CRISPR guide RNA design for research applications.
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.
Drosophila RNAi Screening Center, Harvard Medical School, Boston, MA, USA.|Department of Genetics, Harvard Medical School, Boston, MA, USA.
Stephanie E
SE
Mohr
Drosophila RNAi Screening Center, Harvard Medical School, Boston, MA, USA.|Department of Genetics, Harvard Medical School, Boston, MA, USA.
Yanhui
Y
Hu
Department of Genetics, Harvard Medical School, Boston, MA, USA.
Benjamin
B
Ewen-Campen
Department of Genetics, Harvard Medical School, Boston, MA, USA.
Benjamin E
BE
Housden
Department of Genetics, Harvard Medical School, Boston, MA, USA.
Raghuvir
R
Viswanatha
Drosophila RNAi Screening Center, Harvard Medical School, Boston, MA, USA.|Department of Genetics, Harvard Medical School, Boston, MA, USA.|Howard Hughes Medical Institute, Boston, MA, USA.
Norbert
N
Perrimon
D000465:Algorithms; D000818:Animals; D064113:CRISPR-Cas Systems; D055786:Gene Knockout Techniques; D018390:Gene Targeting; D005818:Genetic Engineering / Q000379:methods; D006801:Humans; D058977:Molecular Sequence Annotation; D000094704:RNA, Guide, CRISPR-Cas Systems / Q000235:genetics*
D000094704:RNA, Guide, CRISPR-Cas Systems
CRISPR-Cas9; CRISPRa; CRISPRi; bioinformatics; gene editing; genome annotation; genome engineering