Pre-mademicroRNAexrepssionlentivirusforhumanhsa-miR-34a-5p(MIMAT0000255):
The humanmicroRNA precursorsanditsnativecontextsequences(upstreamanddownstreamflankinggenomicsequences)wasclonedintoaGentarget’smiRNAexpressionlentivector: pLenti-TetCMV(GFP-3UTR/miRNA)-Rsv(Puro).ThemiRNAclonedat3’UTRorGFPMarkerunderanoptionalinducIBLeCMVpromoter. ItcanbeusedforconstitutivemiRexpressionorusedasinduciblemiRexpressionwhentheTetRrepressorisprescent.TheGFPsignalprovidesareal-timemonitorforthemiRNAexpressionlevelsandthelentivirustransductionefficiency.
TheclonedmiRNAexpressprecursorsequenceislistedbelow.Note: theRed-highlightedregionisthemiRStem-loopsequence:
TGCATCCTTTCTTTCCTCCCCACATTTCCTTCTTATCAACAGGTGCTGGGGAGAGGCAGGACAGGCCTGTCCCCCGA
GTCCCCTCCGGATGCCGTGGACCGGCCAGCTGTGAGTGTTTCTTTGGCAGTGTCTTAGCTGGTTGTTGTGAGCAAT
AGTAAGGAAGCAATCAGCAAGTATACTGCCCTAGAAGTGCTGCACGTTGTGGGGCCCAAGAGGGAAGATGAAG
CGAGAGATGCCCAGACCAGTGGGAGACGCCAGGACTTCGGAAGCTCTTCTGCGCCACGGTGGGTGGTGAGGGC
GGCTGGGAAAG
ItguaranteestogeneratetwomaturedmiRNAsequencesasfollows:
UGGCAGUGUCUUAGCUGGUUGU (miR#112)
CAAUCAGCAAGUAUACUGCCCU (miR#113)
See Producemanual fordetails.
Amounts:
1)500ulof ready-to-usespecificmiRNAexpressionlentivirusinDMEM-mediumcontaining60ugpolybrene(10x).
2)500ulof ready-to-use Negative-controlmiRNAexpressionlentivirusinDMEM-mediumcontaining60ugpolybrene(10x).(Note:ThemiR-ControlexpresstheGFPmarker,butnotcontainanymiRNAsequence).
Virustiter: 1×107 IFU/ml,
Shipping: overnightshipmentindry-icepackage.(Note:Thisproductispre-madeinstock,readytoshipviaovernightdry-icepackage)