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A new age in functional genomics using CRISPR/Cas9 in arrayed library screening

Agrotis, A; Ketteler, R; (2015) A new age in functional genomics using CRISPR/Cas9 in arrayed library screening. Front Genet , 6 , Article 00300. 10.3389/fgene.2015.00300. Green open access

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Abstract

CRISPR technology has rapidly changed the face of biological research, such that precise genome editing has now become routine for many labs within several years of its initial development. What makes CRISPR/Cas9 so revolutionary is the ability to target a protein (Cas9) to an exact genomic locus, through designing a specific short complementary nucleotide sequence, that together with a common scaffold sequence, constitute the guide RNA bridging the protein and the DNA. Wild-type Cas9 cleaves both DNA strands at its target sequence, but this protein can also be modified to exert many other functions. For instance, by attaching an activation domain to catalytically inactive Cas9 and targeting a promoter region, it is possible to stimulate the expression of a specific endogenous gene. In principle, any genomic region can be targeted, and recent efforts have successfully generated pooled guide RNA libraries for coding and regulatory regions of human, mouse and Drosophila genomes with high coverage, thus facilitating functional phenotypic screening. In this review, we will highlight recent developments in the area of CRISPR-based functional genomics and discuss potential future directions, with a special focus on mammalian cell systems and arrayed library screening.

Type: Article
Title: A new age in functional genomics using CRISPR/Cas9 in arrayed library screening
Location: Switzerland
Open access status: An open access version is available from UCL Discovery
DOI: 10.3389/fgene.2015.00300
Publisher version: http://dx.doi.org/10.3389/fgene.2015.00300
Additional information: Copyright © 2015 Agrotis and Ketteler. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
Keywords: CRISPR, Cas9/sgRNA, high-content imaging, high-throughput screening, knockdown, siRNA
UCL classification: UCL
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences
UCL > Provost and Vice Provost Offices > School of Life and Medical Sciences > Faculty of Life Sciences > Lab for Molecular Cell Bio MRC-UCL
URI: https://discovery.ucl.ac.uk/id/eprint/1473508
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