Control shrna

Manufactured by Santa Cruz Biotechnology

Sourced in United States

Control shRNA is a laboratory reagent used in gene silencing experiments. It functions as a non-targeting control, allowing researchers to assess the effects of the experimental shRNA on gene expression without the influence of target-specific knockdown.

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Control shRNA lentiviral particles (38 citations) Control shRNA lentiviral particles-A (10 citations) Control shRNA lentiviral particles (sc-108080) (9 citations) Control shRNA lentiviral particles-A (sc-108080) (7 citations) Control shRNA lentivirus (5 citations) Control shRNA (sc-108080) (5 citations) Control shRNA (h) lentiviral particles-A (2 citations) Scrambled shRNA control (sc-108080) lentiviral particles (2 citations) Control (sc‐108080) short hairpin RNA (shRNA) lentiviral particles (2 citations) Sc-108060 control shRNA plasmid-A (2 citations)

41 protocols using control shrna

Investigating miR-663b and CD99 in Jurkat Cells

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Jurkat cells were transfected with inhibitor control (Guangzhou Ribobio Co., Ltd., Guangzhou, China), miR-663b inhibitor (Guangzhou Ribobio Co., Ltd., Guangzhou, China), control-shRNA (Santa Cruz Biotechnology, USA), CD99-shRNA (Santa Cruz Biotechnology, USA), or miR-663b inhibitor+CD99-shRNA for 48 h by using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. After transfection, the efficiency of transfection was evaluated by qRT-PCR. Then, MTT assay, trans-well assay, and flow cytometry assay were performed.

Liu X., Zhang H., Zhang B, & Zhang X. (2019). Expression and Role of MicroRNA-663b in Childhood Acute Lymphocytic Leukemia and its Mechanism. Open Medicine, 14, 863-871.

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Regulation of Fibroblast Behavior by Smad7 and miR-497-5p

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Human embryonic skin fibroblasts CCC-ESF-1 (cat. no. YB-ATCC-3084; Shanghai Zibo Biological Technology Co., Ltd.) and human HS fibroblasts (hHSFs; cat. no. C0618; Shanghai Guandao Biological engineering Co., Ltd.) were cultured in RPMI-1640 medium (Gibco; Thermo Fisher Scientific, Inc.) containing 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37˚C with 5% CO₂.
Control-plasmid (1 µg; cat. no. sc-437275; Santa Cruz Biotechnology, Inc.), Smad7-plasmid (1 µg; cat. no. sc-400251-ACT; Santa Cruz Biotechnology, Inc.), miR-497-5p inhibitor (100 nM; 5'-ACAAACCACAGTGTGCTGCTG-3'; Sangon Biotech Co., Ltd.), inhibitor control (100 nM; 5'-CAGTACTTTTGTGTAGTACAA-3'; Sangon Biotech Co., Ltd.), Smad7-short hairpin (sh)RNA (1 µg; cat. no. sc-36508-SH; Santa Cruz Biotechnology, Inc.), control-shRNA (1 µg; cat. no. sc-108060; Santa Cruz Biotechnology, Inc.) or 100 nM miR-497-5p inhibitor + 1 µg Smad7-shRNA were transfected into hHSFs (5x10⁴ per well) using Lipofectamine^® 2000 (Invitrogen; Thermo Fisher Scientific, Inc.) according to the manufacturer's instructions. At 48 h post-transfection, transfection efficiency was assessed via reverse transcription-quantitative PCR (RT-qPCR).

Li Z., Wang P., Zhang J, & Zhao D. (2021). MicroRNA-497-5p downregulation inhibits cell viability, reduces extracellular matrix deposition and induces apoptosis in human hyperplastic scar fibroblasts by regulating Smad7. Experimental and Therapeutic Medicine, 21(4), 384.

