An optimized genome-wide, virus-free CRISPR screen for mammalian cells

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dc.contributor.authorXiong, Kaiko
dc.contributor.authorla Cour Karottki, Karen Julieko
dc.contributor.authorHefzi, Hoomanko
dc.contributor.authorLi, Songyuanko
dc.contributor.authorGrav, Lise Marieko
dc.contributor.authorLi, Shangzhongko
dc.contributor.authorSpahn, Philippko
dc.contributor.authorLee, Jae Seongko
dc.contributor.authorVentina, Ildzeko
dc.contributor.authorLee, Gyun Minko
dc.contributor.authorLewis, Nathan E.ko
dc.contributor.authorKildegaard, Helene Faustrupko
dc.contributor.authorPedersen, Lasse Ebdrupko
dc.identifier.citationCell Reports Methods, v.1, no.4-
dc.description.abstractPooled CRISPR screens have been widely applied to mammalian and other organisms to elucidate the interplay between genes and phenotypes of interest. The most popular method for delivering the CRISPR components into mammalian cells is lentivirus based. However, because lentivirus is not always an option, virus-free protocols are starting to emerge. Here, we demonstrate an improved virus-free, genome-wide CRISPR screening platform for Chinese hamster ovary cells with 75,488 gRNAs targeting 15,028 genes. Each gRNA expression cassette in the library is precisely integrated into a genomic landing pad, resulting in a very high percentage of single gRNA insertions and minimal clonal variation. Using this platform, we perform a negative selection screen on cell proliferation that identifies 1,980 genes that affect proliferation and a positive selection screen on the toxic endoplasmic reticulum stress inducer, tunicamycin, that identifies 77 gene knockouts that improve survivability.-
dc.publisherCell Press-
dc.titleAn optimized genome-wide, virus-free CRISPR screen for mammalian cells-
dc.citation.publicationnameCell Reports Methods-
dc.contributor.localauthorLee, Gyun Min-
dc.contributor.nonIdAuthorXiong, Kai-
dc.contributor.nonIdAuthorla Cour Karottki, Karen Julie-
dc.contributor.nonIdAuthorHefzi, Hooman-
dc.contributor.nonIdAuthorLi, Songyuan-
dc.contributor.nonIdAuthorGrav, Lise Marie-
dc.contributor.nonIdAuthorLi, Shangzhong-
dc.contributor.nonIdAuthorSpahn, Philipp-
dc.contributor.nonIdAuthorLee, Jae Seong-
dc.contributor.nonIdAuthorVentina, Ildze-
dc.contributor.nonIdAuthorLewis, Nathan E.-
dc.contributor.nonIdAuthorKildegaard, Helene Faustrup-
dc.contributor.nonIdAuthorPedersen, Lasse Ebdrup-
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