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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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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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Gene Digital Circuits Based on CRISPR-Cas Systems and Anti-CRISPR Proteins
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Genome dependent Cas9/gRNA search time underlies sequence dependent gRNA activity.

E A Moreb1, M D Lynch2

  • 1Department of Biomedical Engineering, Duke University, Durham, USA.

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Guide CRISPR-Cas9 gene editing with insights into guide RNA (gRNA) efficiency. This study reveals that the Cas9/gRNA complex

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR-Cas9 is a versatile gene editing technology.
  • Guide RNA (gRNA) sequence influences Cas9 cleavage efficiency.
  • Existing prediction algorithms show limitations across datasets and species.

Purpose of the Study:

  • To investigate sequence features impacting gRNA activity.
  • To understand discrepancies in current gRNA prediction models.
  • To identify factors driving inter-species differences in gRNA efficacy.

Main Methods:

  • Retrospective analysis of 44 published datasets.
  • Examination of sequence features affecting gRNA activity.
  • Comparative analysis across different species.

Main Results:

  • gRNA activity is primarily determined by target site accessibility, not on-target activity.
  • This accessibility influences sequence-dependent differences in gRNA efficacy.
  • Inter-species variations in gRNA activity are linked to target site finding.

Conclusions:

  • Cas9/gRNA complex's ability to locate target DNA is crucial for activity.
  • Understanding target site recognition improves gRNA design and Cas9 variant development.
  • This research provides a foundation for optimizing CRISPR-Cas9 applications.