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On targetplus smartpool

Manufactured by Thermo Fisher Scientific
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The ON-TARGETplus SMARTpool is a gene silencing reagent designed to efficiently and specifically target and downregulate gene expression. It consists of a pool of four individual siRNA duplexes that are pre-designed and pre-validated to target a specific gene of interest.

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Lab products found in correlation

Lipofectamine 2000, by Thermo Fisher Scientific (18 mentions) Lipofectamine rnaimax, by Thermo Fisher Scientific (16 mentions) Lipofectamine rnaimax transfection reagent, by Thermo Fisher Scientific (8 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (7 mentions) On targetplus non targeting pool, by Thermo Fisher Scientific (5 mentions) Rnaimax, by Thermo Fisher Scientific (5 mentions) Hiperfect transfection reagent, by Qiagen (4 mentions) Hiperfect, by Qiagen (4 mentions) Amaxa nucleofector, by Lonza (4 mentions) Lipofectamine rnaimax reagent, by Thermo Fisher Scientific (3 mentions) Ingenio electroporation solution, by Mirus Bio (3 mentions) Hiperfect reagent, by Qiagen (3 mentions) Opti mem, by Thermo Fisher Scientific (2 mentions) Rneasy mini kit, by Qiagen (2 mentions) Pcdh cmv mcs ef1 puro, by System Biosciences (2 mentions) Oligofectamine reagent, by Thermo Fisher Scientific (2 mentions) Silencer select, by Thermo Fisher Scientific (2 mentions) Rhs4459, by Thermo Fisher Scientific (2 mentions) Trcn0000151238, by Thermo Fisher Scientific (2 mentions) Lamtor1, by Thermo Fisher Scientific (2 mentions)

Close spelling variants of this product

ON-TARGETplus SMARTpool siRNA (69 citations) SMARTpool (46 citations) ON-TARGETplus (23 citations) ON-TARGETplus SMARTpool human siRNAs (6 citations) SMARTpool: ON-TARGETplus Mcl-1 siRNA, (3 citations) IKZF3 ON-Targetplus siRNA SMARTpools (2 citations) SiRNAs (ON-TARGETplus SMARTpool) (2 citations) ON-TARGETplus SMARTPOOL human CTNNB1 (2 citations) ON-TARGETplus SMARTpool Human PPARG (2 citations) ON-TARGETplus SMARTpool custom siRNA libraries (2 citations)

112 protocols using on targetplus smartpool

1

Par-4 and CK2 Protein Interaction Assay

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GST-Par-4 was a gift from Dr. Robert (University of Manchester); GFP-CK2α and β constructs and their corresponding mutants were previously described.59 (link) The rat constructs pcDNA3.1 TOPO GFP-Par-4 mutants (124A223A, 124D223D, 124A, 124D, 223A, 223D, 123A), the Flag or GFP deletant rat Par-4 (124–332) and the human constructs pcDNA3.1 Myc-Par-4 (wild-type, 231A and 231D mutants) were performed by directed mutagenesis following the manufacturer's instructions (Stratagene, Agilent Technology, Massy, France). Cell lines were transfected for 15 h (no significant apoptosis due to Par-4 overexpression is detected at that time29 (link) and data not shown) using Nanojuice (Millipore, Molsheim, France) or Jetpei (Polyplustransfection, Ozyme, St Quentin en Yvelines, France). PC3 cells were transfected with interferin reagent using two different human CK2 siRNA (siCK2T (L-003475-00-0005) ON-TARGETplus SMARTpool from Thermo Fisher Scientific (Waltham, MA, USA) or siCK2AB from Ambion (Life Technology, Saint Aubin, France, S3638)) and/or two different human Par-4 siRNA (siPar-4T (L-004434-00-0005) ON-TARGETplus SMARTpool from Thermo Fisher Scientific (or siPar-4sc (SC-36190) from Santa Cruz (Tebu-Bio, Le Perray en Yveline, France)) together with scrambled siRNA fluorescently labeled with FAM (Scr siRNA, Santa Cruz) for 48 h.
de Thonel A., Hazoumé A., Kochin V., Isoniemi K., Jego G., Fourmaux E., Hammann A., Mjahed H., Filhol O., Micheau O., Rocchi P., Mezger V., Eriksson J.E., Rangnekar V.M, & Garrido C. (2014). Regulation of the proapoptotic functions of prostate apoptosis response-4 (Par-4) by casein kinase 2 in prostate cancer cells. Cell Death & Disease, 5(1), e1016-.
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2

