Genome-wide CRISPR screen in a mouse model of tumor growth and metastasis

By Sidi Chen, Neville E. Sanjana, K. Zheng, Ophir Shalem, Kyungheon Lee, Xi Shi, David A. Scott, Jun S. Song, Jen Q. Pan, Ralph Weissleder and 3 more. Cell, 2015

846 citations recorded by the source corpus.

Summary Genetic screens are powerful tools for identifying genes responsible for diverse phenotypes. Here we describe a genome-wide CRISPR-Cas9-mediated loss-of-function screen in tumor growth and metastasis. We mutagenized a non-metastatic mouse cancer cell line using a genome-scale library with 67,405 single guide RNAs (sgRNAs). The mutant cell pool rapidly generates metastases when transplanted into immunocompromised mice. Enriched sgRNAs in lung metastases and late stage primary tumors were found to target a small set of genes, suggesting specific loss-of-function mutations drive tumor growth and metastasis. Individual sgRNAs and a small pool of 624 sgRNAs targeting the top scoring genes from the primary screen dramatically accelerate metastasis. In all of these experiments, the effect of mutations on primary tumor growth positively correlates with the development of metastases. Our study demonstrates Cas9-based screening as a robust method to systematically assay gene phenotypes in cancer evolution in vivo.

Subfields: Molecular Biology

View Paper: https://www.semanticscholar.org/paper/953e4884465d48324d550c5559a57da2928c2239

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Citations in the corpus

Compact and highly active next-generation libraries for CRISPR-mediated gene repression and activation (2016, methodology): “, 2015), or for in vivo screens where cell engraftment and library representation may represent a limiting factor (Braun et al., 2016; Chen et al., 2015).”
Quality control, modeling, and visualization of CRISPR screens with MAGeCK-VISPR (2015, background): “Conclusion The recently developed CRISPR screening is a powerful technology in functional studies with different foci, including tumor progression and metastasis [29], drug resistance [3], immune response [30], and stem cell differentiation [4].”
Applications of genome editing technology in the targeted therapy of human diseases: mechanisms, advances and prospects (2020, methodology): “Another successful example involved the use of CRISPR/Cas9-mediated loss-of-function screening to identify cancer metastasis-related genes.(240) In this study, a nonmetastatic lung cancer cell line was infected with the mouse genome-scale CRISPR knockout (mGeCKO) sgRNA library and subcutaneously tra”
Dissecting Immune Circuits by Linking CRISPR-Pooled Screens with Single-Cell RNA-Seq. (2016, neutral): “These screens are usually used to identify crude cellular phenotypes, such as survival, cell shape, or single gene markers (Chen et al., 2015; Hsu et al., 2014; Shalem et al., 2015).”
A major chromatin regulator determines resistance of tumor cells to T cell–mediated killing (2018, methodology): “We calculated the percentage of cell survival for each viral concentration using the following formula (39):”
High-Resolution CRISPR Screens Reveal Fitness Genes and Genotype-Specific Cancer Liabilities. (2015, positive): “…whose loss of function enables drug and toxin resistance (Koike-Yusa et al., 2014; Shalem et al., 2014; Wang et al., 2014), accelerates metastasis (Chen et al., 2015), or influences immune response (Parnas et al., 2015) confirm the vast improvement that the CRISPR technology offers in mammalian…”
Systems Biology of Cancer Metastasis (2019, background): “Developments in gene editing have facilitated large-scale in vivo studies of the genetic alterations that drive cancer tumor growth and metastasis(Chen et al., 2015; Kalhor et al., 2018).”
Synergistic drug combinations for cancer identified in a CRISPR screen for pairwise genetic interactions (2017, positive): “In previous studies, we 15 and others 46,47 have seen that measurement of smaller libraries at higher coverage reduced false positives/false negatives from larger primary screens.”