Generation of an hiPSC-1 knock-in line expressing TY1-tagged MNX1-protein together with mScarlet

Abstract

MNX1 encodes a homeobox transcription factor with conserved embryonic requirements in spinal motor neuron formation and pancreatic beta-cell differentiation. Mutations in MNX1 are associated with dominantly inherited Currarino syndrome and neonatal diabetes. To better understand embryonic MNX1 functions we generated an hiPSC-1 knock-in line heterozygously expressing MNX1 C-terminally tagged with 2xTY1 together with a T2A-separated red fluorescent reporter mScarlet. The TY1 epitope tag was introduced to enable immunoprecipitation based analyses on molecular MNX1 interactions and mScarlet was included for enrichment of MNX1 expressing cell populations. This cell line shows normal karyotype, pluripotency marker expression and differentiation potential in vitro.

Keywords

• Karyotyping report
• Mycoplasma PCR-test
• Sequencing results
• Southern blots
• Plasmid sequences of gRNA-Cas9 vectors and targeting construct