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Mol Syst Biol. 2017 Oct 9;13(10):945. doi: 10.15252/msb.20177834.

CRISPR/Cas9 screening using unique molecular identifiers.

Author information

1
Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden.
2
Genome-Scale Biology Research Program, Faculty of Medicine, University of Helsinki, Helsinki, Finland.
3
Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Stockholm, Sweden jussi.taipale@ki.se.

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.

KEYWORDS:

CRISPR/Cas; genetic screening; massively parallel lineage tracing; unique molecular identifiers

PMID:
28993443
PMCID:
PMC5658704
DOI:
10.15252/msb.20177834
[Indexed for MEDLINE]
Free PMC Article

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