Pre-mademicroRNAexrepssionlentivirusforhumanhsa-miR-125b-5p(MIMAT0000423):
The humanmicroRNA precursorsanditsnativecontextsequences(upstreamanddownstreamflankinggenomicsequences)wasclonedintoaGentarget’smiRNAexpressionlentivector: pLenti-TetCMV(GFP-3UTR/miRNA)-Rsv(Puro).ThemiRNAclonedat3’UTRorGFPMarkerunderanoptionalinducIBLeCMVpromoter. ItcanbeusedforconstitutivemiRexpressionorusedasinduciblemiRexpressionwhentheTetRrepressorisprescent.TheGFPsignalprovidesareal-timemonitorforthemiRNAexpressionlevelsandthelentivirustransductionefficiency.
TheclonedmiRNAexpressprecursorsequenceislistedbelow.Note: theRed-highlightedregionisthemiRStem-loopsequence:
ACCATACCACCTGTTTGTTGCATCTTAGTTATGAACCTCGAACAGAAATTGCCTGTCATT
CTTGTTTTGCTTTGCTTTGTCTCAAGAAAGAAAACATTGTTGCGCTCCTCTCAGTCCCTG
AGACCCTAACTTGTGATGTTTACCGTTTAAATCCACGGGTTAGGCTCTTGGGAGCTGCG
AGTCGTGCTTTTGCATCCTGGAAATTTGGTGGAATTTTATTCTTTAAAGCAAAAACAAAA
GAAAAGAAAGTTTGTCTGAGGTGATTGAGTATACCTCTGAGGTTTTCAT
ItguaranteestogeneratethematuredmiRNAsequenceasfollows:
UCCCUGAGACCCUAACUUGUGA
See Producemanual fordetails.
Amounts:
1)500ulof ready-to-usespecificmiRNAexpressionlentivirusinDMEM-mediumcontaining60ugpolybrene(10x).
2)500ulof ready-to-use Negative-controlmiRNAexpressionlentivirusinDMEM-mediumcontaining60ugpolybrene(10x).(Note:ThemiR-Control doesnottranscribeanymiRNAsequences,buthasthesamebackbonewithGFPmarker).
Virustiter: 1×107 IFU/ml,
Shipping: overnightshipmentindry-icepackage.(Note:Thisproductispre-madeinstock,readytoshipviaovernightdry-icepackage)