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.
Artigo
em Chinês
| 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.
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Camundongos Knockout
/
Marcação de Genes
/
Técnicas de Inativação de Genes
/
Sistemas CRISPR-Cas
/
Proteína Potenciadora do Homólogo 2 de Zeste
/
Genética
/
Neoplasias Pulmonares
/
Metabolismo
/
Camundongos Endogâmicos C57BL
Tipo de estudo:
Pesquisa qualitativa
Limite:
Animais
/
Feminino
/
Humanos
/
Masculino
Idioma:
Chinês
Revista:
Chinese Journal of Lung Cancer
Ano de publicação:
2018
Tipo de documento:
Artigo
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