Construction and characterization of QKI knockout GC1-spg cell strain with CRISPR/CAS9 / 基础医学与临床
Basic & Clinical Medicine
;
(12): 589-593, 2018.
Article
in Chinese
| WPRIM
| ID: wpr-693947
ABSTRACT
Objective To investigate whether QKI protein plays any important role in the process of spermatogene-sis.Constructing GC1-spg cell strain which knocked out QKI by the technology of CRISPR/Cas9,and detecting its effect on the proliferation and differentiation of QKI protein in vitro.Methods The plasmid PX330 was used to construct QKI knockout recombinant plasmid, then transfected it to GC1-spg wild-type cells and selected by puromycin.GC1-spg knock-QKI cell strain was identified by Western blot and gene sequencing; The wild-type and knockout cell strain was cultured normally,then detected the growth curve by cell counting kit(CCK8),and using quantitative PCR to get the changes of meiotic-related gene differentiation.Results QKI knockout GC1-spg cell strain was successfully constructed.Compared with the control group,the growth of QKI knockout cell strain was significantly decreased(P<0.05), and the expression of meiosis related molecular marker gene of c-kit, Mtl5 and Pspa2 was significantly decreased(P<0.05).Conclusions QKI proteins can affect reproductive sper-matogenesis by acting on proliferation and differentiation.
Full text:
Available
Index:
WPRIM (Western Pacific)
Language:
Chinese
Journal:
Basic & Clinical Medicine
Year:
2018
Type:
Article
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