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In vivo genome editing using Staphylococcus aureus Cas9
#MMPMID25830891
Ran FA
; Cong L
; Yan WX
; Scott DA
; Gootenberg JS
; Kriz AJ
; Zetsche B
; Shalem O
; Wu X
; Makarova KS
; Koonin EV
; Sharp PA
; Zhang F
Nature
2015[Apr]; 520
(7546
): 186-91
PMID25830891
show ga
The RNA-guided endonuclease Cas9 has emerged as a versatile genome-editing
platform. However, the size of the commonly used Cas9 from Streptococcus pyogenes
(SpCas9) limits its utility for basic research and therapeutic applications that
use the highly versatile adeno-associated virus (AAV) delivery vehicle. Here, we
characterize six smaller Cas9 orthologues and show that Cas9 from Staphylococcus
aureus (SaCas9) can edit the genome with efficiencies similar to those of SpCas9,
while being more than 1 kilobase shorter. We packaged SaCas9 and its single guide
RNA expression cassette into a single AAV vector and targeted the cholesterol
regulatory gene Pcsk9 in the mouse liver. Within one week of injection, we
observed >40% gene modification, accompanied by significant reductions in serum
Pcsk9 and total cholesterol levels. We further assess the genome-wide targeting
specificity of SaCas9 and SpCas9 using BLESS, and demonstrate that
SaCas9-mediated in vivo genome editing has the potential to be efficient and
specific.
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