Molecular Inhibition of SIRT3 Pathway

3-TYP, a selective inhibitor of SIRT3, was obtained from Selleck Chemicals (Shanghai, China). Actinomycin D was purchased from APE-Bio (Houston, USA). TSA and NAM were from MedChemExpress LLC (Shanghai, China). The expressing plasmids of vIL-6-Flag and vFLIP-Flag were generated as previously described [24 (link),59 (link)]. The skeleton plasmid for lentiviral vectors vCyclin-Myc, SERBP1-HA/Myc, SIRTs-Myc and Lipt2-Flag was pCDH-CMV-MCS-EF1-copGFP (referred to as pCDH). The expressing plasmid of LANA-Flag was kindly provided by Dr. Ke Lan from Wuhan University. The small guide RNA sequences (sgRNAs) targeting SERBP1 and SIRT3 are listed in S1 Table. The control vector of all sgRNAs was Lenti-CRISPR-V2 (Lenti-V2 for short). All constructs were verified through DNA sequencing.

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Zhou J., Wang T., Zhang H., Liu J., Wei P., Xu R., Yan Q., Chen G., Li W., Gao S.J, & Lu C. (2024). KSHV vIL-6 promotes SIRT3-induced deacetylation of SERBP1 to inhibit ferroptosis and enhance cellular transformation by inducing lipoyltransferase 2 mRNA degradation. PLOS Pathogens, 20(3), e1012082.

Publication 2024

3-TYP

Corresponding Organization : UPMC Hillman Cancer Center

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Variable analysis

independent variables

3-TYP, a selective inhibitor of SIRT3
Actinomycin D
VIL-6-Flag expressing plasmid
VFLIP-Flag expressing plasmid
VCyclin-Myc expressing plasmid
SERBP1-HA/Myc expressing plasmid
SIRTs-Myc expressing plasmid
Lipt2-Flag expressing plasmid
LANA-Flag expressing plasmid
Small guide RNA (sgRNA) sequences targeting SERBP1 and SIRT3

dependent variables

Not explicitly mentioned

control variables

Lenti-CRISPR-V2 (Lenti-V2) control vector for all sgRNAs

controls

Positive control: Not specified
Negative control: Lenti-CRISPR-V2 (Lenti-V2) control vector for all sgRNAs

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