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. Author manuscript; available in PMC: 2015 Jan 1.
Published in final edited form as: Methods Mol Biol. 2014;1181:229–247. doi: 10.1007/978-1-4939-1047-2_20

Fig. 1.

Fig. 1

Zinc finger nuclease (ZFN)-mediated targeting of the GCaMP3 expression cassette to the AAVS1 locus. (a) The AAVS1 ZFN plasmid includes two expression cassettes, in which separate human PGK promoters drive the expression of the AAVS1 site-specific ZFN pair. (b) The pZDonor-GCaMP3 plasmid includes GCaMP3 and neomycin selection cassettes that are flanked by AAVS1 site homology arms (each ~800 bp). GCaMP3 expression is driven by the CAG promoter, while the neomycin resistance gene is driven by the human PGK promoter. (c ) The AAVS locus is located between exons 1 and 2 of the PPR1R12C gene on chromosome 19. (d) Co-transfection of AAVS1 ZFN and ZDonor-GCaMP3 plasmids in human cells results in targeting of the latter vector to the AAVS1 locus by homologous recombination (see Subheading 3.1 for details). pA bovine growth hormone polyadenylation signal, Ex exon, NeoKan neomycin/kanamycin resistance