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Improved vectors and genome-wide libraries for CRISPR screening

Neville E. Sanjana, Ophir Shalem, Feng Zhang
doi: https://doi.org/10.1101/006726
Neville E. Sanjana
1Broad Institute of MIT and Harvard 7 Cambridge Center Cambridge, MA 02142, USA
2McGovern Institute for Brain Research Massachusetts Institute of Technology Cambridge, MA 02139, USA
3Department of Brain and Cognitive Sciences Massachusetts Institute of Technology Cambridge, MA 02139, USA
4Department of Biological Engineering Massachusetts Institute of Technology Cambridge, MA 02139, USA
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  • For correspondence: zhang@broadinstitute.org
Ophir Shalem
1Broad Institute of MIT and Harvard 7 Cambridge Center Cambridge, MA 02142, USA
2McGovern Institute for Brain Research Massachusetts Institute of Technology Cambridge, MA 02139, USA
3Department of Brain and Cognitive Sciences Massachusetts Institute of Technology Cambridge, MA 02139, USA
4Department of Biological Engineering Massachusetts Institute of Technology Cambridge, MA 02139, USA
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  • For correspondence: zhang@broadinstitute.org
Feng Zhang
1Broad Institute of MIT and Harvard 7 Cambridge Center Cambridge, MA 02142, USA
2McGovern Institute for Brain Research Massachusetts Institute of Technology Cambridge, MA 02139, USA
3Department of Brain and Cognitive Sciences Massachusetts Institute of Technology Cambridge, MA 02139, USA
4Department of Biological Engineering Massachusetts Institute of Technology Cambridge, MA 02139, USA
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Abstract

Genome-wide, targeted loss-of-function pooled screens using the CRISPR (clustered regularly interspaced short palindrome repeats)–associated nuclease Cas9 in human and mouse cells provide an alternative screening system to RNA interference (RNAi) and have been used to reveal new mechanisms in diverse biological models1–4. Previously, we used a Genome-scale CRISPR Knock-Out (GeCKO) library to identify loss-of-function mutations conferring vemurafenib resistance in a melanoma model1. However, initial lentiviral delivery systems for CRISPR screening had low viral titer or required a cell line already expressing Cas9, limiting the range of biological systems amenable to screening.

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Posted June 28, 2014.
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Improved vectors and genome-wide libraries for CRISPR screening
Neville E. Sanjana, Ophir Shalem, Feng Zhang
bioRxiv 006726; doi: https://doi.org/10.1101/006726
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Improved vectors and genome-wide libraries for CRISPR screening
Neville E. Sanjana, Ophir Shalem, Feng Zhang
bioRxiv 006726; doi: https://doi.org/10.1101/006726

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