Generation and Correction of iPSCs with MAPT Mutation

Dermal fibroblasts from MAPT p.R406W carriers (F11362 and F11421) were transduced with non-integrating Sendai virus carrying OCT3/4, SOX2, KLF4, and cMYC (Life Technologies) as previously described [19 (link)]. iPSCs that were heterozygous for MAPT p.R406W were edited to WT (F11362.1Δ1B06 and F11421.12Δ2A07) using CRISPR/Cas9 as previously reported [19 (link), 39 ]. Mutation status was confirmed by Sanger sequencing (Supplemental Fig. 1). Cell lines were maintained in mTeSR medium (StemCell Technologies) on Matrigel. Cell lines were confirmed to be free of mycoplasma.

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Mahali S., Martinez R., King M., Verbeck A., Harari O., Benitez B.A., Horie K., Sato C., Temple S, & Karch C.M. (2022). Defective proteostasis in induced pluripotent stem cell models of frontotemporal lobar degeneration. Translational Psychiatry, 12, 508.

Publication 2022

OCT3/4 mTeSR medium

Cell lines Crispr Fibroblasts Heterozygous Ipscs Klf4 Mapt Matrigel Mutation Mycoplasma Oct3 Sendai virus Sox2 Stemcell

Corresponding Organization :

Other organizations : Washington University in St. Louis, Neural Stem Cell Institute, Hope Center for Neurological Disorders

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Variable analysis

independent variables

Transduction of dermal fibroblasts from MAPT p.R406W carriers with non-integrating Sendai virus carrying OCT3/4, SOX2, KLF4, and cMYC
CRISPR/Cas9 editing of iPSCs heterozygous for MAPT p.R406W to wild-type (WT)

dependent variables

Generation of induced pluripotent stem cells (iPSCs) from dermal fibroblasts

control variables

Cell lines were maintained in mTeSR medium on Matrigel
Cell lines were confirmed to be free of mycoplasma

controls

Positive control: Dermal fibroblasts from MAPT p.R406W carriers (F11362 and F11421)
Negative control: Not explicitly mentioned

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