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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...
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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: May 14, 2026

Establishment of Genome-edited Human Pluripotent Stem Cell Lines: From Targeting to Isolation
09:51

Establishment of Genome-edited Human Pluripotent Stem Cell Lines: From Targeting to Isolation

Published on: February 2, 2016

RNA-programmed genome editing in human cells.

Martin Jinek1, Alexandra East, Aaron Cheng

  • 1Howard Hughes Medical Institute, University of California, Berkeley , Berkeley , United States ; Department of Molecular and Cell Biology , University of California, Berkeley , Berkeley , United States.

Elife
|February 7, 2013
PubMed
Summary
This summary is machine-generated.

Scientists demonstrate that Cas9 protein and guide RNA can create targeted DNA breaks in human cells, enabling precise genome editing. This RNA-programmed system offers a straightforward method for genetic modification in human cells.

Keywords:
Cas9Humanendonucleasegenome editing

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Related Experiment Videos

Last Updated: May 14, 2026

Establishment of Genome-edited Human Pluripotent Stem Cell Lines: From Targeting to Isolation
09:51

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Published on: February 2, 2016

Introducing Point Mutations into Human Pluripotent Stem Cells Using Seamless Genome Editing
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Published on: May 10, 2020

CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery
07:49

CRISPR Epigenome Editing in Human Cells using Plasmid DNA Transfection and mRNA Nucleofection Delivery

Published on: May 30, 2025

Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • Type II CRISPR systems utilize Cas9 endonuclease for bacterial defense against foreign DNA.
  • Cas9's ability to be programmed by RNA for targeted DNA cleavage is a key feature.

Purpose of the Study:

  • To investigate the functionality of bacterial Cas9 in human cells for genome editing.
  • To determine the requirements and limitations of Cas9-mediated DNA cleavage in a human cellular context.

Main Methods:

  • Transfection of human cells with Cas9 and hybrid guide RNAs.
  • Analysis of Cas9-induced double-strand DNA breaks (DSBs) at specific genomic loci.
  • Biochemical assays using cell extracts to identify limiting factors for Cas9 activity.

Main Results:

  • Cas9 successfully assembles with guide RNAs in human cells.
  • Targeted DSBs are induced at sites complementary to the guide RNA sequence.
  • RNA expression and Cas9 assembly are identified as limiting factors for cleavage.
  • 3' RNA sequence extension enhances in vivo DNA targeting.

Conclusions:

  • RNA-programmed genome editing using Cas9 is feasible in human cells.
  • This technology provides a facile strategy for site-specific genetic modifications.
  • Understanding limiting factors can optimize CRISPR-Cas9 applications in human therapeutics.