Construction of EZH2 Knockout Animal Model by CRISPR/Cas9 Technology / 中国肺癌杂志
Chinese Journal of Lung Cancer
;
(12): 358-364, 2018.
Article
in Chinese
| WPRIM
| ID: wpr-776309
ABSTRACT
BACKGROUND@#It has been proven that CRISPR/Cas9 (Clustered Regularly Interspaced Short Palindromic Repeats/CRISPR-associated 9) system was the modern gene-editing technology through the constitutive expression of nucleases Cas9 in the mammalian, which binds to the specific site in the genome mediated by single-guide RNA (sgRNA) at desired genomic loci. The aim of this study is that the animal model of EZH2 gene knockout was constructed using CRISPR/Cas9 technology.@*METHODS@#In this study, we designed two single-guide RNAs targeting the Exon3 and Exon4 of EZH2 gene. Then, their gene-targeting efficiency were detected by SURVEYOR assay. The lentivirus was perfused into the lungs of mice by using a bronchial tube and detected by immunohistochemistry and qRT-PCR.@*RESULTS@#The experimental results of NIH-3T3 cells verify that the designed sgEZH2 can efficiently effect the cleavage of target DNA by Cas9 in vitro. The immunohistochemistry and qRT-PCR results showed that the EZH2 expression in experimental group was significantly decreased in the mouse lung tissue.@*CONCLUSIONS@#The study successfully designed two sgRNA which can play a knock-out EZH2 function. An EZH2 knockout animal model was successfully constructed by CRISPR/Cas9 system, and it will be an effective animal model for studying the functions and mechanisms of EZH2.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Mice, Knockout
/
Gene Targeting
/
Gene Knockout Techniques
/
CRISPR-Cas Systems
/
Enhancer of Zeste Homolog 2 Protein
/
Genetics
/
Lung Neoplasms
/
Metabolism
/
Mice, Inbred C57BL
Type of study:
Qualitative research
Limits:
Animals
/
Female
/
Humans
/
Male
Language:
Chinese
Journal:
Chinese Journal of Lung Cancer
Year:
2018
Type:
Article
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