The siRNA sequence 5′-AAAGCCUUCUGUGCUGAGAAC-3′ was used to target mouse β-arrestin 1 (position 439–459 relative to the start codon) whereas sequence 5′-AAACCUGUGCCUUCCGCUAUG-3′ was used to target mouse β-arrestin 2 (position 175–193 relative to the start codon)39 (link). One small RNA duplex with no silencing effect was used as a control (5′-UUCUCCGAACGUGUCACGU-3′). The siRNAs were synthesized by GE Healthcare Dharmacon (Velizy-Villacoublay, France). Early passage mLTC-1 cells at 30% confluency in 100 mm dishes were transiently transfected with GeneSilencer following the manufacturer’s recommendations (Genlantis, San Diego, CA, USA). Forty-eight hours after transfection, cells were seeded into assay plates. All assays were performed three days after transfection.
Genesilencer
Manufactured by Genlantis
Sourced in United States
GeneSilencer is a laboratory equipment product that is used for the delivery of small interfering RNA (siRNA) or other nucleic acid molecules into cells for the purpose of gene silencing. The core function of GeneSilencer is to facilitate the introduction of these molecules into target cells, enabling the downregulation or suppression of specific gene expression.
Automatically generated - may contain errors
Lab products found in correlation
Sigenome smartpool, by Horizon Discovery (1 mentions)
Lipopolysaccharide lps, by Merck Group (1 mentions)
Sc 35674, by Santa Cruz Biotechnology (1 mentions)
On targetplus non targeting pool, by Horizon Discovery (1 mentions)
Siglo red transfection indicator, by Horizon Discovery (1 mentions)
Smartpool, by Horizon Discovery (1 mentions)
On targetplus, by Horizon Discovery (1 mentions)
Mirvana mirna mimic, by Thermo Fisher Scientific (1 mentions)
Block it rnai designer, by Thermo Fisher Scientific (1 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
14 protocols using genesilencer
1
Targeting beta-arrestin with siRNA
2
Silencing STIM1 and Orai1 in cells
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Small interfering RNA (siRNA) targeted to STIM1 (siSTIM1) and Orai1 (siOrail1) were designed and synthesized by Dharmacon (accession numbers XM_341896; siGENOME SMARTpool; Dharmacon, Lafayette, CO) and Shanghai GenePharma respectively. Primary cultured cells at 50–70% confluence were transfected with siSTIM1 or siOrai1, and nontargeting control siRNA using transfection vehicle (GeneSilencer; Genlantis, San Diego, CA) as carrier for 4 hours in serum-free SMBM in 5% CO2 at 37°C. The final concentration of each siRNA was 1000 ng/ml.
3
Transient miR-223-3p Overexpression in BMDCs
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For transient overexpression of mmu-miR-223-3p, BMDCs were harvested on d6 of culture and reseeded (106) onto polycarbonate coated transwell inserts (0.8 μm diameter; Corning Costar, Corning, NY) placed into untreated 6 well culture plates (Greiner Bio-One). Then, BMDC culture medium was exchanged against serum-free medium (2 ml). BMDCs were transfected in parallel with each 1 nmol of mmu-miR-223-3p mimick (sense: 5′-UGUCAGUUUGUCAAAUACCCCA-3′; Qiagen, Hilden, Germany) and FITC-labeled scrambled control siRNA (sense: 5′-AGGUCGAACUACGGGUCAAUC-3′; NEB, Ispwich, MA) complexed with GeneSilencer (Genlantis, San Diego, CA). After 4h, medium was exchanged against BMDC culture medium. On the next day, BMDCs were transfected again. On d8, aliquots of transfectants were stimulated with LPS (1 μg/ml) overnight. On the following day, BMDCs were thoroughly washed and used for experiments.
4
Silencing BMPRII in PASMCs
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BMPRII small interference RNA (siRNA accession no. XM_217409) were designed and synthesized by Dharmacon (Lafayette, CO). Nontargeting siRNA served as a control. Transfection reagent obtained from GeneSilencer (Genlantis) was used according to the manufacturer's instructions. PASMCs at 50–60% confluence were transfected with 50 nM siRNA for 6 h in serum and antibiotic-free SMBM, then cultured for 42 h in SMBM with 0.3% FBS, followed by treatment with BMP4 at different time point.
5
Generating Bone Marrow-derived DCs
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Bone marrow-derived DCs (BMDCs) were obtained as previously described [10 (link)], washed with a serum-free medium, and cultured in a 24-well tissue culture plate with 2 × 105 cells/500 μl. Transfection reagent (Gene Silencer®, Genlantis) and siRNA were added to the wells. After 4 hours, 500 μl medium with 20% FBS was added. After 24 hours, lipopolysaccharide (LPS) (1 μg/ml; Sigma, St. Louis, MO, USA) and peptide (p) OVA (C-terminal 323–339 epitope) (5 μg/ml; Bachem, Hauptstrasse, Switzerland) were added for 18 hours.
6
β-arrestin-2 Knockdown in HEK293T Cells
7
Modulating Cell Adhesion via β1 Integrin and Galectin-3
8
Silencing PRKG1 in Young VSMC
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VSMC from young (3 months old) rats were used for silencing of PKG1 gene. Double‐stranded siRNAs were used for the silencing of PRKG1 (L‐085799‐01‐0050; former Dharmacon Inc., present Horizon Discovery Ltd.); ON‐TARGETplus Non‐targeting Pool (D‐001810‐10‐20; Dharmacon Inc.) was used as a negative control. The transfection of VSMC was conducted with 100 nmol/L siRNA using Gene Silencer as a transfection reagent (T500750; Genlantis Inc., San Diego, CA) according to the manufacturer's instructions. Transfection was made during 6 hours in DMEM; after 6 hours incubation FBS was added to the final concentration 10%; medium was completely changed after 24 hours of incubation with siRNA to DMEM with 10% FBS and antibiotics. Cells were collected for the Western blot analysis after the inclubation with 1 nmol/L marinobufagenin or α‐hANP 1 nmol/L or combination of both compounds following 96 hours after transfection with si‐PKG1.
9
Cochlear Stem Cell Differentiation
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Cochlear neurospheres or organ of Corti explants were transfected with siRNA (100 nM ON-TARGET plus, Smart Pool from Dharmacon for Sox2, or non-targeting siRNA), using Gene Silencer (Genlantis) for 24 hours according to the manufacturer’s instructions. Transfection efficiency was monitored using siGlo Red Transfection Indicator (Dharmacon). In addition, 2.5 μM DAPT was added during differentiation culture. After transfection, the cells were washed and cultured in DMEM/F12 with DAPT (1:1) and N2/B27 (spheres and explants) or DMEM/10% FBS and DAPT (IEC6 cells and OC-1 cells). For quantitative PCR experiments after siRNA treatment, cells were differentiated for additional 24 hours after transfection. Organ of Corti explants were kept in culture for an additional 24 hours (48 hours total) and differentiating neurospheres were cultured for up to 6 days after siRNA treatment.
10
Lentiviral Gene Overexpression and siRNA-Mediated Silencing in RPASMCs
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Gene overexpression in RPASMCs was performed via lentivirus-mediated approaches. Chemical synthesized siRNAs (Shanghai GenePharma Co.) were used to silence EP300 expression in primary RPASMCs or HPASMCs using GeneSilencer (Genlantis) as transfection reagent. Two different siRNA sequences targeting rat EP300 were used. Then qRT-PCR was performed 48 h later to analyze the expression level of EP300 in RPASMCs and HPASMCs. The β-actin was used as endogenous control.
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