Luciferase sirna

Manufactured by Thermo Fisher Scientific

Luciferase siRNA is a laboratory reagent used to silence the expression of the luciferase gene in cells. It is a small interfering RNA (siRNA) designed to target and degrade the mRNA transcript of the luciferase gene, effectively reducing the production of the luciferase protein.

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SiRNAs (258 citations) Silencer select siRNAs against luciferase or Esrp2 (2 citations) Stealth RNAi siRNA Luciferase Reporter Control (2 citations)

5 protocols using luciferase sirna

Lipid Nanoparticle Formulation and Characterization

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2-(aminomethyl)butanedioic acid hydrochloride was purchased from Enamine (Monmouth Jct., NJ). Dimethyl Formamide (DMF), Di-tert-butyl dicarbonate (BOC anhydride), Triethylamine (TEA), 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxide hexafluorophosphate (HATU), N,N-Diisopropylethylamine (DIPEA), trifluoroacetic acid (TFA) N,N’-Di-Boc-L-histidine dicyclohexylammonium salt, Nalpha,Nepsilon-Di-Boc-L-lysine and cholesterol were purchased from Fischer Scientific (Pittsburg, PA). 1,2-Distearoyl-sn-glycero-3-phosphoethanolamine-poly(ethylene glycol) (DSPE-mPEG(2000)) was purchased from Nanocs Inc (New York, NY). 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) was purchased from Avanti Polar Lipids. Luciferase siRNA (5’-CUUACGCUGAGUACUUCGAtt-3’), human IKKα siRNA (5′-GGAGAUCUCCGAAAGCUGCtt-3′), human IKBKE siRNA (5′-GGUCUUCAACACUACCAGCtt-3′), mouse IKBKE siRNA (5’-GGUCUUCAACUCAGCCAGCtt-3) and Lipofectamine^® 2000 were ordered from Invitrogen (Carlsbad, CA). 2-(p-toluidino)-6-naphthalene sulfonic acid (TNS) was obtained from Sigma Aldrich. CellTiter-Glo luminescent cell viability assay kit and ONE-Glo^™ luciferase assay system were purchased from Promega (Madison, WI)

Palacios J.L., Zaror I., Martínez P., Uribe F., Opazo P., Socías T., Gidekel M, & Venegas A. (2001). Subset of Hybrid Eukaryotic Proteins Is Exported by the Type I Secretion System of Erwinia chrysanthemi. Journal of Bacteriology, 183(4), 1346-1358.

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Plasmid Construction and siRNA Techniques

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For plasmids, pcDNA‐v5‐WBP2 and pGEX4T1‐GST‐WBP2 were constructed in our laboratory as previously described [17 , 22 (link)]. pRL‐TK (Renilla) was purchased from Promega (Madison, WI, USA). pcDNA3 was purchased from Invitrogen (Carlsbad, CA, USA). NF‐κB luciferase reporter plasmid, pHAGE NF‐κB‐TA‐LUC‐UBC‐GFP‐W, was a gift from D. Kotton (Addgene plasmid #4934; Addgene, Watertown, MA, USA). pCMV4‐HA‐IκBα was a gift from W. Greene (Addgene plasmid # 21985). pCMV‐8xHis‐Ub was a gift from W. Kaelin (Addgene plasmid #107392). pcDNA3‐Flag‐BTRC was a gift from P. Howley (Addgene plasmid #10865). For siRNAs, WBP2 siRNAs and luciferase siRNA were performed from Invitrogen. IκBα and BTRC siRNAs were purchased from Integrated DNA Technologies. The siRNA sequences are listed in Table S1.

Lim Y.X., Lin H., Chu T, & Lim Y.P. (2021). WBP2 promotes BTRC mRNA stability to drive migration and invasion in triple‐negative breast cancer via NF‐κB activation. Molecular Oncology, 16(2), 422-446.

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Luciferase and PCPB2 siRNA Knockdown

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Luciferase siRNA, PCPB2 siRNA (Sense strand 5′-GUCAGUGUGGCUCUCUUAC-3′), scrambled siRNA, and Lipofectamine 2000 were purchased from Invitrogen (Carlsbad, CA). Cell culture reagents were ordered from Mediatech, Inc. (Manassas, VA). PNAs and PNA conjugates were ordered from PNA Bio Inc. (Newbury Park, CA).

Jin W., Jain A., Liu H., Zhao Z, & Cheng K. (2018). Noncovalent Attachment of Chemical Moieties to siRNAs Using Peptide Nucleic Acid as a Complementary Linker. ACS applied bio materials, 1(3), 643-651.

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Hypoxia Mimicry and HIF-2α Silencing Enhance Chemotherapy

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For experiments mimicking hypoxic conditions, cells were cultured with 100 μM deferoxamine mesylate (DFX) (Sigma-Aldrich), a hypoxia mimetic^{[21 (link),22 (link)]}. A Silencer Select short interfering RNA (siRNA) against human HIF-2α (Dharmacon, Lafayette, CO) was delivered to cells using two different transfection systems: (1) Lipofectamine RNAiMAX as described by the manufacturer (Invitrogen); and (2) cationic amphiphilic micelles complexed with transfection lipids as described in a previous publication^{[23 (link)]}. Silencer Select Negative Control siRNA (Invitrogen) or Luciferase siRNA (Invitrogen) was used as a control, nontargeting sequence. After 24 h, 1 mM temozolomide (Invitrogen) was added to the media. After 48 h, an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (Promega, Madison, WI) was performed to evaluate viability as a measure of chemotherapeutic response to a combined HIF-2α siRNA and temozolomide therapy.

Nusblat L.M., Tanna S, & Roth C.M. (2020). Gene silencing of HIF-2α disrupts glioblastoma stem cell phenotype. Cancer Drug Resistance, 3(2), 199-208.

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Hypoxia Mimetic and HIF-2α Knockdown Enhance Temozolomide Sensitivity

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For experiments mimicking hypoxic conditions, cells were cultured with 100 μM deferoxamine mesylate (DFX) (Sigma-Aldrich), a hypoxia mimetic^{[21 (link),22 (link)]}. A Silencer Select short interfering RNA (siRNA) against human HIF-2a (Dharmacon, Lafayette, CO) was delivered to cells using two different transfection systems: (1) Lipofectamine RNAiMAX as described by the manufacturer (Invitrogen); and (2) cationic amphiphilic micelles complexed with transfection lipids as described in a previous publication^{[23 (link)]}. Silencer Select Negative Control siRNA (Invitrogen) or Luciferase siRNA (Invitrogen) was used as a control, nontargeting sequence. After 24 h, 1 mM temozolomide (Invitrogen) was added to the media. After 48 h, an 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay (Promega, Madison, WI) was performed to evaluate viability as a measure of chemotherapeutic response to a combined HIF-2a siRNA and temozolomide therapy.

Nusblat L.M., Tanna S, & Roth C.M. (2020). Gene silencing of HIF-2α disrupts glioblastoma stem cell phenotype. Cancer Drug Resistance, 3(2), 199-208.

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