Skip to main content
. 2010 Jun 28;107(28):12475–12480. doi: 10.1073/pnas.1001402107

Fig. 2.

Fig. 2.

Schematic of library selection and clonal analysis. (i) Packaging of library viruses via transfection of plasmid library and VSV-G helper plasmid into 293T cells. (ii) Harvest and purification of viruses. (iii) Infection of 293T cells with virion library at a multiplicity of infection (moi) of < 0.1. (iv) Sorting and subsequent expansion of GFP-positive cells. (v) Rescue of packageable mutants by 293T cell transfection with helper plasmid encoding the VSV-G envelope protein. (vi) Subcloning Gag-Pol.ZF cDNA from packageable mutants into pCMV Gag-Pol helper plasmid. (vii) Packaging virus clones via transfection of 293T cells with pCLPIT GFP and helper plasmids. (viii) Analysis of infectivity and genome-wide integration analysis.