描述

pLVX-shRNA1 is an HIV-1-based, lentiviral expression vector designed to express a small hair-pin RNA (shRNA) for RNA interference (RNAi) studies. Expression of your shRNA is drivenby the RNA Pol III-dependent, human U6 promoter (PU6), located just upstream of the MCS. pLVX-shRNA1 can either be used as a plasmid expression vector and transfected into cells, or it can be packaged into viral particles and transduced into cells. Lentiviral particles derived from the vector allow the expression of shRNAs in virtually any cell type, including primary cells. pLVX-shRNA1 contains all of the viral processing elements necessary for the production of replication-incompetent lentivirus, as well as elements to improve viral titer and overall vector function. The woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) promotes RNA processing events and enhances nuclear export of viral RNA (1), leading to increased viral titers from packaging cells. In addition, the vector includes a Rev-response element (RRE), which further increases viral titers by enhancing the transport of unspliced viral RNA out of the nucleus (2). Finally, pLVX-shRNA1 also contains a central polypurine tract/central termination sequence element (cPPT/CTS). During target cell infection, thiselement creates a central DNA flap that increases nuclear import of the viral genome, resulting in improved vector integration and more efficient transduction (3). In addition to lentiviral elements, pLVX-shRNA1 contains a puromycin resistance gene (Puror)under the control of the murine phosphoglycerate kinase promoter (PPGK) for the selection of stable transductants. The vector also contains a pUC origin of replication and an E. coli ampicillin resistance gene (Ampr) for propagation and selection in bacteria.

應(yīng)用

pLVX-shRNA1 is available in the Lenti-X? shRNA Expression System (Cat. No. 632177). Before it can be transduced into target cells, the vector must be cotransfected into 293T cells with our Lenti-X HTX Packaging System (Cat. Nos. 631247 and 631249) and packaged into viral particles. This packaging system allows the safe production of high titer, infectious, replication-incompetent, VSV-G pseudotyped lentiviral particles that can infect a wide range of cell types, including non-dividing and primary cells (4).

