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Updated: Feb 1, 2026
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Selection-dependent and Independent Generation of CRISPR/Cas9-mediated Gene Knockouts in Mammalian Cells
Published on: June 16, 2017
13.3K
CRISPR/Cas9-Based Chemogenomic Profiling in Mammalian Cells.
Dominic Hoepfner1, Gregory McAllister2, Gregory R Hoffman2
1Novartis Institutes for BioMedical Research, Novartis Pharma AG, Basel, Switzerland. dominic.hoepfner@novartis.com.
Methods in Molecular Biology (Clifton, N.J.)
|December 7, 2018
Summary
Chemogenomic profiling identifies drug targets by analyzing gene modifications. This study details a CRISPR/Cas9 protocol for precise, genome-wide profiling in mammalian cells, overcoming previous limitations.
Area of Science:
- Pharmacology
- Genomics
- Molecular Biology
Background:
- Chemogenomic profiling identifies drug targets and mechanisms of action.
- Genome-wide gene modulation studies are essential but historically limited to fungi.
- Previous mammalian cell approaches using RNAi suffered from off-target effects and noise.
Purpose of the Study:
- To present a detailed protocol for chemogenomic profiling using the CRISPR/Cas9 system.
- To enable precise, genome-wide target identification of chemical probes in mammalian cells.
Main Methods:
- Leveraging the CRISPR/Cas9 system for precise DNA editing at defined loci.
- Implementing genome-wide CRISPR/Cas9 screening for chemogenomic profiling.
- Developing a protocol for high-resolution analysis of gene modulation effects.
Main Results:
- Demonstrated the feasibility of CRISPR/Cas9 for genome-wide chemogenomic profiling.
- Enabled precise identification of cellular hypersensitivity and resistance patterns.
- Overcame limitations of previous methods in mammalian systems.
Conclusions:
- CRISPR/Cas9 system provides a powerful tool for unbiased chemogenomic profiling.
- This protocol facilitates accurate target identification for diverse chemical probes.
- Advances high-resolution genetic screening in mammalian cells for drug discovery.
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