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FEBS J. 2016 Sep;283(17):3232-8. doi: 10.1111/febs.13777. Epub 2016 Jun 22.

CRISPR guide RNA design for research applications.

Author information

1
Drosophila RNAi Screening Center, Harvard Medical School, Boston, MA, USA.
2
Department of Genetics, Harvard Medical School, Boston, MA, USA.
3
Howard Hughes Medical Institute, Boston, MA, USA.

Abstract

The rapid rise of CRISPR as a technology for genome engineering and related research applications has created a need for algorithms and associated online tools that facilitate design of on-target and effective guide RNAs (gRNAs). Here, we review the state of the art in CRISPR gRNA design for research applications of the CRISPR-Cas9 system, including knockout, activation, and inhibition. Notably, achieving good gRNA design is not solely dependent on innovations in CRISPR technology. Good design and design tools also rely on availability of high-quality genome sequence and gene annotations, as well as on availability of accumulated data regarding off-targets and effectiveness metrics.

KEYWORDS:

CRISPR-Cas9; CRISPRa; CRISPRi; bioinformatics; gene editing; genome annotation; genome engineering

PMID:
27276584
PMCID:
PMC5014588
DOI:
10.1111/febs.13777
[Indexed for MEDLINE]
Free PMC Article

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