0001 TGGAAGGGCT AATTCACTCC CAAAGAAGAC AAGATATCCT TGATCTGTGG ATCTACCACA 
0061 CACAAGGCTA CTTCCCTGAT TAGCAGAACT ACACACCAGG GCCAGGGGTC AGATATCCAC 
0121 TGACCTTTGG ATGGTGCTAC AAGCTAGTAC CAGTTGAGCC AGATAAGGTA GAAGAGGCCA 
0181 ATAAAGGAGA GAACACCAGC TTGTTACACC CTGTGAGCCT GCATGGGATG GATGACCCGG 
0241 AGAGAGAAGT GTTAGAGTGG AGGTTTGACA GCCGCCTAGC ATTTCATCAC GTGGCCCGAG 
0301 AGCTGCATCC GGAGTACTTC AAGAACTGCT GATATCGAGC TTGCTACAAG GGACTTTCCG 
0361 CTGGGGACTT TCCAGGGAGG CGTGGCCTGG GCGGGACTGG GGAGTGGCGA GCCCTCAGAT 
0421 CCTGCATATA AGCAGCTGCT TTTTGCCTGT ACTGGGTCTC TCTGGTTAGA CCAGATCTGA 
0481 GCCTGGGAGC TCTCTGGCTA ACTAGGGAAC CCACTGCTTA AGCCTCAATA AAGCTTGCCT 
0541 TGAGTGCTTC AAGTAGTGTG TGCCCGTCTG TTGTGTGACT CTGGTAACTA GAGATCCCTC 
0601 AGACCCTTTT AGTCAGTGTG GAAAATCTCT AGCAGTGGCG CCCGAACAGG GACTTGAAAG 
0661 CGAAAGGGAA ACCAGAGGAG CTCTCTCGAC GCAGGACTCG GCTTGCTGAA GCGCGCACGG 
0721 CAAGAGGCGA GGGGCGGCGA CTGGTGAGTA CGCCAAAAAT TTTGACTAGC GGAGGCTAGA 
0781 AGGAGAGAGA TGGGTGCGAG AGCGTCAGTA TTAAGCGGGG GAGAATTAGA TCGCGATGGG 
0841 AAAAAATTCG GTTAAGGCCA GGGGGAAAGA AAAAATATAA ATTAAAACAT ATAGTATGGG 
0901 CAAGCAGGGA GCTAGAACGA TTCGCAGTTA ATCCTGGCCT GTTAGAAACA TCAGAAGGCT 
0961 GTAGACAAAT ACTGGGACAG CTACAACCAT CCCTTCAGAC AGGATCAGAA GAACTTAGAT 
1021 CATTATATAA TACAGTAGCA ACCCTCTATT GTGTGCATCA AAGGATAGAG ATAAAAGACA 
1081 CCAAGGAAGC TTTAGACAAG ATAGAGGAAG AGCAAAACAA AAGTAAGACC ACCGCACAGC 
1141 AAGCGGCCGG CCGCTGATCT TCAGACCTGG AGGAGGAGAT ATGAGGGACA ATTGGAGAAG 
1201 TGAATTATAT AAATATAAAG TAGTAAAAAT TGAACCATTA GGAGTAGCAC CCACCAAGGC 
1261 AAAGAGAAGA GTGGTGCAGA GAGAAAAAAG AGCAGTGGGA ATAGGAGCTT TGTTCCTTGG 
1321 GTTCTTGGGA GCAGCAGGAA GCACTATGGG CGCAGCGTCA ATGACGCTGA CGGTACAGGC 
1381 CAGACAATTA TTGTCTGGTA TAGTGCAGCA GCAGAACAAT TTGCTGAGGG CTATTGAGGC 
1441 GCAACAGCAT CTGTTGCAAC TCACAGTCTG GGGCATCAAG CAGCTCCAGG CAAGAATCCT 
1501 GGCTGTGGAA AGATACCTAA AGGATCAACA GCTCCTGGGG ATTTGGGGTT GCTCTGGAAA 
1561 ACTCATTTGC ACCACTGCTG TGCCTTGGAA TGCTAGTTGG AGTAATAAAT CTCTGGAACA 
1621 GATTTGGAAT CACACGACCT GGATGGAGTG GGACAGAGAA ATTAACAATT ACACAAGCTT 
1681 AATACACTCC TTAATTGAAG AATCGCAAAA CCAGCAAGAA AAGAATGAAC AAGAATTATT 
1741 GGAATTAGAT AAATGGGCAA GTTTGTGGAA TTGGTTTAAC ATAACAAATT GGCTGTGGTA 
1801 TATAAAATTA TTCATAATGA TAGTAGGAGG CTTGGTAGGT TTAAGAATAG TTTTTGCTGT 
1861 ACTTTCTATA GTGAATAGAG TTAGGCAGGG ATATTCACCA TTATCGTTTC AGACCCACCT 