Silencing HIF-1α and HIF-2α in Macrophages

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For siRNA transfections human macrophages were seeded in 6-well plates as described above. 50 nM siRNA against HIF-1α (ON-TARGETplus SMART pool, Human HIF1A, Thermo Scientific, Karlsruhe, Germany), HIF-2α (ON-TARGETplus SMART pool, Human EPAS1, Thermo Scientific), or non-targeting siRNA pool (GE Healthcare Dharmacon) was used with 16.8 μl HiPerfect (Qiagen, Hilden, Germany) in 500 μl medium with penicillin and streptomycin for each well. After adding the mix, cells were incubated for 24 h. Then medium was removed and replaced by medium containing penicillin, streptomycin and human serum to start experiments.
Cui W., Zhou J., Dehne N, & Brüne B. (2015). Hypoxia induces calpain activity and degrades SMAD2 to attenuate TGFβ signaling in macrophages. Cell & Bioscience, 5, 36.
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3

Transient Knockdown of SMAD4 and Furin

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We performed transient knockdown assays using 25 or 50nM SMAD4-targeting small interfering RNA (siRNA; ON-TARGETplus SMARTpool), 25nM furin-targeting siRNA (ON-TARGETplus SMARTpool) or 25 or 50nM control siRNA (ON-TARGETplus Nontargeting Pool) purchased from ThermoFisher Scientific. Cells were precultured in antibiotic-free DMEM/F12 medium containing 10% fetal bovine serum until they were 50 -60% confluent and then transfected with siRNA using Lipofectamine RNAiMAX (Life Technologies) for 48 hours. Knockdown efficiency for each target was confirmed by RT-qPCR and/or Western blot.
Chang H.M., Cheng J.C., Klausen C, & Leung P.C. (2015). Recombinant BMP4 and BMP7 increase activin A production by up-regulating inhibin βA subunit and furin expression in human granulosa-lutein cells. The Journal of clinical endocrinology and metabolism, 100(3).
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4

RNF2 and Bmi1 siRNA Knockdown

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Dharmacon on-TARGETplus Smartpool siRNA to RNF2 and Bmi1 were delivered using Lipofectamine RNAiMAX (Invitrogen). Non-target Pool siRNA was used as control.
Zhang Y., Li X., Chen Z, & Bepler G. (2014). Ubiquitination and Degradation of Ribonucleotide Reductase M1 by the Polycomb Group Proteins RNF2 and Bmi1 and Cellular Response to Gemcitabine. PLoS ONE, 9(3), e91186.
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5

Silencing FABP7 and FABP6 in SKRC Cells

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A commercially available mixture of 4 single-stranded 19-bp siRNAs (ON-TARGETplus SMARTpool, Invitrogen®) was used to transfect SKRC7 and SKRC10 cells according to the manufacturer’s instructions. The sequences of the siRNAs were 5’CAACGGUAAUUAUCAGUCA3’, 5’GUCAGAACUUUGAUGAGUA3’, 5’GAACACGGAGAUUAGUUUC3’, and 5’GAUGAUAGAAACUGUAAGU3’ for FABP7, 5’UCGGAGGCGUGACCUAUGA3’, 5’CCUCAGAGAUCGUGGGUGA3’, 5’GUGAGAAGAAUUAUGAUGA3’, and 5’GCAAGGAAAGCAACAUACA3’ for FABP6.
The final concentration of siRNA for transfection was 33 nmol/L. A scrambled siRNA served as a negative control (Invitrogen®). Transfection was carried out using Lipofectamine 2000 (Invitrogen®).
Nagao K., Shinohara N., Smit F., de Weijert M., Jannink S., Owada Y., Mulders P., Oosterwijk E, & Matsuyama H. (2018). Fatty acid binding protein 7 may be a marker and therapeutic targets in clear cell renal cell carcinoma. BMC Cancer, 18, 1114.
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6

Astrocyte-Neuron Communication in Ataxin-1 Pathology

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C8S (ATCC, CRL-2535, Astrocyte type II clone) cells were plated at 2 × 105 cells per 12-well plate 24 h prior to transfection, maintained in a 37 °C, 5.5% CO2 incubator. When 70 to 80% confluent, C8S cells were transfected with 1 μg of Flag-hATXN1 [82Q] plasmid, Lipofectamine 3000 (Invitrogen, L3000015), P-3000 (Invitrogen, L3000015), and Opti-MEM. Following an 8-h transfection period, media was changed and C8S cells were transfected with 20 nM control siRNA (Dharmacon, ON-TARGETplus nontargeting control pool, D-001810-10-20) or Ctnnb1 siRNA (Dharmacon, ON-TARGETplus SMARTpool, L040628-00-0005), Lipofectamine 3000 (Invitrogen, L3000015), and Opti-MEM for 12 h. ACM was transferred to untreated N2A (ATCC, CCL-131) cells the following day. N2A cells had been plated at 2 × 105 cells per 12-well plate 24 h prior to addition of ACM and maintained in a 37 °C, 5.5% CO2 incubator. C8S cells were subsequently collected and transferred into 500 μL PBS and centrifuged for 3 min at 10,000 rpm. RNA was extracted from pelleted C8S cells using Qiagen RNeasy Mini Kit, following the manufacturer’s instructions. N2A cells were cultured with ACM for 24 h, and subsequently collected and transferred into 500 μL PBS and centrifuged for 3 min at 10,000 rpm. Protein was extracted from pelleted N2A cells and used for Western blot analysis.
Luttik K., Tejwani L., Ju H., Driessen T., Smeets C.J., Edamakanti C.R., Khan A., Yun J., Opal P, & Lim J. (2022). Differential effects of Wnt-β-catenin signaling in Purkinje cells and Bergmann glia in spinocerebellar ataxia type 1. Proceedings of the National Academy of Sciences of the United States of America, 119(34), e2208513119.
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7

