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Scientific Fields
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Published in BMC genomics - 06 Feb 2019

Boettcher M, Covarrubias S, Biton A, Blau J, Wang H, Zaitlen N, McManus MT

Link to Pubmed [PMID] – 30727954

BMC Genomics 2019 Feb;20(1):107

BACKGROUND: While pooled loss- and gain-of-function CRISPR screening approaches have become increasingly popular to systematically investigate mammalian gene function, the large majority of them have thus far not investigated the influence of cellular heterogeneity on screen results. Instead most screens are analyzed by averaging the abundance of perturbed cells from a bulk population of cells.

RESULTS: Here we developed multi-level barcoded sgRNA libraries to trace multiple clonal Cas9 cell lines exposed to the same environment. The first level of barcoding allows monitoring growth kinetics and treatment responses of multiplexed clonal cell lines under identical conditions while the second level enables in-sample replication and tracing of sub-clonal lineages of cells expressing the same sgRNA.

CONCLUSION: Using our approach, we illustrate how heterogeneity in growth kinetics and treatment response of clonal cell lines impairs the results of pooled genetic screens.

https://www.ncbi.nlm.nih.gov/pubmed/30727954