1921 CCCAACCCCG AGGGGACCCG ACAGGCCCGA AGGAATAGAA GAAGAAGGTG GAGAGAGAGA 
1981 CAGAGACAGA TCCATTCGAT TAGTGAACGG ATCTCGACGG TATCGCCTTT AAAAGAAAAG 
2041 GGGGGATTGG GGGGTACAGT GCAGGGGAAA GAATAGTAGA CATAATAGCA ACAGACATAC 
2101 AAACTAAAGA ATTACAAAAA CAAATTACAA AAATTCAAAA TTTTCGGGTT TATTACAGGG 
2161 ACAGCAGAGA TCCAGTTTAT CGATCTGGGC AGGAAGAGGG CCTATTTCCC ATGATTCCTT 
2221 CATATTTGCA TATACGATAC AAGGCTGTTA GAGAGATAAT TAGAATTAAT TTGACTGTAA 
2281 ACACAAAGAT ATTAGTACAA AATACGTGAC GTAGAAAGTA ATAATTTCTT GGGTAGTTTG 
2341 CAGTTTTAAA ATTATGTTTT AAAATGGACT ATCATATGCT TACCGTAACT TGAAAGTATT 
2401 TCGATTTCTT GGCTTTATAT ATCTTGTGGA AAGGACGAGG ATCCGGACAA GCTTCGAATT 
2461 CTACCGGGTA GGGGAGGCGC TTTTCCCAAG GCAGTCTGGA GCATGCGCTT TAGCAGCCCC 
2521 GCTGGGCACT TGGCGCTACA CAAGTGGCCT CTGGCCTCGC ACACATTCCA CATCCACCGG 
2581 TAGGCGCCAA CCGGCTCCGT TCTTTGGTGG CCCCTTCGCG CCACCTTCTA CTCCTCCCCT 
2641 AGTCAGGAAG TTCCCCCCCG CCCCGCAGCT CGCGTCGTGC AGGACGTGAC AAATGGAAGT 
2701 AGCACGTCTC ACTAGTCTCG TGCAGATGGA CAGCACCGCT GAGCAATGGA AGCGGGTAGG 
2761 CCTTTGGGGC AGCGGCCAAT AGCAGCTTTG CTCCTTCGCT TTCTGGGCTC AGAGGCTGGG 
2821 AAGGGGTGGG TCCGGGGGCG GGCTCAGGGG CGGGCTCAGG GGCGGGGCGG GCGCCCGAAG 
2881 GTCCTCCGGA GGCCCGGCAT TCTGCACGCT TCAAAAGCGC ACGTCTGCCG CGCTGTTCTC 
2941 CTCTTCCTCA TCTCCGGGCC TTTCGACCTG CAGCCCAAGC TTACCATGAC CGAGTACAAG 
3001 CCCACGGTGC GCCTCGCCAC CCGCGACGAC GTCCCCAGGG CCGTACGCAC CCTCGCCGCC 
3061 GCGTTCGCCG ACTACCCCGC CACGCGCCAC ACCGTCGATC CGGACCGCCA CATCGAGCGG 
3121 GTCACCGAGC TGCAAGAACT CTTCCTCACG CGCGTCGGGC TCGACATCGG CAAGGTGTGG 
3181 GTCGCGGACG ACGGCGCCGC GGTGGCGGTC TGGACCACGC CGGAGAGCGT CGAAGCGGGG 
3241 GCGGTGTTCG CCGAGATCGG CCCGCGCATG GCCGAGTTGA GCGGTTCCCG GCTGGCCGCG 
3301 CAGCAACAGA TGGAAGGCCT CCTGGCGCCG CACCGGCCCA AGGAGCCCGC GTGGTTCCTG 
3361 GCCACCGTCG GCGTCTCGCC CGACCACCAG GGCAAGGGTC TGGGCAGCGC CGTCGTGCTC 
3421 CCCGGAGTGG AGGCGGCCGA GCGCGCCGGG GTGCCCGCCT TCCTGGAGAC CTCCGCGCCC 
3481 CGCAACCTCC CCTTCTACGA GCGGCTCGGC TTCACCGTCA CCGCCGACGT CGAGGTGCCC 
3541 GAAGGACCGC GCACCTGGTG CATGACCCGC AAGCCCGGTG CCTGACGCCC GCCCCACGAC 
3601 CCGCAGCGCC CGACCGAAAG GAGCGCACGA CCCCATGCAT CGGCGTCTCG AGTCTAGAGG 
3661 GCCCGACGCG TCTGGAACAA TCAACCTCTG GATTACAAAA TTTGTGAAAG ATTGACTGGT 
3721 ATTCTTAACT ATGTTGCTCC TTTTACGCTA TGTGGATACG CTGCTTTAAT GCCTTTGTAT 
3781 CATGCTATTG CTTCCCGTAT GGCTTTCATT TTCTCCTCCT TGTATAAATC CTGGTTGCTG 
3841 TCTCTTTATG AGGAGTTGTG GCCCGTTGTC AGGCAACGTG GCGTGGTGTG CACTGTGTTT 
