Generate iNeurons

Stable insertion of dox-inducible NGN1,2 cassette in CLYBL safe-harbor locus in pluripotent stem cell line

Custom iNeuron lines will be generated from a client-specific iPSC line. This involves electroporation of TALEN plasmids (pZT-C13-R1 and pZT-C13-L1) targeting CLYBL and a homology-directed repair cassette (see map below). Individual mCherry positive iPSCs are cloned and 2-3 resulting clonal lines will be selected for 1) Good iPSC morphology, 2) iNeuron induction, 3) correct insertion assessed by PCR. The project takes 6-8 weeks.