稳定敲降NF-YB基因HeLa细胞系的构建

doi:10.3969/j.issn.1004-616x.2020.06.001

Abstract

Abstract: OBJECTIVE: To construct a stably knockdown NF-YB gene in HeLa cells using the Lentivirus-mediated RNA interference (RNAi) technique. METHODS: shRNA-NF-YB primers were designed and the H1/GFP-Puro-shRNA-NF-YB plasmids were constructed by molecular cloning. The virus-infected HeLa cells were collected after transfection with 293T. Stably knock down cells were obtained by screening cells with puromycin. Cell lines were identified by Western blot and qPCR. RESULTS: The constructed plasmid was sequenced by the company and was consistent with the designed primer sequence. Western blot and qPCR results show that inhibition of NF-YB protein and mRNA expression was significant. CONCLUSION: In this experiment,NF-YB-shRNA-HeLa cell line was successfully constructed and HeLa cells with specific NF-YB gene silencing knockdown were obtained.

Key words: NF-YB gene, RNA interference, HeLa cells, lentivirus-mediated

CLC Number:

Q813

DU Lu, JIANG Xiaoyan, DING Kuke. Stable knockdown of NF-YB gene in the HeLa cell line[J]. Carcinogenesis, Teratogenesis & Mutagenesis, 2020, 32(6): 409-413.

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Stable knockdown of NF-YB gene in the HeLa cell line

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