Figure 1. Representation of RNAi subcloning strategy. Modified pSUPER (MunI) vector carrying the H1-RNAi cassette: a H1-RNA promoter driving a shRNA after BglII-HindIII cloning of the encoding dsDNA oligo. From this EcoRI-MunI digested vector, the H1-RNAi cassette is cloned into EcoRI digested pTRIPΔU3-CMV-EGFP-WPRE transfer plasmid, leaving one EcoRI site upstream for cloning of additional H1-RNAi cassettes. Figure shows relevant part of transfer, packaging and envelope plasmids, which are co-transfected for virus production. CMV: Cytomegalovirus minimal promoter; LTR: long terminal repeat; ΔLTR: partial deleted LTR; RRE: Rev responsive element; cPPT: central polypurine tract; WPRE: Woodchuck Hepatitis Virus Posttranscriptional Regulatory Element; VSV-G: Vesicular Stomatitis Virus Glycoprotein; polyA: polyadenylation signal; bp: base pairs.