3901 GCTGACGCAA CCCCCACTGG TTGGGGCATT GCCACCACCT GTCAGCTCCT TTCCGGGACT 
3961 TTCGCTTTCC CCCTCCCTAT TGCCACGGCG GAACTCATCG CCGCCTGCCT TGCCCGCTGC 
4021 TGGACAGGGG CTCGGCTGTT GGGCACTGAC AATTCCGTGG TGTTGTCGGG GAAGCTGACG 
4081 TCCTTTCCAT GGCTGCTCGC CTGTGTTGCC ACCTGGATTC TGCGCGGGAC GTCCTTCTGC 
4141 TACGTCCCTT CGGCCCTCAA TCCAGCGGAC CTTCCTTCCC GCGGCCTGCT GCCGGCTCTG 
4201 CGGCCTCTTC CGCGTCTTCG CCTTCGCCCT CAGACGAGTC GGATCTCCCT TTGGGCCGCC 
4261 TCCCCGCCTG GAATTAATTC TGCAGTCGAG ACCTAGAAAA ACATGGAGCA ATCACAAGTA 
4321 GCAATACAGC AGCTACCAAT GCTGATTGTG CCTGGCTAGA AGCACAAGAG GAGGAGGAGG 
4381 TGGGTTTTCC AGTCACACCT CAGGTACCTT TAAGACCAAT GACTTACAAG GCAGCTGTAG 
4441 ATCTTAGCCA CTTTTTAAAA GAAAAGAGGG GACTGGAAGG GCTAATTCAC TCCCAACGAA 
4501 GACAAGATAT CCTTGATCTG TGGATCTACC ACACACAAGG CTACTTCCCT GATTAGCAGA 
4561 ACTACACACC AGGGCCAGGG GTCAGATATC CACTGACCTT TGGATGGTGC TACAAGCTAG 
4621 TACCAGTTGA GCCAGATAAG GTAGAAGAGG CCAATAAAGG AGAGAACACC AGCTTGTTAC 
4681 ACCCTGTGAG CCTGCATGGG ATGGATGACC CGGAGAGAGA AGTGTTAGAG TGGAGGTTTG 
4741 ACAGCCGCCT AGCATTTCAT CACGTGGCCC GAGAGCTGCA TCCGGAGTAC TTCAAGAACT 
4801 GCTGATATCG AGCTTGCTAC AAGGGACTTT CCGCTGGGGA CTTTCCAGGG AGGCGTGGCC 
4861 TGGGCGGGAC TGGGGAGTGG CGAGCCCTCA GATCCTGCAT ATAAGCAGCT GCTTTTTGCC 
4921 TGTACTGGGT CTCTCTGGTT AGACCAGATC TGAGCCTGGG AGCTCTCTGG CTAACTAGGG 
4981 AACCCACTGC TTAAGCCTCA ATAAAGCTTG CCTTGAGTGC TTCAAGTAGT GTGTGCCCGT 
5041 CTGTTGTGTG ACTCTGGTAA CTAGAGATCC CTCAGACCCT TTTAGTCAGT GTGGAAAATC 
5101 TCTAGCAGTA GTAGTTCATG TCATCTTATT ATTCAGTATT TATAACTTGC AAAGAAATGA 
5161 ATATCAGAGA GTGAGAGGCC TTGACATTGC TAGCGTTTAC CGTCGACCTC TAGCTAGAGC 
5221 TTGGCGTAAT CATGGTCATA GCTGTTTCCT GTGTGAAATT GTTATCCGCT CACAATTCCA 
5281 CACAACATAC GAGCCGGAAG CATAAAGTGT AAAGCCTGGG GTGCCTAATG AGTGAGCTAA 
5341 CTCACATTAA TTGCGTTGCG CTCACTGCCC GCTTTCCAGT CGGGAAACCT GTCGTGCCAG 
5401 CTGCATTAAT GAATCGGCCA ACGCGCGGGG AGAGGCGGTT TGCGTATTGG GCGCTCTTCC 
5461 GCTTCCTCGC TCACTGACTC GCTGCGCTCG GTCGTTCGGC TGCGGCGAGC GGTATCAGCT 
5521 CACTCAAAGG CGGTAATACG GTTATCCACA GAATCAGGGG ATAACGCAGG AAAGAACATG 
5581 TGAGCAAAAG GCCAGCAAAA GGCCAGGAAC CGTAAAAAGG CCGCGTTGCT GGCGTTTTTC 
5641 CATAGGCTCC GCCCCCCTGA CGAGCATCAC AAAAATCGAC GCTCAAGTCA GAGGTGGCGA 
5701 AACCCGACAG GACTATAAAG ATACCAGGCG TTTCCCCCTG GAAGCTCCCT CGTGCGCTCT 
5761 CCTGTTCCGA CCCTGCCGCT TACCGGATAC CTGTCCGCCT TTCTCCCTTC GGGAAGCGTG 
