Expansion of the CRISPR-Cas9 genome targeting space through the use of H1 promoter-expressed guide–RNAs View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2014-08-08

AUTHORS

Vinod Ranganathan, Karl Wahlin, Julien Maruotti, Donald J. Zack

ABSTRACT

The repurposed CRISPR-Cas9 system has recently emerged as a revolutionary genome-editing tool. Here we report a modification in the expression of the guide RNA (gRNA) required for targeting that greatly expands the targetable genome. gRNA expression through the commonly used U6 promoter requires a guanosine nucleotide to initiate transcription, thus constraining genomic-targeting sites to GN19NGG. We demonstrate the ability to modify endogenous genes using H1 promoter-expressed gRNAs, which can be used to target both AN19NGG and GN19NGG genomic sites. AN19NGG sites occur ~15% more frequently than GN19NGG sites in the human genome and the increase in targeting space is also enriched at human genes and disease loci. Together, our results enhance the versatility of the CRISPR technology by more than doubling the number of targetable sites within the human genome and other eukaryotic species. More... »

PAGES

4516-4516

References to SciGraph publications

  • 2010-09-07. Non-muscle myosin II regulates survival threshold of pluripotent stem cells in NATURE COMMUNICATIONS
  • 2013-08-01. CAS9 transcriptional activators for target specificity screening and paired nickases for cooperative genome engineering in NATURE BIOTECHNOLOGY
  • 2009-04-12. Enzymatic assembly of DNA molecules up to several hundred kilobases in NATURE METHODS
  • 2013-08-11. High-throughput profiling of off-target DNA cleavage reveals RNA-programmed Cas9 nuclease specificity in NATURE BIOTECHNOLOGY
  • 2010-06-25. A Rapid and General Assay for Monitoring Endogenous Gene Modification in ENGINEERED ZINC FINGER PROTEINS
  • 2013-10-17. CRISPR interference (CRISPRi) for sequence-specific control of gene expression in NATURE PROTOCOLS
  • 2013-07-21. DNA targeting specificity of RNA-guided Cas9 nucleases in NATURE BIOTECHNOLOGY
  • 2013-06-23. High frequency off-target mutagenesis induced by CRISPR-Cas nucleases in human cells in NATURE BIOTECHNOLOGY
  • 2012-04-08. FLASH Assembly of TALENs Enables High-Throughput Genome Editing in NATURE BIOTECHNOLOGY
  • 2002-05. Expanding small RNA interference in NATURE BIOTECHNOLOGY
  • 2009-08-13. Efficient targeting of expressed and silent genes in human ESCs and iPSCs using zinc-finger nucleases in NATURE BIOTECHNOLOGY
  • 2012-01-05. A transcription activator-like effector toolbox for genome engineering in NATURE PROTOCOLS
  • 2010-12-12. Selection-Free Zinc-Finger Nuclease Engineering by Context-Dependent Assembly (CoDA) in NATURE METHODS
  • 2011-01-19. Programmable Sequence-Specific Transcriptional Regulation of Mammalian Genome Using Designer TAL Effectors in NATURE BIOTECHNOLOGY
  • 2010-12-22. A TALE nuclease architecture for efficient genome editing in NATURE BIOTECHNOLOGY
  • 2013-01-29. Efficient In Vivo Genome Editing Using RNA-Guided Nucleases in NATURE BIOTECHNOLOGY
  • 2007-07-01. An improved zinc-finger nuclease architecture for highly specific genome editing in NATURE BIOTECHNOLOGY
  • 2013-01-29. Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease in NATURE BIOTECHNOLOGY
  • 2013-08-08. Targeted genome modification of crop plants using a CRISPR-Cas system in NATURE BIOTECHNOLOGY
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    DOI

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    DIMENSIONS

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    49 schema:description The repurposed CRISPR-Cas9 system has recently emerged as a revolutionary genome-editing tool. Here we report a modification in the expression of the guide RNA (gRNA) required for targeting that greatly expands the targetable genome. gRNA expression through the commonly used U6 promoter requires a guanosine nucleotide to initiate transcription, thus constraining genomic-targeting sites to GN19NGG. We demonstrate the ability to modify endogenous genes using H1 promoter-expressed gRNAs, which can be used to target both AN19NGG and GN19NGG genomic sites. AN19NGG sites occur ~15% more frequently than GN19NGG sites in the human genome and the increase in targeting space is also enriched at human genes and disease loci. Together, our results enhance the versatility of the CRISPR technology by more than doubling the number of targetable sites within the human genome and other eukaryotic species.
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