Genome-Wide CRISPR/Cas9 Screening for Identification of Cancer Genes in Cell Lines

Charles H Adelmann^{1

2

3

4}, Tim Wang^{1

2

3

4}, David M Sabatini^{5

6

7

8

9}, Eric S Lander^{10

11

12}

Affiliations

¹ Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA.
² Whitehead Institute for Biomedical Research, Cambridge, MA, USA.
³ Howard Hughes Medical Institute Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA.
⁴ David H. Koch Institute for Integrative Cancer Research at MIT, Cambridge, MA, USA.
⁵ Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA. sabatini@wi.mit.edu.
⁶ Whitehead Institute for Biomedical Research, Cambridge, MA, USA. sabatini@wi.mit.edu.
⁷ Howard Hughes Medical Institute Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA. sabatini@wi.mit.edu.
⁸ David H. Koch Institute for Integrative Cancer Research at MIT, Cambridge, MA, USA. sabatini@wi.mit.edu.
⁹ Broad Institute of MIT and Harvard, Cambridge, MA, USA. sabatini@wi.mit.edu.
¹⁰ Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA. lander@broadinstitute.org.
¹¹ Broad Institute of MIT and Harvard, Cambridge, MA, USA. lander@broadinstitute.org.
¹² Department of Systems Biology, Harvard Medical School, Boston, MA, USA. lander@broadinstitute.org.

PMID: 30542996
DOI: 10.1007/978-1-4939-8967-6_10

Genome-Wide CRISPR/Cas9 Screening for Identification of Cancer Genes in Cell Lines

Charles H Adelmann et al. Methods Mol Biol. 2019.

. 2019:1907:125-136.

doi: 10.1007/978-1-4939-8967-6_10.

Authors

Charles H Adelmann^{1

2

3

4}, Tim Wang^{1

2

3

4}, David M Sabatini^{5

6

7

8

9}, Eric S Lander^{10

11

12}

Affiliations

¹ Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA.
² Whitehead Institute for Biomedical Research, Cambridge, MA, USA.
³ Howard Hughes Medical Institute Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA.
⁴ David H. Koch Institute for Integrative Cancer Research at MIT, Cambridge, MA, USA.
⁵ Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA. sabatini@wi.mit.edu.
⁶ Whitehead Institute for Biomedical Research, Cambridge, MA, USA. sabatini@wi.mit.edu.
⁷ Howard Hughes Medical Institute Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA. sabatini@wi.mit.edu.
⁸ David H. Koch Institute for Integrative Cancer Research at MIT, Cambridge, MA, USA. sabatini@wi.mit.edu.
⁹ Broad Institute of MIT and Harvard, Cambridge, MA, USA. sabatini@wi.mit.edu.
¹⁰ Department of Biology, Massachusetts Institute of Technology, Cambridge, MA, USA. lander@broadinstitute.org.
¹¹ Broad Institute of MIT and Harvard, Cambridge, MA, USA. lander@broadinstitute.org.
¹² Department of Systems Biology, Harvard Medical School, Boston, MA, USA. lander@broadinstitute.org.

PMID: 30542996
DOI: 10.1007/978-1-4939-8967-6_10

Abstract

In this protocol, pooled sgRNA libraries targeting thousands of genes are computationally designed, generated using microarray-based synthesis techniques, and packaged into lentiviral particles. Target cells of interest are transduced with the lentiviral sgRNA pools to generate a collection of knockout mutants-via Cas9-mediated genomic cleavage-and screened for a phenotype of interest. The relative abundance of each mutant in the population can be monitored over time through high-throughput sequencing of the integrated sgRNA expression cassettes. Using this technique, we outline strategies for the identification of cancer driver genes and genes mediating drug response.

Keywords: CRISPR/Cas9 mutagenesis screens; Drug sensitivity; Loss-of-function gene discovery; sgRNA libraries.

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Genome-Wide CRISPR/Cas9 Screening for Identification of Cancer Genes in Cell Lines

Affiliations

Genome-Wide CRISPR/Cas9 Screening for Identification of Cancer Genes in Cell Lines

Authors

Affiliations

Abstract

Publication types

MeSH terms

Substances

Grants and funding

LinkOut - more resources

Full Text Sources

Research Materials