An Inducible Expression System of the Calcium-Activated Potassium
Channel 4 to Study the Differential Impact on Embryonic Stem Cells
Figure 2
Construction of the iSK4 cell line. (a) Scheme of the expression cassette integrated into the desired loci. The ES cell line A2lox.cre harbors an rtTA in the constitutive ROSA26 locus on chromosome 6. A neomycin (noeR) resistance is recombined upon integration of the targeting vector. Recombination of the construct into the prepared site on the X-chromosome is facilitated by Cre. Transgenic cells can subsequently be selected by G418 (neo). tetOP: tetracycline response element; PGK: phosphoglycero kinase promoter; ATG: methionine initiation codon; black triangle: lox recognition sequence for Cre recombinase; GFP: green fluorescent protein; Neo: truncated neomycin (G418) resistance gene; pA: polyadenylation sequence. (b) Lox-in PCR verifies the correct integration of the expression cassette at determined loci on chr6 by amplifying a product at the size of ~450 bp. Untargeted ES cells were used as a control.
