NDLI: Expansion of the CRISPR-Cas9 genome targeting space through the use of H1promoter-expressed guide

Content Provider	PubMed Central
Author	Ranganathan, Vinod Wahlin, Karl Julien, Maruotti Zack, Donald J.
Abstract	The repurposed CRISPR-Cas9 system has recently emerged as a revolutionarygenome-editing tool. Here we report a modification in the expression of the guide (gRNA)required for targeting that greatly expands the targetable genome. gRNA expression throughthe commonly used U6 promoter requires a guanosine nucleotide to initiate transcription,thus constraining genomic targeting sites to GN19NGG. We demonstrate theability to modify endogenous genes using H1 promoter-expressed gRNAs, which can be used totarget both AN19NGG and GN19NGG genomic sites. AN19NGGsites occur ~15% more frequently than GN19NGG sites in thehuman genome and the increase in targeting space is also enriched at human genes anddisease loci. Together, our results enhance the versatility of the CRISPR technology bymore than doubling the number of targetable sites within the human genome and othereukaryotic species.
Related Links	http://dx.doi.org/10.1038/ncomms5516
Ending Page	4516
Page Count	1
Starting Page	4516
File Format	PDF
ISSN	20411723
e-ISSN	20411723
Journal	Nature communications
Volume Number	5
Language	English
Publisher Date	2014-08-01
Access Restriction	Open
Subject Keyword	Biochemistry, Genetics and Molecular Biology(all) Physics and Astronomy(all) Chemistry(all) Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Chemistry Physics and Astronomy Biochemistry, Genetics and Molecular Biology

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Expansion of the CRISPR-Cas9 genome targeting space through the use of H1promoter-expressed guide–RNAs