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CRISPR/Cas9 Genome Editing

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...

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Updated: Nov 20, 2025

Pooled CRISPR-Based Genetic Screens in Mammalian Cells

Pooled CRISPR-Based Genetic Screens in Mammalian Cells

Published on: September 4, 2019

Extensible combinatorial CRISPR screening in mammalian cells.

Peng Zhou¹, Yuk Kei Wan¹, Becky K C Chan¹

¹Laboratory of Combinatorial Genetics and Synthetic Biology, School of Biomedical Sciences, The University of Hong Kong, Pokfulam, Hong Kong SAR, China.

|January 25, 2021

Summary

This summary is machine-generated.

This study introduces Combinatorial Genetics En Masse (CombiGEM) for analyzing complex genetic interactions using CRISPR screens. It details methods for library construction, screening, and analysis to uncover therapeutic targets.

Keywords:

CRISPR High Throughput Screening

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Last Updated: Nov 20, 2025

Pooled CRISPR-Based Genetic Screens in Mammalian Cells

Pooled CRISPR-Based Genetic Screens in Mammalian Cells

Published on: September 4, 2019

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Genome Editing in Mammalian Cell Lines using CRISPR-Cas

Published on: April 11, 2019

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

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

Published on: June 16, 2017

Area of Science:

Molecular Biology
Genetics
Bioinformatics

Background:

Understanding high-order genetic perturbations is crucial for deciphering biological processes.
Identifying combinations of therapeutic targets requires advanced genetic screening methods.
Existing methods may not efficiently analyze complex genetic interactions.

Purpose of the Study:

To present a detailed protocol for combinatorial CRISPR screening in mammalian cells using CombiGEM.
To describe the construction of multiplexed guide RNA libraries for high-throughput screening.
To introduce CombiPIPE, an analytical pipeline for mapping genetic interactions.

Main Methods:

Utilized the CRISPR-Cas system combined with Combinatorial Genetics En Masse (CombiGEM).
Developed methods for building multiplexed guide RNA libraries.
Performed combinatorial CRISPR screens in mammalian cells and analyzed data with CombiPIPE.

Main Results:

Successfully established a protocol for combinatorial CRISPR screening.
Demonstrated the capability of CombiGEM for analyzing high-order genetic perturbations.
CombiPIPE effectively maps two- and three-way genetic interactions.

Conclusions:

CombiGEM provides a powerful platform for systematic analysis of complex genetic interactions.
The presented protocol and analytical pipeline facilitate the discovery of therapeutic target combinations.
This approach enhances our understanding of biological systems through high-order genetic perturbation analysis.