Rat HCs were isolated from isogeneic rats, as described [33 (link)]. The isolated HCs were transduced with lentivirus carrying either null (as a control) or recombinant HIF-1a under a CMV promoter (Origene, Shanghai, China). For HC reinfusion, 107 donor null/HIF-1a-transduced HCs were given to the receipt rats via tail vein 3 days before IR.
Cmv promoter
Manufactured by OriGene
Sourced in China
The CMV promoter is a DNA sequence that functions as a strong promoter, driving high-level expression of downstream genes in a wide variety of cell types. It is commonly used in molecular biology and genetic engineering applications to achieve efficient transcription of target genes.
Automatically generated - may contain errors
Lab products found in correlation
Lentivirus, by OriGene (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
C57 b6j mice, by Jackson ImmunoResearch (1 mentions)
T4 dna ligase, by New England Biolabs (1 mentions)
Dual luciferase assay, by Promega (1 mentions)
Pmirtarget luciferase vector, by OriGene (1 mentions)
Biotek synergy 2, by Agilent Technologies (1 mentions)
Lipofectamine, by Thermo Fisher Scientific (1 mentions)
6 protocols using cmv promoter
1
Rat Hepatocyte Transduction and Reinfusion
2
NGLY1 Overexpression in Cells
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A pLenti expression vector that carries a Myc-DDK-tagged-human NGLY1 open reading frame driven by a CMV promoter (OriGene Technologies, Rockville, MD) was transduced into cells for the overexpression of NGLY1. A pLenti-C-Myc-DDK empty vector was used as the transduction control.
3
Hematopoietic Cell Transduction for Wound Healing
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Mouse hematopoietic cells were isolated from isogeneic mice, as described [29 (link)]. Mouse blood cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 20% fetal bovine serum (Invitrogen, Carlsbad, CA, USA) in a humidified chamber with 5% CO2 at 37 °C. The isolated blood cells were transduced with lentivirus carrying either null (as a control) or recombinant HIF-1a under a CMV promoter (Origene, Shanghai, China). For blood cell self-transfusion, 107 donor null/HIF-1a-transduced blood cells were re-infused into the circulation of receipt mice via tail vein at the same time of induction of skin wound.
4
Untagged ADSL Expression Vector
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An expression vector in which reading frame of ADSL is placed under the control of CMV promoter was obtained from Origene. In this expression vector ADSL is presented as un-tagged protein in order to avoid artefacts arising due to N or C terminal tags. Restriction enzymes with respective buffers and T4DNA ligase were from NEB. Trans-IT invivo gene delivery solution was from mirus biosciences. C57B6J mice were from Jackson laboratories USA.
5
Plasmid Construction and Purification
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The plasmid pUbC-GFP expresses GFP from the ubiquitin C (UbC) promoter (Addgene, Cambridge, MA). pCMV-Cav1 expresses a GFP- tagged mouse caveolae protein 1 (Cav1) from the CMV promoter (Origene, Rockville, MD), and pUbC-Cav1 expresses a GFP-tagged mouse Cav1 from the ubiquitin (UbC) promoter cloned from pUbC-GFP and pCMV-Cav1 plasmids. All plasmids were purified using Qiagen Giga-prep kits (Qiagen, Chatsworth, CA) and suspended in 10 mM Tris-HCl (pH 8.0), 1 mM ethylenediaminetetraacetic acid, and 140 mM NaCl.
6
Luciferase Assay for miR-10a Regulation
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The overall length 3'-UTR of PI3K catalytic subunit α(PIK3CA) was subcloned into a pMir-Target luciferase vector that promoted by a cytomegalovirus (CMV) promoter (Origene, Rockville, MD, USA). Mutant derivatives of the structure were fabricated by site-directed mutagenesis by the Quick-Change Kit (Stratagene, La Jolla, CA, USA). HEK293 cells were cotransfected with the luciferase structure (wild type or mutant) with miR-10a mimics or control scrambled siRNA for 48 h using Lipofectamine (Invitrogen) at a 2:1 molar ratio (miRNA mimics vs. construct reporter) according to the manufacturer's instruction. The cells were counted and reseeded into 96-well assay plates 24h before luciferase dection, followed by dual-luciferase assay according to the instruction (Promega, Madison, WI, USA). Luminescence was detected by a multimode microplate reader (BioTek Synergy 2; BioTek, Winooski, VT, USA).
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