Generation of an induced pluripotent stem cell line (TRNDi002-B) from a patient carrying compound heterozygous p.Q208X and p.G310G mutations in the NGLY1 gene

Abstract

NGLY1 deficiency is a rare genetic disease caused by mutations in the NGLY1 gene that encodes N-glycanase 1. The disease phenotype in patient cells is unclear. A human induced pluripotent stem cell (iPSC) line was generated from skin dermal fibroblasts of a patient with NGLY1 deficiency that has compound heterozygous mutations of a p.Q208X variant (c.622C > T) in exon 4 and a p.G310G variant (c.930C > T) in exon 6 of the NGLY1 gene. This iPSC line offers a useful resource to study the disease pathophysiology and a cell-based model for drug development to treat NGLY1 deficiency.

An Induced Pluripotent Stem Cell‐Derived Neuromuscular Junction Platform for Study of the NGLY1‐Congenital Disorder of Deglycosylation

“…iPSCs were generated previously from the dermal fibroblasts of healthy and NGLY1 deficient patients [ 19 ] as well as a line of iPSCs that had been CRISPR modified to possess a homozygous knockout of the NGLY1 gene. In the context of this paper, the lines will be referred to as 268A (WT), X2‐9 (Isogenic KO), 592D (Patient 1), and 594A (Patient 2).…”

Generation of an induced pluripotent stem cell line (TRNDi008-A) from a Hunter syndrome patient carrying a hemizygous 208insC mutation in the IDS gene

“…As described previously (Li et al, 2019), patient fibroblasts were reprogrammed into iPS cells using non-integrating Sendai virus vector technology (A16517, ThermoFisher).…”

“…Total RNA was extracted using RNeasy Plus Mini Kit (Qiagen) and 1 μg of RNA was reverse transcribed into cDNA with Superscript ™ III First-Strand Synthesis SuperMix kit. PCR was performed using Platinum II Hot-Start PCR Master Mix (ThermoFisher) and the amplifications were carried out as previously described (Li et al, 2019) using the primers listed in Table 2. Products were then loaded to the E -Gel ® 1.2% with SYBR Safe ™ gel and imaged using a G: Box Chemi-XX6 gel doc system (Syngene).…”

An induced pluripotent stem cell line (TRNDi006-A) from a MPS IIIB patient carrying homozygous mutation of p.Glu153Lys in the NAGLU gene

“…Genomic analysis of NAGLU variants was performed by Applied StemCell (Milpitas, California). Genomic DNA was extracted from TRNDi006-A using QuickExtract™ DNA Extraction Solution (Lucigen) and PCR amplifications using MyTaq™ Red Mix (Bioline) were carried out using a previously defined protocol (Li et al, 2018). Sanger sequencing analysis was used for genotyping of the homozygous mutation for a p. Glu153Lys variant (c.457 G > A) of the NAGLU gene.…”