Products/Services Used | Details | Operation |
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Plasmid DNA Preparation> | All-in-one Cas9/sgRNA plasmid pSpCas9 BB-2A-Puro encoding sgRNA specific for CycT1 (sgRNA sequence: AAGC AGATTGGCCGCCTGC) (GenScript) was transfected into 293T cells using Lipofectamine 2000 (Invitrogen). | Get A Quote |
Transcription of HIV provirus is a key step of the viral cycle, and depends on the recruitment of the cellular positive transcription elongation factor b (P-TEFb) to the HIV promoter. The viral transactivator Tat can displace P-TEFb from the 7SK small nuclear ribonucleoprotein, where it is bound and inactivated by HEXIM1, and bring it to TAR, which allows the stalled RNA polymerase II to transition to successful transcription elongation. In this study, we designed a chimeric inhibitor of HIV transcription by combining functional domains from HEXIM1 and Tat. The chimera (HT1) potently inhibited gene expression from the HIV promoter, by competing with Tat for TAR and P-TEFb binding, while keeping the latter inact... More