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Bcl2 shRNA Lentiviral Knockdown

Cited 1 time

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Bcl2 shRNA and control shRNA were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Hairpin sequence of Bcl2 shRNA: GAT CCG TGT GGA TGA CTG AGT ACC TGA TTC AAG AGA TCA GGG ACT CAG TCA TCC ACA TTT TTG. Hairpin sequence of control shRNA: GAT CCG GAA CGG CAT CAA GGT GAA CTT CAA GA GAG TTC ACC TTG ATG CCG TTC TTT TTG. For pseudovirus production, Bcl2 shRNA or control shRNA was cotransfected into 293FT cells with a lentivirus packaging plasmid mixture (System Biosciences, CA, USA) using the NanoJuice transfection kit (EMD Chemical, Inc.) as described (25 (link)). After 48 h, the virus-containing media were harvested by centrifugation at 20 000 × g. H460 cells were infected with the virus-containing media in the presence of polybrene (8 μg/ml) for 24 h. Stable positive clones were selected using 1 μg/ml puromycin. Speciﬁc silencing of the targeted Bcl2 gene was conﬁrmed by at least three independent experiments.

Xie M., Park D., You S., Li R., Owonikoko T.K., Wang Y., Doetsch P.W, & Deng X. (2015). Bcl2 inhibits recruitment of Mre11 complex to DNA double-strand breaks in response to high-linear energy transfer radiation. Nucleic Acids Research, 43(2), 960-972.

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Romo1 Modulation in Cell Culture

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We obtained RPMI 1640 medium, fetal bovine serum (FBS), and antibiotics from Gibco (Carlsbad, CA, USA), and obtained antibodies including Romo1 (OriGene Technologies, Rockville, USA) and β-actin (Sigma, St. Louis, MO, USA) as well. We obtained Romo1 siRNA, Romo1 shRNA, control siRNA, control shRNA, and plasmid DNA (pFlag-c1 (control) and pFlag-c1 Romo1) from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Kim H.J., Jo M.J., Kim B.R., Kim J.L., Jeong Y.A., Na Y.J., Park S.H., Lee S.Y., Lee D.H., Lee H.S., Kim B.H., Lee S.I., Min B.W., Yoo Y.D, & Oh S.C. (2017). Reactive oxygen species modulator-1 (Romo1) predicts unfavorable prognosis in colorectal cancer patients. PLoS ONE, 12(5), e0176834.

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shRNA Knockdown in mAPSCs

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For shRNA experiments, cells were plated into 10 cm² culture dishes at a density of 5 x 10⁴/cm² and infected with Lenti-FOXC1 shRNA (LV-FOXC1-sh, sc-43766-V, Santa Cruz Biotechnology), Lenti-STI-1 shRNA (LV-STI-1-sh, sc-153893-V, Santa Cruz Biotechnology), Lenti-PrP^C shRNA (LV-PrP^C-sh, sc-36319-V, Santa Cruz Biotechnology) and control shRNA (Santa Cruz Biotechnology) for 48 hours following the manufacturer's instruction. Cell proliferation assays and protein expression of FOXC1, STI-1 and PrP^C, as determined by western blot, were examined for LV-FOXC1-sh-, LV-STI-1-sh- and LV-PrP^C-sh-mAPSCs.

Lee Y.H., Lee H.T., Chen C.L., Chang C.H., Hsu C.Y, & Shyu W.C. (2019). Role of FOXC1 in regulating APSCs self-renewal via STI-1/PrPC signaling. Theranostics, 9(22), 6443-6465.

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Notch Signaling Pathway Regulation

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ZER (purity >98%) was purchased from Sigma-Aldrich (St. Louis, MO). Reagents necessary for cell culture, including fetal bovine serum and antibiotics, and Oligofectamine were purchased from Invitrogen-Life Technologies (Carlsbad, CA). Antibodies for detection of cleaved Notch1, Jagged1, Jagged2, Notch2, cleaved poly-(ADP-ribose)-polymerase (PARP), cleaved caspase-3, Bcl-2, Presenilin-1, and Nicastrin were from Cell Signaling Technology (Beverly, MA); an antibody specific against cleaved Notch2 was from EMD-Millipore (Billerica, MA); anti- Notch4 (detects both full length and cleaved form) antibody was from Santa Cruz Biotechnology (Dallas, TX); an antibody specific for immunodetection of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was from GeneTex (Irvine, CA); and anti-actin antibody was from Sigma-Aldrich. The Notch2 and Presinilin1-targeted small interfering RNA (siRNA), Notch2-targeted small hairpin RNA (shRNA), and control shRNA were purchased from Santa Cruz Biotechnology.. A nonspecific control siRNA was purchased from Qiagen (Germantown, MD). MCF-7 and MDA-MB-231 cells were purchased from American Type Culture Collection (Manassas, VA) and maintained as described previously [27 (link)]. SUM159 cells (Asterand, Detroit, MI) were cultured as suggested by the supplier.

