Sendai virus, an RNA virus with no risk of genomic integration, delivers CRISPR/Cas9 for efficient gene editing.
The advent of RNA-guided endonuclease (RGEN)-mediated gene editing, specifically via CRISPR/Cas9, has spurred intensive efforts to improve the efficiency of both RGEN delivery and targeted mutagenesis. The major viral vectors in use for delivery of Cas9 and its associated guide RNA, lentiviral and adeno-associated viral systems, have the potential for undesired random integration into the host genome. Here, we repurpose Sendai virus, an RNA virus with no viral DNA phase and that replicates solely in the cytoplasm, as a delivery system for efficient Cas9-mediated gene editing. The high efficiency of Sendai virus infection resulted in high rates of on-target mutagenesis in cell lines (75-98% at various endogenous and transgenic loci) and primary human monocytes (88% at the ccr5 locus) in the absence of any selection. In conjunction with extensive former work on Sendai virus as a promising gene therapy vector that can infect a wide range of cell types including hematopoietic stem cells, this proof-of-concept study opens the door to using Sendai virus as well as other related paramyxoviruses as versatile and efficient tools for gene editing.
Department of Microbiology, Icahn School of Medicine at Mount Sinai , New York, New York, USA.
Arnold
A
Park
Department of Microbiology, Icahn School of Medicine at Mount Sinai , New York, New York, USA.
Patrick
P
Hong
Department of Microbiology, Icahn School of Medicine at Mount Sinai , New York, New York, USA.
Sohui T
ST
Won
Department of Microbiology, Icahn School of Medicine at Mount Sinai , New York, New York, USA.
Patricia A
PA
Thibault
Department of Microbiology, Icahn School of Medicine at Mount Sinai , New York, New York, USA.
Frederic
F
Vigant
Department of Microbiology, Icahn School of Medicine at Mount Sinai , New York, New York, USA.
Kasopefoluwa Y
KY
Oguntuyo
Department of Microbiology, Icahn School of Medicine at Mount Sinai , New York, New York, USA.
Justin D
JD
Taft
Department of Microbiology, Icahn School of Medicine at Mount Sinai , New York, New York, USA.
Benhur
B
Lee