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. 2014 May 19;9(5):e97764. doi: 10.1371/journal.pone.0097764

Figure 1. Overview of the GATEWAY-compatible lentiviral tetracycline-regulated RNAi (GLTR) RNAi system A).

Figure 1

The H1-RNA core promoter constitutively drives the expression of a downstream shRNA encoding gene. TetO sequences (circles) allow the tetra−/doxycycline-dependent binding of TetR or TetR fusion proteins such as TetR-KRAB. B) GATEWAY compatible ENTR-plasmids for constitutive (pENTR-H1) and conditional (pENTR-THT and -THT-III) expression of shRNAs. C) GATEWAY compatible lentiviral destination (DEST) vectors. The fluorescence protein (FP) vectors encode GFP or RFP, the selectable vectors (S) puromycin or neomycin resistance. The one-vector system pGLTR-X encodes sequences for the simultaneous expression of GFP or puromycin resistance and the TetR-repressor. Abbreviations: H1, RNA-polymerase III promoter; THT, TetO sequences flanked H1 promoter; att (L and R) attachment site; KanR, kanamycin resistance; GentaR, gentamycin resistance; LTR, long terminal repeat; RRE, rev response element; cPPT, central polypurine tract; CmR, chloramphenicol resistance; ccdB, ccdB protein (inhibitor of E.coli DNA gyrase); SFFV U3, spleen focus forming virus U3 promoter; WPRE, woodchuck hepatitis posttranscriptional response element; AmpR, ampicillin resistance; T2A, 2A-like cis-acting hydrolase elements of Thosea asigna virus; P2A, 2A-like cis-acting hydrolase elements of porcine teschovirus.