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Enhancing the genome editing toolbox: genome wide CRISPR arrayed libraries

by Vassiliou, George S. , Skarnes, William C. , Bradley, Allan , Tzelepis, Konstantinos , Antunes, Liliana , Davis, Gregory D. , Friedrich, Mathias J , Lamberth, Jacob , Metzakopian, Emmanouil , Iyer, Vivek , Kang, Qiaohua , Davidson, Teresa , Strong, Alex , Hodgkins, Alex , Hoffmann, Christina

in 13 / 631/1647 / 631/337 / Animals / Biological research / Cell lines / Cloning / CRISPR / CRISPR-Cas Systems / Deoxyribonucleic acid / DNA / Gene Editing - methods / Gene Library / Genes / Genetic Vectors / Genome editing / Genome-Wide Association Study - methods / Genomes / GPI-Linked Proteins - metabolism / Humanities and Social Sciences / Humans / Labeling / Lentivirus - genetics / Mice / multidisciplinary / Phenotype / RNA, Guide, CRISPR-Cas Systems / Science / Science (multidisciplinary) / Signal Transduction / Transposase

2017

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Enhancing the genome editing toolbox: genome wide CRISPR arrayed libraries

2017

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Enhancing the genome editing toolbox: genome wide CRISPR arrayed libraries

2017

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Journal Article

Enhancing the genome editing toolbox: genome wide CRISPR arrayed libraries

Vassiliou, George S.,

Skarnes, William C.,

Bradley, Allan,

Tzelepis, Konstantinos,

Antunes, Liliana,

Davis, Gregory D.,

Friedrich, Mathias J,

Lamberth, Jacob,

Metzakopian, Emmanouil,

Iyer, Vivek,

Kang, Qiaohua,

Davidson, Teresa,

Strong, Alex,

Hodgkins, Alex,

Hoffmann, Christina

2017

Overview

CRISPR-Cas9 technology has accelerated biological research becoming routine for many laboratories. It is rapidly replacing conventional gene editing techniques and has high utility for both genome-wide and gene-focussed applications. Here we present the first individually cloned CRISPR-Cas9 genome wide arrayed sgRNA libraries covering 17,166 human and 20,430 mouse genes at a complexity of 34,332 sgRNAs for human and 40,860 sgRNAs for the mouse genome. For flexibility in generating stable cell lines the sgRNAs have been cloned in a lentivirus backbone containing PiggyBac transposase recognition elements together with fluorescent and drug selection markers. Over 95% of tested sgRNA induced specific DNA cleavage as measured by CEL-1 assays. Furthermore, sgRNA targeting GPI anchor protein pathway genes induced loss of function mutations in human and mouse cell lines measured by FLAER labelling. These arrayed libraries offer the prospect for performing screens on individual genes, combinations as well as larger gene sets. They also facilitate rapid deconvolution of signals from genome-wide screens. This set of vectors provide an organized comprehensive gene editing toolbox of considerable scientific value.

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Publisher

Nature Publishing Group UK,Nature Publishing Group,Nature Portfolio

Subject

/ 631/1647

/ 631/337

/ Animals

/ Biological research

/ Cell lines

/ Cloning

/ CRISPR