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. 2006 Apr 11;94(9):1300–1310. doi: 10.1038/sj.bjc.6603094

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Copyright © 2006 Cancer Research UK

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Scheme of pSUPER.retro vector. A specific sequence for anti-COX-2 short hairpin-RNA (sense sequence – loop – antisense sequence) was cloned into pSUPER.retro vector and the transcription of this sequence is regulated by H1 promoter for RNA pol-III. GFP gene expression provides a rapid test for infection efficiency and the gene for puromycin resistance is necessary to select clones expressing shRNAs against COX-2 mRNA. All cassettes are included into retroviral 5′–3′ LTRs to allow provirus integration in host cells genome. 3′ LTR is inactivated by deletion to avoid virus replication inside infected cells (see details under Materials And Methods).