ENAapplication/xmlftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR315/046/SRR31559646/SRR31559646_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR305/010/SRR30578110/SRR30578110_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR305/012/SRR30578112/SRR30578112_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR305/011/SRR30578111/SRR30578111_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR305/010/SRR30578110/SRR30578110_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR305/014/SRR30578114/SRR30578114_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR305/013/SRR30578113/SRR30578113_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR305/011/SRR30578111/SRR30578111_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR315/045/SRR31559645/SRR31559645_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR315/047/SRR31559647/SRR31559647_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR315/045/SRR31559645/SRR31559645_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR305/015/SRR30578115/SRR30578115_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR305/014/SRR30578114/SRR30578114_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR315/047/SRR31559647/SRR31559647_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR315/046/SRR31559646/SRR31559646_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR305/012/SRR30578112/SRR30578112_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR305/013/SRR30578113/SRR30578113_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR305/015/SRR30578115/SRR30578115_1.fastq.gzprimaryOK200GenomicsTao Lab, Regenerative Medicine and Cell Biology, Medical University of South Carolinahttps://www.ebi.ac.uk/ena/browser/view/PRJNA1157886Homo sapiensxref:PubMed:39768143Over 200 point mutations in ryanodine receptor (RyR2) of cardiac sarcoplasmic reticulum (SR) are known to associate with cardiac arrhythmia. We have already reported on the calcium signaling phenotype of a point mutation in RyR2 Ca2+ binding site Q3925E expressed in human stem-cell derived cardiomyocytes (hiPSC-CMs) that was found to be lethal in a 9 year old girl. CRISPR/Cas9 gene edited mutant cardiomyocytes carrying RyR2-Q3925E mutation exhibited loss of calcium-induced calcium release (CICR) and caffeine-triggered calcium release, but continued to beat arrhythmically without generating significant SR Ca2+ release, consistent with remodeling of calcium signaling pathway. RNAseq heat-map confirmed significant changes of calcium associated genes, supporting the remodeling possibility. To determine the in-situ cardiac phenotype in animal model of this mutation, we generated a knock-in mouse model of RyR2-Q3924E+/– using CRISPR/Cas9 technique. We obtained 3 homozygous and one chimera mice, but they all died before 3 weeks of age preventing the establishment of germline mutation transmission in the offsprings. Histo-pathological analysis of the heart showed significant cardiac hypertrophy suggesting Q3924E-RyR2 mutation was lethal in the mouse. Overall design: We sought to utilize hiPSC-CMs to recapitulate the previously identified Q3925E and E3848A mutations using CRISPR/Cas9 gene editingENAfalseAttempts to create transgenic mice carrying the Q3925E mutation in RyR2 Ca2+ binding siteAttempts to create transgenic mice carrying the Q3925E mutation in RyR2 Ca2+ binding site2025-09-242025-01-03PRJNA1157886GSE276596960639768143