Regarding the knock down of PSAT1, two human PSAT1 targeted RNAi (RNAi#1: TTCCAAGTTTGGTGTGATT; RNAi#2: ACTCAGTGTTGTTAGAGAT) sequences were obtained from GeneChem Co. Ltd. (Shanghai, China). As a control, scrambled versions of these sequences were used. The sequences shown above were inserted into the GV248 vector plasmid. For the overexpression of PSAT1, full-length human PSAT1 cDNA was cloned into the pLVX-puro vector. Lentiviral particles were constructed and packaged by Shanghai GeneChem Co. Ltd. Briefly, the cells were infected with lentivirus to generate stable cell lines. After 24 h, the cells were transferred to medium containing 4 μg/ml puromycin and were cultured for 3 days.
Lentiviral particles
Manufactured by Genechem
Sourced in China
Lentiviral particles are a type of viral vector used in gene delivery applications. They are capable of transducing a wide range of cell types, including non-dividing and difficult-to-transfect cells. Lentiviral particles can be used to stably integrate genetic material into the host cell's genome.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 3000, by Thermo Fisher Scientific (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Polybrene, by Genechem (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Puromycin, by Thermo Fisher Scientific (2 mentions)
Puromycin, by Merck Group (2 mentions)
Polybrene, by Merck Group (2 mentions)
Puromycin, by Genechem (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Sirna, by Santa Cruz Biotechnology (1 mentions)
Caco 2, by American Type Culture Collection (1 mentions)
Hyclone, by Cytiva (1 mentions)
Hcoepic, by American Type Culture Collection (1 mentions)
Puromycin, by MedChemExpress (1 mentions)
Lentiviral particles, by GenePharma (1 mentions)
Gv358, by Genechem (1 mentions)
Opti medium, by Thermo Fisher Scientific (1 mentions)
Sirna, by Genechem (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
Hct116, by Genechem (1 mentions)
52 protocols using lentiviral particles
1
Knockdown and Overexpression of PSAT1
2
Lentiviral Knockdown of Fos in HT22 Cells
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Lentiviral particles expressing shRNA against Fos and the corresponding control sequence were constructed by Genechem (Shanghai Genechem Co., Ltd.). Knocked-down cells (KD) and negative control cells (NC) were generated by transfecting HT22 cells with pre-synthesized Fos (or control) shRNA Lentiviral particles as per the standard protocol of the manufacturer. At 72-h post-infection, transfected cells emitted a green fluorescence signal, and puromycin (2 μg/ml) was added to the culture medium for selection and further characterization. The target sequences of shRNA of Fos were as follows: KD shRNA: ccGTCTCTAGTGCCAACTTTA. NC shRNA: TTCTCCGAACGTGTCACGT.
3
Lentiviral Transduction of PD-L1 in K7M2 Cells
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Lentiviral particles in the absence or presence of the PD-L1 gene were obtained from Shanghai GeneChem Co., Ltd. (Shanghai, China). The viruses were used to infect cells in the presence of polybrene and enhanced infection solution (Shanghai GeneChem Co., Ltd.). To obtain stable PD-L1 clones from the K7M2 cell line, the green fluorescent expression was observed in infected cells under a fluorescent microscope.
4
Knockdown of DNASE1L3 and RELB in Liver Cells
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Small interfering RNAs (siRNA) against DNASE1L3 or RELB were synthesised by Santa Cruz Biotechnology, Inc. (TX, USA). SMMC7721 cells or LO2 hepatocytes were seeded into 6‐well plates at a density of 2 × 105 cells/well and cultured overnight. The constructed siRNA‐DNASE1L3, siRNA‐RelB or siRNA‐control was transferred into the cells using Lipofectamine 2000 (Invitrogen). To raise transfection efficiency, the cells were incubated with 20 nm siRNA for 6 h followed by a 24‐h culture. In mouse HCC H22 cells, lentivirus‐mediated shRNA delivery system was used to knock down DNASE1L3 gene. Briefly, H22 cells were seeded into 6‐well plates at a density of 2 × 105 cells/well to culture overnight, and then, the medium containing the lentiviral particles (GeneChem Co., Ltd, Shanghai, China) was supplemented. After a 12‐h infection, the viral medium was replaced with a fresh medium followed by a 72‐h growth. Subsequently, the cells were selected with 1.5 μg mL−1 puromycin for 48 h. The interference efficiency was evaluated by Western blot assay.
