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
Dans Chinois
| 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.
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Souris knockout
/
Ciblage de gène
/
Techniques de knock-out de gènes
/
Systèmes CRISPR-Cas
/
Protéine-2 homologue de l'activateur de Zeste
/
Génétique
/
Tumeurs du poumon
/
Métabolisme
/
Souris de lignée C57BL
Type d'étude:
Recherche qualitative
Limites du sujet:
Animaux
/
Femelle
/
Humains
/
Mâle
langue:
Chinois
Texte intégral:
Chinese Journal of Lung Cancer
Année:
2018
Type:
Article
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