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Generating Custom Pooled CRISPR Libraries for Genetic Dissection of Biological Pathways

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Membrane Trafficking

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2473))

Abstract

Genetic screens are a classic approach to dissecting biological pathways including membrane trafficking. Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 have enabled the utility of this approach in diploid models, including cultured mammalian cells. Here, we present detailed protocols for generating custom CRISPR libraries. These methods are useful for generating genome-wide libraries for new model organisms that lack an existing genome-wide library, and for generating smaller focused libraries.

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Acknowledgments

This work was supported by a NIH fellowship award F32AG062039 (to D.G.).

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Correspondence to Daniel R. Gulbranson .

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Gulbranson, D.R. (2022). Generating Custom Pooled CRISPR Libraries for Genetic Dissection of Biological Pathways. In: Shen, J. (eds) Membrane Trafficking. Methods in Molecular Biology, vol 2473. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2209-4_21

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  • DOI: https://doi.org/10.1007/978-1-0716-2209-4_21

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  • Publisher Name: Humana, New York, NY

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  • Online ISBN: 978-1-0716-2209-4

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