISPRa enrichment strategy combined with piggyBac transposon technology.
  • Developed an optimized guide RNA scaffold for enhanced CRISPRa function.
  • Created a synthetic guide RNA toolset for transient gene activation.

Main Results:

  • Successfully generated CRISPRa-ready cell populations efficiently.
  • Demonstrated significantly enhanced CRISPRa functionality with the optimized scaffold.
  • Enabled transient, population-wide gene activation using the synthetic guide RNA toolset.

Conclusions:

  • The developed platform offers a versatile and efficient approach for CRISPRa applications.
  • This technology overcomes limitations of current CRISPRa cell line generation methods.
  • It significantly enhances the potential of CRISPRa in diverse cellular research settings.