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Related Concept Videos

CRISPR/Cas9 Genome Editing01:28

CRISPR/Cas9 Genome Editing

625
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...
625
CRISPR01:59

CRISPR

53.4K
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...
53.4K

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Related Experiment Video

Updated: Oct 14, 2025

Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9
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Generation of Genomic Deletions in Mammalian Cell Lines via CRISPR/Cas9

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Optimized protocol to create deletion in adherent cell lines using CRISPR/Cas9 system.

Umer Farooq1,2, Dimple Notani1

  • 1National Centre for Biological Sciences, Tata Institute for Fundamental Research, Bangalore, Karnataka 560065, India.

STAR Protocols
|November 8, 2021
PubMed
Summary
This summary is machine-generated.

This study provides a detailed CRISPR/Cas9 protocol for creating knockout cell lines. The method is effective for both coding and non-coding regions in common cell lines.

Keywords:
CRISPRGene ExpressionGenomicsMolecular BiologySingle Cell

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Area of Science:

  • Molecular Biology
  • Genetics
  • Biotechnology

Background:

  • The CRISPR/Cas9 system offers versatile genome editing capabilities.
  • Generating knockout cell lines is crucial for functional genomics research.

Purpose of the Study:

  • To present a step-by-step protocol for generating knockout cell lines using the CRISPR/Cas9 system.
  • To detail the application of this protocol for both coding and non-coding genomic regions.

Main Methods:

  • Utilized the CRISPR/Cas9 gene editing tool.
  • Developed a protocol specifically for creating knockout cell lines.
  • Tested the protocol on adherent cell lines, including HeLa and MCF7.

Main Results:

  • Successfully generated knockout cell lines using the described CRISPR/Cas9 protocol.
  • Demonstrated the protocol's applicability to coding and non-coding regions.
  • Confirmed the protocol's effectiveness on HeLa and MCF7 cell lines.

Conclusions:

  • The presented CRISPR/Cas9 protocol is a reliable method for generating knockout cell lines.
  • The protocol is adaptable for various adherent cell lines with minor modifications.
  • This method facilitates functional studies by enabling targeted gene disruption.