The development of the AI-generated gene-editing enzyme OpenCRISPR-1 (OC-1) circumvents long-standing intellectual property disputes surrounding classic CRISPR-Cas9, which have previously hindered the clinical application of cell-based therapies. In the effort to generate hypoimmune cells, we demonstrate the high-efficiency knockout of B2M and CIITA genes using OC-1 in both plasmid and protein formats across multiple cell types, including induced pluripotent stem cells (iPSCs), primary mesenchymal stem cells (MSCs), and MSCs derived from iPSCs (iMSCs). These results were validated via TIDE (Tracking of Indels by Decomposition) analysis and flow cytometry, confirming the generation of stable knockout clones. The resulting hypoimmune iPSCs were confirmed by sequencing and flow cytometry. Furthermore, we expanded the utility of OC-1 by establishing ‘Landing Pad' cell lines using a site-specific integration system (Bxb1 integrase) at genomic safe harbor loci. Additionally, the use of a single-stranded DNA (ssODN) donor template enables a streamlined, one-step insertion of genes of interest (GOI) into specific genomic sites with high precision.
About the speaker:
Dr. Yongming (Luke) Ren has over a decade of experience in the stem cell field, spanning both academic and industrial settings. His work includes several influential publications and a patent application focused on the manufacture of therapeutic cells for muscular dystrophy. Most recently, at REPROCELL, he led the investigation into applying OpenCRISPR technology within stem cell research.