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. 2013 Oct 11;8(10):e76279. doi: 10.1371/journal.pone.0076279

Figure 2. Overview of pLEG/pREG vectors to express shRNAmirs.

Figure 2

A) A typical four-plasmid LR recombination reaction showing the insertion of a gene (i), selection marker (ii) and miRNA cassette (iii) into pLEG(R1–R4) (iv) to produce a recombinant lentiviral virus (v). B) Schematic of the miRNA cassette and entry plasmid showing the Chloramphenicol resistance/ccdB cell death cassette situated between XhoI/EcoRI sites of pBEG miRNA(R3-ccdB-L4) to increase the cloning efficiency of novel shRNAs. C) The retroviral destination vector pREG(R1–R4) used in four-plasmid LR recombination reactions – functions as in (A). KanR: Kanamycin resistance gene; 5′MIR: 5′miR30 sequences; Cmr: chloramphenicol resistance marker; 3′MIR: 3′miR30 sequences.