Sirnas

Manufactured by Santa Cruz Biotechnology

Sourced in United States

SiRNAs (small interfering RNAs) are a class of double-stranded RNA molecules that play a key role in the RNA interference (RNAi) pathway. They function by targeting and silencing specific messenger RNA (mRNA) molecules, thereby preventing the expression of the corresponding genes.

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Lab products found in correlation

95 protocols using sirnas

Breast Cancer Cell siRNA Transfection

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SiRNA transfection was performed as previously described [39 (link)]. Briefly, the indicated breast cancer cells were randomly seeded in 60 mm dishes for 24 h. RPMI opti-MEM, lipofectamine RNAiMax (Invitrogen) reagent and siRNAs(Santa Cruz, CA) targeting human USP14/UCHL5/AR or siRNAs(Santa Cruz, CA) with non-specific sequences mixtures were prepared respectively, then the mixtures was added in each group and cultured for 72 h. Fresh medium was replaced appropriately after transfection for 6 h.

Liao Y., Xia X., Liu N., Cai J., Guo Z., Li Y., Jiang L., Dou Q.P., Tang D., Huang H, & Liu J. (2018). Growth arrest and apoptosis induction in androgen receptor-positive human breast cancer cells by inhibition of USP14-mediated androgen receptor deubiquitination. Oncogene, 37(14), 1896-1910.

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Engineered KITENIN constructs for study

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Expression constructs were generated by PCR‐based methods: V5‐tagged, Myc‐tagged, HA‐tagged KITENIN, GST‐tagged deletion mutants of KITENIN, Δ463–471‐KITENIN, and His‐tagged KITENIN. All constructs were confirmed by sequencing. pEGFP‐N1‐RACK1 was a gift from Anna Huttenlocher (Addgene plasmid #41088). All siRNAs used for gene silencing were obtained from Santa Cruz Biotechnology. Each consisted of a mixture of several sequences, thus eliminating sequence‐specific diversity.

Kim S.J., Sun E.G., Bae J.A., Park S., Hong C., Park Z., Kim H, & Kim K.K. (2022). A peptide interfering with the dimerization of oncogenic KITENIN protein and its stability suppresses colorectal tumour progression. Clinical and Translational Medicine, 12(7), e871.

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Silencing of MFN2 and Parkin in BMDMs

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Silencing of MFN2 and Parkin was achieved using small interfering RNAs (siRNAs) (200 nM) for mouse MFN2 mRNA target sequences (Santa Cruz Biotechnology), mouse Parkin mRNA target sequences (Santa Cruz Biotechnology), and negative control siRNAs (Bioneer, Daejeon, Korea). The siRNA oligonucleotides were transfected into cultured BMDMs using Lipofectamine 3000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions.

Lee J., Choi J.A., Cho S.N., Son S.H, & Song C.H. (2019). Mitofusin 2-Deficiency Suppresses Mycobacterium tuberculosis Survival in Macrophages. Cells, 8(11), 1355.

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Bcl-2/Bcl-xL Knockdown via siRNA Transfection

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Transfection of siRNA was performed using Lipofectamine^TM RNAiMAX (Invitrogen, Grand Island, NY, USA) according to the manufacturer's instructions. siRNAs targeting Bcl-2 and Bcl-xL were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA) and Invitrogen, respectively. Control siRNA (#6568) was purchased from Cell Signaling Technology (Danvers, MA, USA). The transfected cells were used for the experiments 3 days after siRNA transfection.

Tamaki H., Harashima N., Hiraki M., Arichi N., Nishimura N., Shiina H., Naora K, & Harada M. (2014). Bcl-2 family inhibition sensitizes human prostate cancer cells to docetaxel and promotes unexpected apoptosis under caspase-9 inhibition. Oncotarget, 5(22), 11399-11412.

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Stable Transfection of KYNU Gene

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The transfection of pcDNA‐KYNU (OriGene #RC214932) was performed by using the TransIT‐LT1 transfection reagent (Mirus) according to manufacturer's recommendations. Stable transfection was achieved by G418 selection. For siRNA experiment, siRNAs was purchased from Santa Cruz (#sc‐51370).

Liu Y., Feng X., Lai J., Yi W., Yang J., Du T., Long X., Zhang Y, & Xiao Y. (2019). A novel role of kynureninase in the growth control of breast cancer cells and its relationships with breast cancer. Journal of Cellular and Molecular Medicine, 23(10), 6700-6707.

