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CRISPR/Cas9 screening using unique molecular identifiers.

ABSTRACT: Loss-of-function screening by CRISPR/Cas9 gene knockout with pooled, lentiviral guide libraries is a widely applicable method for systematic identification of genes contributing to diverse cellular phenotypes. Here, Random Sequence Labels (RSLs) are incorporated into the guide library, which act as unique molecular identifiers (UMIs) to allow massively parallel lineage tracing and lineage dropout screening. RSLs greatly improve the reproducibility of results by increasing both the precision and the accuracy of screens. They reduce the number of cells needed to reach a set statistical power, or allow a more robust screen using the same number of cells.

SUBMITTER: Schmierer B

PROVIDER: S-EPMC5658704 | biostudies-literature | 2017 Oct

REPOSITORIES: biostudies-literature

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CRISPR/Cas9 screening using unique molecular identifiers.

Schmierer Bernhard B Botla Sandeep K SK Zhang Jilin J Turunen Mikko M Kivioja Teemu T Taipale Jussi J

Molecular systems biology 20171009 10

Loss-of-function screening by CRISPR/Cas9 gene knockout with pooled, lentiviral guide libraries is a widely applicable method for systematic identification of genes contributing to diverse cellular phenotypes. Here, Random Sequence Labels (RSLs) are incorporated into the guide library, which act as unique molecular identifiers (UMIs) to allow massively parallel lineage tracing and lineage dropout screening. RSLs greatly improve the reproducibility of results by increasing both the precision and ...[more]

PMID: 28993443

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