SHOX2 c.*28T>C hiPSC sib-selection #1, gene-edited using gRNA-1
Homo sapiens
We  generated hiPSC derived from peripheral blood mononuclear cells of two AF  patients harboring heterozygous SHOX2 mutations (SHOX2 c.849C>A and SHOX2  c.*28T>C) using non-integrating Sendai viruses coding KLF4, OCT3/4, SOX2,  c-MYC. All derived clones displayed stem cell-like features and pluripotency  capacity, had lost the viral transgenes after 10-20 passages, expressed  pluripotency markers on protein and RNA level and differentiated  spontaneously into all three germ layers. Moreover, they were karyotypically  normal and had a confirmed patient origin. Isogenic pairs showed equal  differentiation capacity into three major cardiac subtypes (pacemaker, atrial  and ventricular), no gross sarcomeric aberration and equal subtype-specific  marker gene expression. To correct the heterozygous SHOX2 mutations, cells  transfected with Cas9 protein, gRNAs and single-stranded DNA (repair  template) were seeded in small pools of 200 cells/well ('sib-selection') and  deep sequenced. An above-average amount of WT alleles indicated the presence  of isogenic cells in this sib-selection which could be isolated by  single-cell sequencing.
isolate
Index patient II harboring a heterozygous SHOX2 c.*28T>C mutation
age
41
biomaterial_provider
Alessandra Moretti, First Department of Medicine, Cardiology, Klinikum rechts der Isar Technical University of Munich, 81675 Munich, Bavaria, Germany
sex
male
tissue
blood sample
cell_line
SHOX2 c.*28T>C sib-selection 1-1
cell_type
induced pluripotent stem cells
disease
atrial fibrillation
disease_stage
persistent
ethnicity
white
health_state
diseased
karyotype
46,XY
phenotype
full pluripotency capacity, normal karyotype
race
caucasian
sample_type
single cell-derived clone
strain
hiPSC
genotype
SHOX2 c.*28T>C (-/-)
geo_loc_name
Germany
growth_protocol
StemFit medium, according to manufacturer's recommendation
store_cond
extraction from cultured cells, stored a -20C
BioSampleModel
Human