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Targeted Gene Silencing in A549 Cells

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A549 cells were plated in 60-mm dishes at ∼70-80% confluence and transfected with siRNAs (Santa Cruz Biotechnology, Santa Cruz, CA) using Lipofectamine 2000 (Invitrogen, Grand Island, NY) as described [40 (link)]. The siRNAs are pools of at least three different siRNA oligoes that are specific to the target. Negative control siRNA (siNeg, Santa Cruz Biotechnology) was used as a negative control. Two days after transfection, cells were lysed for Western blot analysis.

Zhou Q., Dai J., Chen T., Dada L.A., Zhang X., Zhang W., DeCamp M.M., Winn R.A., Sznajder J.I, & Zhou G. (2017). Downregulation of PKCζ/Pard3/Pard6b is responsible for lung adenocarcinoma cell EMT and invasion. Cellular signalling, 38, 49-59.

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Molecular Tools for Studying Cell Signaling

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Metafectene Pro transfection reagent was obtained from Biontex. siRNAs targeting 14-3-3σ (sc-29590), YAP1 (sc-38637) were purchased from Santa Cruz Biotechnology. Antibodies against GFP (ab290), YAP1 (ab52771), pYAP1 (ab76252) were from Abcam. Antibodies against 14-3-3σ (05-632), RRM1 (MABE567) and ChIP Assay kit (17-295) were purchased from EMD Millipore. Antibody against RRM2 was generated in-house [31 (link)]. Lipofectamine, pcDNA3.1(+) plasmid, and G418 were from Invitrogen. RNeasy Mini kit and Qiagen Blood and Cell Culture DNA Kit were from Qiagen. The iScript™ cDNA synthesis kit and the SYBR Green PCR master mix were from Bio-Rad and Applied Biosystems, respectively. Gemcitabine were purchased from Besse Medical

Qin L., Dong Z, & Zhang J.T. (2016). 14-3-3σ regulation of and interaction with YAP1 in acquired gemcitabine resistance via promoting ribonucleotide reductase expression. Oncotarget, 7(14), 17726-17736.

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Silencing PHD Isoforms in Cells

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siRNAs that targeted PHD1, PHD2, PHD3 and a control siRNA were purchased from Santa Cruz Biotechnology. Cells were transfected using Lipofectamine RNAiMAX (Invitrogen) according to manufacturer's instructions. The efficiency of siRNA was determined by semi-quantitative nested RT-PCR of the relevant genes using gene-specific primer pairs purchased from Santa Cruz Biotechnology.

Hou P., Kuo C.Y., Cheng C.T., Liou J.P., Ann D.K, & Chen Q. (2014). Intermediary Metabolite Precursor Dimethyl-2-Ketoglutarate Stabilizes Hypoxia-Inducible Factor-1α by Inhibiting Prolyl-4-Hydroxylase PHD2. PLoS ONE, 9(11), e113865.

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Efficient siRNA Transfection in Cells

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All small interfering RNAs (siRNAs) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). The nonsilencing control siRNA was used as a negative control. Scrambled siRNA-treated cells were also used as controls. The transfection of siRNA was performed in Opti-MEM I Reduced-Serum Medium using Lipofectamine RNAiMAX (Invitrogen) [24 (link)]. We confirmed the expression of a target gene by Western-blot. There are differences in the suppression ratio (namely reflecting the transfection efficiency) according to target genes. In siRNA of eNOS, the suppression ratio was approximately 99%. In siRNA of p22phox, it was 96.5%. There is a report that the transfection reagent (RNAiMAX), which we used in the present study, shows the high transfection efficiency in smooth muscle cell as adherent cell, too [25 (link)].

Maeda M., Hayashi T., Mizuno N., Hattori Y, & Kuzuya M. (2015). Intermittent High Glucose Implements Stress-Induced Senescence in Human Vascular Endothelial Cells: Role of Superoxide Production by NADPH Oxidase. PLoS ONE, 10(4), e0123169.

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siRNA-mediated Knockdown of rogdi

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siRNAs (Santa Cruz, sc-93538) were used to silence rogdi according to the protocol provided by the manufacturer. siRNA (7.5 μl) and 250-μl Opti-MEM (Gibco, 11058021) were mixed. Lipofectamine 2000 (Invitrogen, 11668-019) (7.5 μl) was added to the other Opti-MEM (250 μl) mixture and mixed for 5 min. The diluted siRNA and Lipofectamine were mixed for 20 min. The reagents were added into 6-well plates in which cells had been seeded (5 × 10⁵ cells/well) for 16 h. Control cells were treated with Stealth RNAi Negative Control Duplex (Invitrogen, 12935300).

Chen Y.F., Cho J.J., Huang T.H., Tseng C.N., Huang E.Y, & Cho C.L. (2016). Downregulation of a novel human gene, ROGDI, increases radiosensitivity in cervical cancer cells. Cancer Biology & Therapy, 17(10), 1070-1078.

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