CRISPR-Cas9 Mediated Hsp90 Knockout

Hsp90AA1 sgRNA sequences and the siHsp90α and siHsp90β sequences were synthesized by iGeneBio (Guangzhou, China). The transfection of CRISPR-Cas9 plasmids for Hsp90α knockout, siHsp90α and siHsp90β were carried out with LIPO3000 (L3000015, Life Technologies, USA). According to the protocol, after 48 h, cells were collected, and incubated in the selection medium containing 800 μg/mL G418 (G001, MDBio, Inc, China) for 2–3 weeks. The successfully Hsp90AA1 knockout clones were confirmed by western blotting (21 (link)).

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Liu L., Deng Y., Zheng Z., Deng Z., Zhang J., Li J., Liang M., Zhou X., Tan W., Yang H., Neckers L.M., Zou F, & Chen X. (2021). Hsp90 Inhibitor STA9090 Sensitizes Hepatocellular Carcinoma to Hyperthermia-Induced DNA Damage by Suppressing DNA-PKcs Protein Stability and mRNA Transcription. Molecular cancer therapeutics, 20(10), 1880-1892.

Publication 2021

LIPO3000

Cells Clones Crispr G418 Hsp90aa1 Plasmids Transfection

Corresponding Organization : Southern Medical University

Other organizations : Nanfang Hospital, Center for Cancer Research

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Variable analysis

independent variables

Transfection of CRISPR-Cas9 plasmids for Hsp90α knockout
SiHsp90α
SiHsp90β

dependent variables

Hsp90α knockout clones confirmed by western blotting

control variables

LIPO3000 (L3000015, Life Technologies, USA) for transfection
Selection medium containing 800 μg/mL G418 (G001, MDBio, Inc, China) for 2-3 weeks

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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