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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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CRISPR and crRNAs02:53

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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.
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Related Experiment Video

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Expanding CRISPR/Cas9 Genome Editing Capacity in Zebrafish Using SaCas9.

Yan Feng1, Cheng Chen1, Yuxiang Han1

  • 1Laboratory of Chemical Genomics, School of Chemical Biology and Biotechnology, Peking University Shenzhen Graduate School, 518055, China.

G3 (Bethesda, Md.)
|June 19, 2016
PubMed
Summary

Researchers expanded CRISPR-Cas9 genome editing in zebrafish by using Staphylococcus aureus Cas9 (SaCas9). This broadens the available target sites, enhancing zebrafish

Keywords:
CRISPR/Cas9KKH SaCas9 variantSaCas9gene editingzebrafish

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Area of Science:

  • Molecular Biology
  • Genetics
  • Zebrafish Models

Background:

  • The CRISPR-Cas9 system is a powerful tool for genome editing in zebrafish.
  • The commonly used Streptococcus pyogenes Cas9 (SpCas9) has limitations due to its strict protospacer-adjacent motif (PAM) requirement (5'-NGG-3'), restricting targetable sequences.

Purpose of the Study:

  • To investigate the efficacy of alternative Cas9 orthologs for genome editing in zebrafish.
  • To expand the range of targetable sequences for CRISPR-Cas9 in the zebrafish genome.
  • To enhance the utility of zebrafish as a model organism for genetic studies.

Main Methods:

  • Utilized Cas9 orthologs from Staphylococcus aureus (SaCas9) and its KKH variant for targeted mutagenesis in zebrafish.
  • Confirmed the activity of a previously reported SpCas9 variant (VQR) in zebrafish.
  • Performed bioinformatics analysis to assess the expanded target site repertoire.

Main Results:

  • SaCas9 and its KKH variant successfully induced targeted mutagenesis with high frequency in zebrafish.
  • The SpCas9 VQR variant also induced targeted mutations, consistent with prior findings.
  • Bioinformatics analysis indicated a significant expansion of available Cas9 target sites in the zebrafish genome.

Conclusions:

  • Staphylococcus aureus Cas9 (SaCas9) and its variants are effective tools for zebrafish genome editing.
  • The expanded target repertoire significantly increases the versatility of CRISPR-Cas9 in zebrafish.
  • These advancements will further facilitate genetic studies in zebrafish, advancing vertebrate biology research.