5
Overexpression of hsa-miR-374a in NSCLC
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Lentiviral particles carrying hsa-miR-374a precursor and its flanking control sequence (NC for short) were constructed by GeneChem, Shanghai, China. A549, pc-9, SPCA-1, and H1975 cells were infected with lentiviral vector and polyclonal cells with green fluorescent protein signals were selected for further experiments using fluorescence-activated cell sorting flow cytometry. Total RNA from these cell clones was isolated, and levels of miR-374a were quantified using qRT-PCR.
6
Lentiviral Transduction of Osteosarcoma Cells
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Lentiviral particles and scrambled control Lentiviral particles were provided by Shanghai Genechem Co., Ltd., and 1 × 105 osteosarcoma cells were seeded into 6‐well culture plates and infected with Lentiviral particles at 50% confluence with a multiplicity of infection of 50. The infected cells were subsequently selected in the presence of 4 µg/mL puromycin (Thermo Fisher Scientific, USA) for 7 days and maintained in medium containing 2 µg/mL puromycin to obtain stably infected cells. Real‐time q‐PCR and western blotting were performed to analyze the effect of viral infection.
7
Notch Signaling Modulation in HCC Cells
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For suppression or inhibition of Notch in HCC cells, lentiviral particles (Genechem, Shanghai) expressing Notch1-siRNA or NICD were used to regulate Notch signaling in sorted HCC cells. The siRNA sequence targeted Notch1 was listed as follows: 5′-GGAGCATGTGTAACATCAA-3′. For optimization of transfection conditions with lentiviral vectors, HCC cells were infected with Lv-NICD or Lv-Notch1-si at different of multiplicity of infection (MOI) for 12 hours in the presence of 5 μg/ml of polybrene. Two days after infection, expression of green florescence protein was measured by FACS analysis. Infection of cells at MOI of 10 resulted in more than 90% of efficiency of infection without damaging cells (not shown in data).
8
Overexpression of miR-4310 via Lentivirus
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Lentiviral particles encoding hsa-miR-4310 were designed and constructed by GeneChem (Shanghai, China). Cells were infected with lentiviral vector, and the expression of miR-4310 was detected by qPCR.
9
Modulating tsRNA-16902 and RARγ in hMSCs
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Knockdown of tsRNA-16902 and overexpression of RARγ was conducted using lentiviral particles from Shanghai Genechem Co., Ltd. A short hairpin RNA (shRNA) targeting tsRNA-16902 was designed (target sequence,5′-TGGTGTCCTTGGAAAAAGGTTTTCATCTCCGGTTTACAA-3′). In addition, a negative control (NC) shRNA was used (sequence, 5′-TTCTCCGAACGTGTCACGT-3′). Lentiviral transduction reactions were conducted by plating 5×10 4 hMSCs/cm 2 in 6-well plates until 20-30% con uent, after which they were infected using 10 uL of the appropriate lentivirus (1×10 8 infectious units/ml) in complete media supplemented with 5 µg/ml polybrene. After 10 h, this transduction media was removed and fresh media was added, after which the cells were grown for an additional 72 h. Cells were then cultured in 0.5µg/ml puromycin for 48 h, after which they were screened over a 6 day period with fresh media added every 1-2 days.
At various time points during adipogenesis, Oil Red O staining was used to con rm the presence of lipid droplets within cells. In addition, cells were collected to assess the expression of adipogenic markers and RARγ.
At various time points during adipogenesis, Oil Red O staining was used to con rm the presence of lipid droplets within cells. In addition, cells were collected to assess the expression of adipogenic markers and RARγ.
10
Lentiviral Transduction of miR-135b in MCF-7 Cells
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Lentiviral particles, expressing a control miRNA or hsa-miR-135b double strand precursors, were obtained from Genechem (Shanghai, China). MCF-7 cell lines, stably expressing control miRNA or hsa-miR-135b, were established by transducing cells with Lentiviral particles, followed by selection with puromycin.
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