Lentivirus based control and gene specific small hairpin rnas shrnas

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Lentivirus-based control and gene-specific small hairpin RNAs (shRNAs) are laboratory tools used for gene silencing and knockdown experiments. These products provide a reliable and efficient method for the delivery and expression of short hairpin RNA sequences in cells, enabling the targeted suppression of specific gene expression.

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Lentivirus-based small hairpin RNAs (shRNA) Lentivirus-based control and gene-specific shRNAs Small hairpin RNA (shRNA) Lentivirus-based shRNAs ShRNA lentiviruses Specific shRNAs Control shRNA Control lentivirus RNases Lentivirus

3 protocols using lentivirus based control and gene specific small hairpin rnas shrnas

Lentiviral Transduction of Prostate Cancer Cells

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Lentivirus-based control and gene-specific small hairpin RNAs (shRNAs) were purchased from Sigma-Aldrich. Viral packaging plasmids (pEXQV and pVSV-G) and shRNA plasmid were transfected to 293T cells by using Lipofectamine 2000. After 24 h, virus culture medium was replaced with DMEM containing 10% FBS with 1:100 of sodium Pyruvate. 48 h post transfection, medium was collected and added to prostate cancer cells added with 12 μg/ml of polybrene. Prostate cancer cells were harvested 48 h after puromycin selection. shRNA sequence information is provided in Table S2.

Jin X., Ding D., Yan Y., Li H., Wang B., Ma L., Ye Z., Ma T., Wu Q., Rodrigues D.N., Kohli M., Jimenez R., Wang L., Goodrich D.W., de Bono J., Dong H., Wu H., Zhu R, & Huang H. (2018). Phosphorylated RB Promotes Cancer Immunity by Inhibiting NF-κB Activation and PD-L1 Expression. Molecular cell, 73(1), 22-35.e6.

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Lentiviral shRNA Transduction in Prostate Cancer

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Jin X., Yan Y., Wang D., Ding D., Ma T., Ye Z., Jimenez R., Wang L., Wu H, & Huang H. (2018). DUB3 promotes BET inhibitor resistance and cancer progression through deubiquitinating BRD4. Molecular cell, 71(4), 592-605.e4.

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Lentiviral Transduction for shRNA and CRISPR Studies

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Lentivirus-based control and gene-specific small hairpin RNAs (shRNAs) were purchased from Sigma-Aldrich. Lipofectamine 2000 was used to transfect 293T cells with shRNA plasmids and viral packaging plasmids (pVSV-G and pEXQV). At 24 hours after transfection, the medium was replaced with fresh DMEM, containing 10% FBS and 1 mM of sodium pyruvate. 48 hours after transfection, the virus culture medium was collected, filtered and added to cancer cells supplemented with 12 µg/mL of polybrene. At 24 hours after infection, the infected cells were selected with 10 µg /mL of puromycin. The shRNA and sgRNA sequence is provided in the Supplementary Table 2.

Wang B., Zhou Y., Zhang J., Jin X., Wu H, & Huang H. (2020). Fructose-1,6-bisphosphatase loss modulates STAT3-dependent expression of PD-L1 and cancer immunity. Theranostics, 10(3), 1033-1045.

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