Engineering Protein-Secreting Plasma Cells by Homology-Directed Repair in Primary Human B Cells.

ABSTRACT: The ability to engineer primary human B cells to differentiate into long-lived plasma cells and secrete a de novo protein may allow the creation of novel plasma cell therapies for protein deficiency diseases and other clinical applications. We initially developed methods for efficient genome editing of primary B cells isolated from peripheral blood. By delivering CRISPR/CRISPR-associated protein 9 (Cas9) ribonucleoprotein (RNP) complexes under conditions of rapid B cell expansion, we achieved site-specific gene disruption at multiple loci in primary human B cells (with editing rates of up to 94%). We used this method to alter ex vivo plasma cell differentiation by disrupting developmental regulatory genes. Next, we co-delivered RNPs with either a single-stranded DNA oligonucleotide or aden

SUBMITTER: Hung KL 

PROVIDER: S-EPMC5835153 | biostudies-literature | 2018 Feb

REPOSITORIES: biostudies-literature