5821 GCGCTTTCTC ATAGCTCACG CTGTAGGTAT CTCAGTTCGG TGTAGGTCGT TCGCTCCAAG 
5881 CTGGGCTGTG TGCACGAACC CCCCGTTCAG CCCGACCGCT GCGCCTTATC CGGTAACTAT 
5941 CGTCTTGAGT CCAACCCGGT AAGACACGAC TTATCGCCAC TGGCAGCAGC CACTGGTAAC 
6001 AGGATTAGCA GAGCGAGGTA TGTAGGCGGT GCTACAGAGT TCTTGAAGTG GTGGCCTAAC 
6061 TACGGCTACA CTAGAAGAAC AGTATTTGGT ATCTGCGCTC TGCTGAAGCC AGTTACCTTC 
6121 GGAAAAAGAG TTGGTAGCTC TTGATCCGGC AAACAAACCA CCGCTGGTAG CGGTGGTTTT 
6181 TTTGTTTGCA AGCAGCAGAT TACGCGCAGA AAAAAAGGAT CTCAAGAAGA TCCTTTGATC 
6241 TTTTCTACGG GGTCTGACGC TCAGTGGAAC GAAAACTCAC GTTAAGGGAT TTTGGTCATG 
6301 AGATTATCAA AAAGGATCTT CACCTAGATC CTTTTAAATT AAAAATGAAG TTTTAAATCA 
6361 ATCTAAAGTA TATATGAGTA AACTTGGTCT GACAGTTACC AATGCTTAAT CAGTGAGGCA 
6421 CCTATCTCAG CGATCTGTCT ATTTCGTTCA TCCATAGTTG CCTGACTCCC CGTCGTGTAG 
6481 ATAACTACGA TACGGGAGGG CTTACCATCT GGCCCCAGTG CTGCAATGAT ACCGCGAGAC 
6541 CCACGCTCAC CGGCTCCAGA TTTATCAGCA ATAAACCAGC CAGCCGGAAG GGCCGAGCGC 
6601 AGAAGTGGTC CTGCAACTTT ATCCGCCTCC ATCCAGTCTA TTAATTGTTG CCGGGAAGCT 
6661 AGAGTAAGTA GTTCGCCAGT TAATAGTTTG CGCAACGTTG TTGCCATTGC TACAGGCATC 
6721 GTGGTGTCAC GCTCGTCGTT TGGTATGGCT TCATTCAGCT CCGGTTCCCA ACGATCAAGG 
6781 CGAGTTACAT GATCCCCCAT GTTGTGCAAA AAAGCGGTTA GCTCCTTCGG TCCTCCGATC 
6841 GTTGTCAGAA GTAAGTTGGC CGCAGTGTTA TCACTCATGG TTATGGCAGC ACTGCATAAT 
6901 TCTCTTACTG TCATGCCATC CGTAAGATGC TTTTCTGTGA CTGGTGAGTA CTCAACCAAG 
6961 TCATTCTGAG AATAGTGTAT GCGGCGACCG AGTTGCTCTT GCCCGGCGTC AATACGGGAT 
7021 AATACCGCGC CACATAGCAG AACTTTAAAA GTGCTCATCA TTGGAAAACG TTCTTCGGGG 
7081 CGAAAACTCT CAAGGATCTT ACCGCTGTTG AGATCCAGTT CGATGTAACC CACTCGTGCA 
7141 CCCAACTGAT CTTCAGCATC TTTTACTTTC ACCAGCGTTT CTGGGTGAGC AAAAACAGGA 
7201 AGGCAAAATG CCGCAAAAAA GGGAATAAGG GCGACACGGA AATGTTGAAT ACTCATACTC 
7261 TTCCTTTTTC AATATTATTG AAGCATTTAT CAGGGTTATT GTCTCATGAG CGGATACATA 
7321 TTTGAATGTA TTTAGAAAAA TAAACAAATA GGGGTTCCGC GCACATTTCC CCGAAAAGTG 
7381 CCACCTGACG TCGACGGATC GGGAGATCAA CTTGTTTATT GCAGCTTATA ATGGTTACAA 
7441 ATAAAGCAAT AGCATCACAA ATTTCACAAA TAAAGCATTT TTTTCACTGC ATTCTAGTTG 
7501 TGGTTTGTCC AAACTCATCA ATGTATCTTA TCATGTCTGG ATCAACTGGA TAACTCAAGC 
7561 TAACCAAAAT CATCCCAAAC TTCCCACCCC ATACCCTATT ACCACTGCCA ATTACCTGTG 
7621 GTTTCATTTA CTCTAAACCT GTGATTCCTC TGAATTATTT TCATTTTAAA GAAATTGTAT 
7681 TTGTTAAATA TGTACTACAA ACTTAGTAGT TTTTAAAGAA ATTGTATTTG TTAAATATGT 
7741 ACTACAAACT TAGTAGT