Modulating Target Gene Expression in WJ-MSCs

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For short interfering RNA (siRNA) studies, WJ-MSC lines were transfected with 100nM siRNA (Dharmacon ON-TARGETplus SMARTpool) twice at 24h intervals, using Lipofectamine™ RNAiMAX Transfection Reagent (Invitrogen, 13778150) according to the manufacturer’s protocol. The siRNA sequences are listed in S1 Table. For overexpression studies, cells were transfected with 10 μg of DDK-tagged plasmid DNA from Origene (pCMV6 empty vector, PS100001 or pCMV6-ELOVL2, RC209232) using Lipofectamine® LTX with Plus™ Reagent (Invitrogen, 15338100) according to the manufacturer’s protocol. Cells were harvested or assessed in downstream assays after 48 hrs from the second round of siRNA transfection or after 72 hrs from plasmid overexpression.
Tan P.Y., Chang C.W., Duan K., Poidinger M., Ng K.L., Chong Y.S., Gluckman P.D, & Stünkel W. (2016). E2F1 Orchestrates Transcriptomics and Oxidative Metabolism in Wharton’s Jelly-Derived Mesenchymal Stem Cells from Growth-Restricted Infants. PLoS ONE, 11(9), e0163035.
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8

Exploring Cellular Pathways with siRNA

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Ultrapure LPS (Escherichia coli 0111:B4) was purchased from List Biological Laboratories (Vandell Way, CA). siRNA knockdown was accomplished by transfecting plated cells with siRNA for p62, Nrf2, beclin1, MyD88, and TIRAP (Thermo Scientific ON-TARGET plus SMARTpool) with Lipofectamine 2000 (Invitrogen) in OPTI-MEM media (Gibco by Life Technologies) for at least 24 h. Cell media was changed to antibiotic-free media prior to adding siRNA. EBSS media were purchased from Invitrogen.
Chen C., Deng M., Sun Q., Loughran P., Billiar T.R, & Scott M.J. (2014). Lipopolysaccharide Stimulates p62-Dependent Autophagy-Like Aggregate Clearance in Hepatocytes. BioMed Research International, 2014, 267350.
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9

siRNA-Mediated Gene Knockdown Prior to CRISPR

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All siRNAs were obtained from Dharmacon as ON-TARGETplus Smartpool and transfected on 1 million cells with the RNAiMAX Transfection kit (Invitrogen) at a final concentration of 25 nM, 24h prior to sgRNA electroporation according to the manufacturer’s protocol for 6 well plates. Samples were partially collected 24h after electroporation for subsequent RNA isolation, reverse transcription and qPCR analysis. The rest was left to grow for 48 more hours CRISPR/Cas9 editing analysis.
Schep R., Brinkman E.K., Leemans C., Vergara X., van der Weide R.H., Morris B., van Schaik T., Manzo S.G., Peric-Hupkes D., van den Berg J., Beijersbergen R.L., Medema R.H, & van Steensel B. (2021). Impact of chromatin context on Cas9-induced DNA double-strand break repair pathway balance. Molecular Cell, 81(10), 2216-2230.e10.
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10

Efficient GDA Knockdown Using siRNA

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GDA knockdown was achieved after the second UVB irradiation or the day after cell seeding by transfecting the cells with 500 nM siRNA against human GDA or a negative control (On-TARGETplus SMARTpool or Non-targeting siRNA; Invitrogen) using the TransIT-siQUEST transfection reagent (Mirus Bio, Madison, WI, USA) according to the manufacturer’s protocol.
CHEONG K.A, & LEE A.Y. (2020). Guanine Deaminase Stimulates Ultraviolet-induced Keratinocyte Senescence in Seborrhoeic Keratosis via Guanine Metabolites. Acta Dermato-Venereologica, 100(8), 5732.
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