Human iPSCs used in this study have been previously described [20 ]. iPSC lines were generated using non-integrating Sendai virus carrying the Yamanaka factors: OCT3/4, SOX2, KLF4, and cMYC (Life Technologies) [21 , 22 (link)]. The following parameters were used for the characterization of each of the iPSC lines using standard methods [21 ]: pluripotency markers by immunocytochemistry (ICC) and quantitative PCR (qPCR); spontaneous or TriDiff differentiation into the three germ layers by ICC and qPCR; assessment of chromosomal abnormalities by karyotyping; and MAPT mutation status confirmation by Sanger sequencing (characterization data previously reported [15 (link)]).
To determine the impact of the MAPT mutant allele on molecular phenotypes, we used CRISPR/Cas9-edited isogenic controls in which the mutant allele was reverted to the wild-type (WT) allele in each of the donor iPSC lines as previously described [15 (link), 20 ]. The resulting edited iPSC lines were characterized as described above in addition to on- and off-target sequencing (characterization data previously reported [15 (link)]). All iPSC lines used in this study carry the MAPT H1/H1 common haplotype. All cell lines were confirmed to be free of mycoplasma.
To determine the impact of the MAPT mutant allele on molecular phenotypes, we used CRISPR/Cas9-edited isogenic controls in which the mutant allele was reverted to the wild-type (WT) allele in each of the donor iPSC lines as previously described [15 (link), 20 ]. The resulting edited iPSC lines were characterized as described above in addition to on- and off-target sequencing (characterization data previously reported [15 (link)]). All iPSC lines used in this study carry the MAPT H1/H1 common haplotype. All cell lines were confirmed to be free of mycoplasma.
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