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2018 ; 2018
(ä): 9576959
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User-Friendly Genetic Conditional Knockout Strategies by CRISPR/Cas9
#MMPMID30013601
Chen L
; Ye Y
; Dai H
; Zhang H
; Zhang X
; Wu Q
; Zhu Z
; Spalinskas R
; Ren W
; Zhang W
Stem Cells Int
2018[]; 2018
(ä): 9576959
PMID30013601
show ga
Loss-of-function studies are critically important in gene functional analysis of
model organisms and cells. However, conditional gene inactivation in diploid
cells is difficult to achieve, as it involves laborious vector construction,
multifold electroporation, and complicated genotyping. Here, a strategy is
presented for generating biallelic conditional gene and DNA regulatory region
knockouts in mouse embryonic stem cells by codelivery of CRISPR-Cas9 and
short-homology-arm targeting vectors sequentially or simultaneously.
Collectively, a simple and rapid method was presented to knock out any DNA
element conditionally. This approach will facilitate the functional studies of
essential genes and regulatory regions during development.