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Short hairpin rna

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Short hairpin RNA (shRNA) is a type of laboratory equipment used in molecular biology experiments. It consists of a short, double-stranded RNA molecule designed to target and silence specific genes. The core function of shRNA is to induce RNA interference, a biological process that can selectively inhibit the expression of target genes.

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Lab products found in correlation

Fm4 64, by Thermo Fisher Scientific (3 mentions) Shrna construct, by Merck Group (2 mentions) Thapsigargin, by Enzo Life Sciences (1 mentions) Rapamycin, by Thermo Fisher Scientific (1 mentions) Dynasore, by Merck Group (1 mentions) Lucifer yellow, by Thermo Fisher Scientific (1 mentions) Cycloheximide (chx), by Avantor (1 mentions) Mg132, by Avantor (1 mentions) Gibson assembly cloning kit, by New England Biolabs (1 mentions)

Close spelling variants of this product

Mission short hairpin RNA (shRNA) lentiviral vectors (4 citations) MISSION short-hairpin RNA (shRNA) plasmids (3 citations) Lentiviral short hairpin RNA (shRNA) constructs (3 citations) Short-hairpin RNA (shRNA) (3 citations) Lentiviral short hairpin RNAs targeting (2 citations) Lentiviral short hairpin (sh) RNAs (2 citations) Mission short-hairpin RNAs (shRNAs) (2 citations) Short hairpin RNAs (shRNAs) (2 citations) Lentiviral short hairpin RNA (shRNA) transduction particles (2 citations) Lentiviral Mission short hairpin RNA (shRNA) clones targeting (2 citations)

6 protocols using short hairpin rna

1

Lentiviral Knockdown of MUC1 Gene

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For the generation of MUC1 knockdown cells, short hairpin RNA (shRNA) constructs, with scrambled SCR or targeting two independent regions of MUC1 mRNA, were obtained from Millipore Sigma (Burlington, MA USA). These lentiviral constructs were used to generate packaged lentiviruses by transfecting the constructs with packaging constructs into HEK293T cell to produce supernatants which were used to transduce indicated cells. Cells were selected at 48 hours post infection using puromycin for 72 hours or until complete death of un-infected cells.
Olou A.A., King R.J., Yu F, & Singh P.K. (2020). MUC1 Oncoprotein Mitigates ER Stress via CDA-mediated Reprogramming of Pyrimidine Metabolism. Oncogene, 39(16), 3381-3395.
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2

Lentiviral Knockdown of MUC1 Gene

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For the generation of MUC1 knockdown cells, short hairpin RNA (shRNA) constructs, with scrambled SCR or targeting two independent regions of MUC1 mRNA, were obtained from Millipore Sigma (Burlington, MA USA). These lentiviral constructs were used to generate packaged lentiviruses by transfecting the constructs with packaging constructs into HEK293T cell to produce supernatants which were used to transduce indicated cells. Cells were selected at 48 hours post infection using puromycin for 72 hours or until complete death of un-infected cells.
Olou A.A., King R.J., Yu F, & Singh P.K. (2020). MUC1 Oncoprotein Mitigates ER Stress via CDA-mediated Reprogramming of Pyrimidine Metabolism. Oncogene, 39(16), 3381-3395.
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3

Targeted silencing of immune sensors

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Short hairpin RNA (shRNA) against human DAI, LRRFIP1, AIM2, ATG5, TLR4, and RAGE were obtained from Sigma (St. Louis, MO, USA) and were transfected into cells using lentiviral-mediated transfection (Sigma) according to the manufacturer’s instructions. Detection of knockdown was assayed by Western blot.
Liu L., Yang M., Kang R., Yu Y., Dai Y., Gao F., Wang H., Sun X., Li X., Li J., Wang H., Cao L, & Tang D. (2014). HMGB1-DNA Complex-induced Autophagy Limits AIM2 Inflammasome Activation through RAGE. Biochemical and biophysical research communications, 450(1), 851-856.
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4

RNAi Mediated Depletion of HOXB9 and FOXA2

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RNA interference was performed as previously described (Ma et al., 2017; Yang et al., 2015) . The short hairpin RNA was purchased from Sigma-Aldrich (St. Louis, MO). The sequences targeting HOXB9 were 5-CCCTTCAATTTGTAGACTCTT-3 and 5-CTCCTCAATCTGAGTGAGAGA-3 and targeting FOXA2 were 5-CAAGGGAGAAGAAATCCATA-3 and 5-GAACGGCATGAACACGTACAT-3.
Lin J., Zhang D., Fan Y., Chao Y., Chang J., Li N., Han L, & Han C. (2018). Regulation of Cancer Stem Cell Self-Renewal by HOXB9 Antagonizes Endoplasmic Reticulum Stress-Induced Melanoma Cell Apoptosis via the miR-765-FOXA2 Axis. The Journal of investigative dermatology, 138(7).
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5

Plasmid Generation and Reagent Use

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Details of all yeast and human cell line plasmids are listed in Supplementary Data 1. All plasmids generated in this study were constructed using the Gibson assembly cloning kit (NEB), in accordance with manufacturer guidelines. About 30 bp overlaps were utilized for DNA fragment joining; Sanger sequencing confirmed successful cloning. Chemicals used in this paper: FM4-64 (Thermo Fisher Scientific, T3166), Lucifer yellow (Thermo Fisher Scientific, L453), Thapsigargin (Enzo Life Sciences, BML-PE180-0001), Dynasore (Sigma Aldrich, D7693), MG132 (VWR, 89161-566), CHX (Amresco, 94271-5G), Rapamycin (Fisher Scientific, BP2963-1). ATG5 siRNA was purchased from CST. Short hairpin RNAs were bought from Sigma Aldrich.
Liu G., Coyne A.N., Pei F., Vaughan S., Chaung M., Zarnescu D.C, & Buchan J.R. (2017). Endocytosis regulates TDP-43 toxicity and turnover. Nature Communications, 8, 2092.
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6

Lentiviral shRNA Transfection in 293T, Kasumi-1, and K562 Cells

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Human embryonic kidney 293T cells (ATCC) were cultured in Dulbecco's modified Eagle's medium supplemented with streptomycin (100 µg/ml), penicillin (50 U/ml), and 10% fetal bovine serum. Short hairpin RNAs (shRNA; Sigma-Aldrich) were cotransfected into 293T cells along with pHR and VSVG plasmids to produce shRNA lentiviruses. Kasumi-1 and K562 cells were transfected with shRNA lentiviruses targeting GNAS (shGNAS) or a negative control (shNC).
He S., Li Y., Wang L., Li Y., Xu L., Cai D., Zhou J, & Yu L. (2024). DNA methylation landscape reveals GNAS as a decitabine-responsive marker in patients with acute myeloid leukemia. Neoplasia (New York, N.Y.), 49, 100965.
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