Sehrawat A., Sakao K, & Singh S.V. (2014). Notch2 activation is protective against anticancer effects of zerumbone in human breast cancer cells. Breast cancer research and treatment, 146(3), 543-555.

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MITF Knockdown via shRNA Lentivirus

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Short hairpin RNA (shRNA) targeting the coding sequence of MITF and control shRNA were purchased from Santa Cruz. Lentiviruses encoding control shRNA and MITF shRNA were produced in HEK-293T cells by transient transfection of lentiviral-based vectors and their packaging vectors psPAX2 and pMD2.G. The virus was collected, filtered through a 0.45 µm syringe filter after 48 h, and the M397 cells were spin-infected with viral supernatant supplemented with 10 µg/mL polybrene at 900 g and 30 °C for 90 min. The transduced cells were selected using puromycin, starting at 3 days post transduction.

Su Y., Ko M.E., Cheng H., Zhu R., Xue M., Wang J., Lee J.W., Frankiw L., Xu A., Wong S., Robert L., Takata K., Yuan D., Lu Y., Huang S., Ribas A., Levine R., Nolan G.P., Wei W., Plevritis S.K., Li G., Baltimore D, & Heath J.R. (2020). Multi-omic single-cell snapshots reveal multiple independent trajectories to drug tolerance in a melanoma cell line. Nature Communications, 11, 2345.

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Lentiviral Vector Transfection Protocol

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p21CIP1 expression vector was obtained from Dr. Stuart Aaronson (Mount Sinai School of Medicine, New York, NY, USA). Mouse p21CIP1 shRNA and control shRNA were obtained from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). Mouse LEF1, TCF1, Cyclin D1 shRNA clones and non-silencing control shRNA in the pLKO.1 lentiviral vector (Open Biosystems) used to knock down these molecules were obtained from the genomic core facility at Einstein. To generate viruses, lentiviral vectors were transfected into 293T cells with Tat, Rev, Gag/Pol, and VSV-G vectors. TOPFlash, FOPFlash and Renilla plasmids were obtained from Millipore.

Benard O., Qian X., Liang H., Ren Z., Suyama K., Norton L, & Hazan R.B. (2019). p21CIP1 promotes mammary cancer initiating cells via activation of Wnt/TCF1/CyclinD1 signaling. Molecular cancer research : MCR, 17(7), 1571-1581.

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Genetic Manipulation of Neural Cells

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Control shRNA, parkin shRNA, and p21 shRNA were purchased from Santa Cruz Biotechnology. The viral particles were combined with 8 μg/mL of polybrane (Santa Cruz Biotechnology) and infected overnight into neural stem cells or PC12 cells. The cell culture medium was replaced with fresh complete growth medium and after 48 hours, and the cells were used for experiments.

Park M.H., Lee H.J., Lee H.L., Son D.J., Ju J.H., Hyun B.K., Jung S.H., Song J.K., Lee D.H., Hwang C.J., Han S.B., Kim S, & Hong J.T. (2017). Parkin Knockout Inhibits Neuronal Development via Regulation of Proteasomal Degradation of p21. Theranostics, 7(7), 2033-2045.

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Stable STAT3 Knockdown in Daoy and D556 Cells

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STAT3 shRNA plasmid (h) (#sc‐29493) and control shRNA(#sc‐108080) were purchased from Santa Cruz Biotechnology. Puromycin was used to select Daoy or D556 cells transfected with human STAT3 shRNA or control shRNA. After removing the medium with dead cells under puromycin selection, single colonies were picked up from the culture dishes and then put into single wells in a new 96‐well plate to grow. The cells were kept growing in the same medium with puromycin until enough cells were isolated to confirm the resulting expression of STAT3 by western blot.

Yuan L., Zhang H., Liu J., Malhotra A., Dey A., Yu B., Jella K.K., McSwain L.F., Schniederjan M.J, & MacDonald T.J. (2021). STAT3 is required for Smo‐dependent signaling and mediates Smo‐targeted treatment resistance and tumorigenesis in Shh medulloblastoma. Molecular Oncology, 16(4), 1009-1025.

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