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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

208
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...
208
CRISPR01:59

CRISPR

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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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Homologous Recombination02:31

Homologous Recombination

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The basic reaction of homologous recombination (HR) involves two chromatids that contain DNA sequences sharing a significant stretch of identity. One of these sequences uses a strand from another as a template to synthesize DNA in an enzyme-catalyzed reaction. The final product is a novel amalgamation of the two substrates. To ensure an accurate recombination of sequences, HR is restricted to the S and G2 phases of the cell cycle. At these stages, the DNA has been replicated already and the...
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Conservative Site-specific Recombination and Phase Variation02:53

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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.
The recognition sites for Cre recombinase called LoxP...
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CRISPR and crRNAs02:53

CRISPR and crRNAs

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Bacteria and archaea are susceptible to viral infections just like eukaryotes; therefore, they have developed a unique adaptive immune system to protect themselves. Clustered regularly interspaced short palindromic repeats and CRISPR-associated proteins (CRISPR-Cas) are present in more than 45% of known bacteria and 90% of known archaea.
The CRISPR-Cas system stores a copy of foreign DNA in the host genome and uses it to identify the foreign DNA upon reinfection. CRISPR-Cas has three different...
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Updated: Sep 8, 2025

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
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Tools for Efficient Genome Editing; ZFN, TALEN, and CRISPR.

Yasaman Shamshirgaran1, Jun Liu2, Huseyin Sumer3

  • 1Laboratory of Clinical Chemistry, Sahlgrenska University Hospital, Gothenburg, Sweden.

Methods in Molecular Biology (Clifton, N.J.)
|June 13, 2022
PubMed
Summary

Genome editing tools like CRISPR-Cas have advanced significantly. Recent findings suggest Cas9-induced mutations are non-random, impacting gene function studies.

Keywords:
Cas9DNA repairGenome editingTALENZFN

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • The last two decades have seen major advances in genome editing technologies.
  • Programmable nucleases, including ZFNs, TALENs, and CRISPR-Cas systems, introduce targeted DNA double-strand breaks (DSBs) in eukaryotic cells.
  • Cellular DNA repair pathways are utilized to create insertions and deletions (indels) at DSBs for gene function studies.

Purpose of the Study:

  • To provide a concise overview of genome editing tools.
  • To describe the DNA repair pathways involved in genome editing outcomes.
  • To highlight recent findings on the nature of Cas9-induced mutations.

Main Methods:

  • Review of genome editing technologies (ZFNs, TALENs, CRISPR-Cas).
  • Analysis of DNA repair pathways (non-homologous end joining, homology-directed repair).
  • Discussion of computational tools for analyzing Cas9-induced mutations.

Main Results:

  • Genome editing tools have revolutionized life sciences research.
  • Recent computational analyses suggest Cas9-induced mutations exhibit non-random patterns.
  • Newer CRISPR-centric tools like base and prime editors expand the genome editing toolbox.

Conclusions:

  • Genome editing technologies offer powerful capabilities for biological research.
  • Understanding the DNA repair mechanisms underlying indels is crucial.
  • The non-random nature of Cas9-induced mutations necessitates refined analytical approaches.