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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

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.
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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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Related Experiment Video

Updated: Mar 16, 2026

Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution
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Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution

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Methods for Optimizing CRISPR-Cas9 Genome Editing Specificity.

Josh Tycko1, Vic E Myer1, Patrick D Hsu2

  • 1Editas Medicine, 300 Third Street, Cambridge, MA 02142, USA.

Molecular Cell
|August 6, 2016
PubMed
Summary
This summary is machine-generated.

CRISPR-Cas9 gene editing advances enhance understanding of gene function and offer therapeutic potential for genetic disorders. Ongoing research focuses on improving Cas9 specificity to minimize off-target effects for safer genome engineering applications.

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A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
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Genome Editing in Mammalian Cell Lines using CRISPR-Cas
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Using Sniper-Cas9 to Minimize Off-target Effects of CRISPR-Cas9 Without the Loss of On-target Activity Via Directed Evolution
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A Standard Methodology to Examine On-site Mutagenicity As a Function of Point Mutation Repair Catalyzed by CRISPR/Cas9 and SsODN in Human Cells
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Genome Editing in Mammalian Cell Lines using CRISPR-Cas
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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR-Cas9 is a powerful genome engineering tool with applications in basic research and therapeutics.
  • Understanding and improving Cas9 specificity is crucial for its safe and effective use.
  • Off-target DNA binding and cleavage by Cas9 are known limitations.

Purpose of the Study:

  • To review challenges and breakthroughs in CRISPR-Cas9 specificity.
  • To provide a practical guide for optimizing genome editing specificity.
  • To highlight strategies for enhancing Cas9 precision and sensitivity.

Main Methods:

  • Review of current literature on CRISPR-Cas9 specificity strategies.
  • Analysis of advancements in guide RNA selection and engineering.
  • Evaluation of novel Cas9 enzymes and off-target detection methods.

Main Results:

  • Significant progress has been made in improving Cas9 specificity through various approaches.
  • Guide RNA optimization, protein engineering, and novel enzymes enhance precision.
  • Advanced detection methods aid in identifying and mitigating off-target effects.

Conclusions:

  • CRISPR-Cas9 technology is rapidly advancing in specificity, offering greater precision for gene function studies and therapeutic applications.
  • Standardization of off-target activity measurement and reporting is needed.
  • Continued vigilance and improvement in specificity are essential for the responsible development of genome editing technologies.