Speaker 1: Khaled Alsayegh
Title: Large deletions during gene editing in human pluripotent stem cells

Abstract:
Human pluripotent stem cells (hPSCs) hold great promise for being a versatile source of cells for regenerative medicine. When coupled with CRISPR/Cas9 gene-editing technologies, the potential of hPSCs in cell therapy and disease modeling could be augmented. To harness their full potential, detailed understanding of the DNA repair mechanisms during gene editing in these cells is essential. In this context, we analyze DNA repair outcomes in hPSCs following CRISPR/Cas9-induced double strand breaks using PacBio sequencing.

Speaker 2: Chongwei Bi
Title: Single-molecule Sequencing for Detecting Rare Genetic Variants in Human Pluripotent Stem Cells after CRISPR Genome Editing

Abstract:
Genome contains hereditary information of the species, and its integrity is vital to life. With the developing of next-generation sequencing (NGS) technologies, we are gaining an unprecedented advance in decoding the genomic information in recent years. However, the current sequencing strategies are mainly population-based, in which samples come from DNA pools of bulk cells. They are not suitable for detecting rare mutations in a subpopulation of cells. Here we introduce targeted individual DNA molecule sequencing (IDMseq), which is a strategy to label and sequence individual DNA molecules from a pool. We applied IDMseq to detect rare mutations. Results showed that IDMseq is broadly applicable to any high-throughput sequencing platforms, and is capable to detect ultra-rare variants, including small variants and large structural variants, at 1:10,000 allele frequency.