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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

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...
CRISPR01:59

CRISPR

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 Short...
CRISPR and crRNAs02:53

CRISPR and crRNAs

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...
Homologous Recombination02:31

Homologous Recombination

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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Related Experiment Video

Updated: Jun 6, 2026

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms
09:51

Enhanced Genome Editing with Cas9 Ribonucleoprotein in Diverse Cells and Organisms

Published on: May 25, 2018

Fully Modified SpyCas9 Guide RNAs Enable Robust Genome Editing In Cells and In Vivo.

Kim Anh Vu1, Han Zhang1, Nadia Amrani1,2

  • 1RNA Therapeutics Institute, University of Massachusetts Chan Medical School, Worcester, MA 01605, USA.

Biorxiv : the Preprint Server for Biology
|June 5, 2026
PubMed
Summary
This summary is machine-generated.

Chemically stabilized guide RNAs (gRNAs) enhance CRISPR/Cas9 genome editing. This study optimized modified gRNAs for improved in vivo editing efficacy and delivery, overcoming previous activity limitations.

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Last Updated: Jun 6, 2026

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Published on: May 25, 2018

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11:35

Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells

Published on: June 16, 2017

Area of Science:

  • Molecular Biology
  • Biotechnology
  • Genetic Engineering

Background:

  • CRISPR/Cas9 technology is crucial for genome editing.
  • Chemically stabilized guide RNAs (gRNAs) offer potential for improved in vivo editing and delivery.
  • Previous fully modified gRNAs showed reduced Cas9 activity.

Purpose of the Study:

  • To optimize chemically stabilized guide RNAs (gRNAs) for CRISPR/Cas9.
  • To enhance in vivo genome editing efficacy and delivery flexibility.
  • To overcome reduced Cas9 activity associated with modified gRNAs.

Main Methods:

  • Iterative, structure-guided optimization of SpyCas9 gRNAs.
  • Systematic introduction of chemical modifications at each nucleotide position.
  • Incorporation of 2'-amino-RNA, 4'-thio-RNA, and extended nucleic acid (exNA).

Main Results:

  • Developed gRNA designs with 90-100% sugar- or backbone-modified nucleotides.
  • Established modified gRNAs that maintain or enhance editing efficacy in vitro and in vivo.
  • Observed sequence-dependent variability in modification patterns.

Conclusions:

  • Heavily and fully modified gRNAs show promise for CRISPR/Cas9 applications.
  • Optimized gRNAs can improve genome editing tools like nuclease and base editing.
  • These modified gRNAs offer potential for advanced therapeutic genome editing.