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GenParts™ DNA Fragments> | … The resulting HDR-edited cells could be enriched with a reporter gene and exhibited … method for gene knockin in human Tregs to generate a uniform population of gene-edited cells. As … DNA, plasmids, or synthesized (IDT gBlocks or GenScript GenParts), then cloned by Gibson … | Get A Quote |
Regulatory T cell (Treg) therapy is a promising curative approach for a variety of immune-mediated conditions. CRISPR-based genome editing allows precise insertion of transgenes through homology-directed repair, but its use in human Tregs has been limited. We report an optimized protocol for CRISPR-mediated gene knockin in human Tregs with high-yield expansion. To establish a benchmark of human Treg dysfunction, we target the master transcription factor FOXP3 in naive and memory Tregs. Although FOXP3-ablated Tregs upregulate cytokine expression, effects on suppressive capacity in vitro manifest slowly and primarily in memory Tregs. Moreover, FOXP3-ablated Tregs retain their characteristic protein, transcripti... More