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Pmirtarget vector

Manufactured by Promega
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The PMirTarget vector is a plasmid designed for the expression and evaluation of miRNA (microRNA) targets. It provides a platform for studying the interactions between miRNAs and their target sequences.

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Lab products found in correlation

Pmirtarget vector, by OriGene (2 mentions) Prl renilla luciferase control reporter vector, by Promega (1 mentions) Dual luciferase reporter assay, by Promega (1 mentions) N luminometer, by Turner Designs (1 mentions) Dual luciferase assay system, by Promega (1 mentions)

2 protocols using pmirtarget vector

1

miR-21-5p Regulation of SMAD7 3'UTR

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The complete sequence of SMAD7 3′ untranslated region (UTR) was constructed into the pMirTarget vector (Origene, Rockville, MD, USA). The plasmid containing the putative binding sequence of miR-21-5p “ATAAGCTA” is named as “SMAD7 3′UTR”. For luciferase assay, 50 nm of control miRNA or miR-21-5p mimics, 0.8 µg of pMirTarget vector containing SMAD7 3′ UTR, and 0.08 µg of pRL renilla luciferase control reporter vector (Promega, Madison, WI, USA) were co-transfected into HEK-293 cells in 96-well plates for 48 h. Cell were harvested and analyzed using the dual-luciferase reporter assay (Promega) according to the manufacturer’s instruction.
Chang C.H., Yen M.C., Liao S.H., Hsu Y.L., Lai C.S., Kuo Y.R, & Hsu Y.L. (2017). Dual Role of MiR-21-Mediated Signaling in HUVECs and Rat Surgical Flap under Normoxia and Hypoxia Condition. International Journal of Molecular Sciences, 18(9), 1917.
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2

Luciferase Assay for MET and EGFR 3'UTR

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The 2,300 bp 3’-UTR of the human c-MET or 1,700 bp 3’-UTR of the human EGFR was cloned into the pMirTarget vector (Origene, Rockville, MD) to produce the 3-UTR-luciferase plasmids. The 3’-UTR-c-MET, 3’UTR-EGFR or the empty pMirTarget vector was co-transfected with miR-206 mimic (100 nM) in SKOV3 and A498 cells, and luciferase activities were measured in cell lysates using the Dual Luciferase Assay System (Promega Corporation, Madison, WI) with a 20/20n luminometer (Turner Designs, Sunnyvale, CA) [57 ].
Choi B.H., Ryu D.Y., Ryoo I.G, & Kwak M.K. (2017). NFE2L2/NRF2 silencing-inducible miR-206 targets c-MET/EGFR and suppresses BCRP/ABCG2 in cancer cells. Oncotarget, 8(63), 107188-107205.
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