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Related Concept Videos

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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CRISPR01:59

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CRISPR/Cas9 Genome Editing01:28

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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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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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The Antiviral System of Bacteria and Archaea: CRISPR01:23

The Antiviral System of Bacteria and Archaea: CRISPR

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CRISPR stands for Clustered Regularly Interspaced Short Palindromic Repeats is a adaptive immune system found in bacteria and archaea that protects against viral infections. This system enables prokaryotic cells to identify, remember, and neutralize foreign genetic elements, primarily bacteriophages, by storing fragments of the invader’s DNA as a genetic memory.The CRISPR immune response begins during an initial infection. Cas (CRISPR-associated) proteins play a central role in this...
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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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CIRCLE-Seq for Interrogation of Off-Target Gene Editing
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Keeping CRISPR/Cas on-Target.

Muhammad Jamal1, Faheem Ahmed Khan, Lin Da

  • 1State Key Laboratory of Agriculture Microbiology, Huazhong Agricultural University, Wuhan, Hubei, 430070, P. R. China.

Current Issues in Molecular Biology
|October 11, 2015
PubMed
Summary
This summary is machine-generated.

CRISPR/Cas gene editing offers high efficiency but faces off-target mutations. This review details strategies for enhancing CRISPR specificity through guide RNA design and Cas9 modification for safer genome engineering.

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • CRISPR/Cas is a powerful genome editing tool derived from microbial immunity.
  • Current CRISPR/Cas systems exhibit high efficiency but suffer from off-target mutations, limiting their application.
  • Addressing specificity challenges is crucial for advancing CRISPR-mediated gene editing.

Purpose of the Study:

  • To review recent advancements in improving CRISPR/Cas specificity.
  • To discuss methods for rational guide RNA (gRNA) design and Cas9 endonuclease modification.
  • To explore techniques for screening and evaluating off-target mutations.

Main Methods:

  • Focus on rational design principles for gRNA.
  • Investigate modifications to the Cas9 endonuclease to enhance specificity.
  • Summarize various screening assays for detecting off-target mutations.

Main Results:

  • Rational gRNA design and Cas9 modifications effectively reduce off-target mutations.
  • Specific screening methods aid in identifying and quantifying unintended edits.
  • Combined strategies significantly improve the precision of CRISPR/Cas gene editing.

Conclusions:

  • Enhancing CRISPR specificity is achievable through targeted gRNA design and Cas9 engineering.
  • Effective screening assays are essential for validating the safety and efficacy of gene editing.
  • Optimized CRISPR/Cas systems hold great promise for successful genome engineering applications.