PT  - JOURNAL ARTICLE
AU  - James E. Voss
AU  - Alicia Gonzalez-Martin
AU  - Raiees Andrabi
AU  - Roberta P. Fuller
AU  - Ben Murrell
AU  - Laura E. McCoy
AU  - Katelyn Porter
AU  - Deli Huang
AU  - Wenjuan Li
AU  - Devin Sok
AU  - Khoa Le
AU  - Bryan Briney
AU  - Morgan Chateau
AU  - Geoffrey Rogers
AU  - Lars Hangartner
AU  - Ann J. Feeney
AU  - David Nemazee
AU  - Paula Cannon
AU  - Dennis R. Burton
TI  - Reprogramming the antigen specificity of B cells using genomeediting technologies
AID  - 10.1101/455402
DP  - 2018 Jan 01
TA  - bioRxiv
PG  - 455402
4099  - http://biorxiv.org/content/early/2018/10/29/455402.short
4100  - http://biorxiv.org/content/early/2018/10/29/455402.full
AB  - We have developed a method to introduce novel paratopes into the human antibody repertoire by modifying the immunoglobulin genes of mature B cells directly using genome editing technologies. We used CRISPR-Cas9 in a homology directed repair strategy, to replace the heavy chain (HC) variable region in B cell lines with that from an HIV broadly neutralizing antibody, PG9. Our strategy is designed to function in cells that have undergone VDJ recombination using any combination of variable (V), diversity (D) and joining (J) genes. The modified locus expresses PG9 HC which pairs with native light chains resulting in the cell surface expression of HIV specific B cell receptors (BCRs). Endogenous activation-induced cytidine deaminase (AID) in engineered cells allowed for Ig class switching and generated BCR variants with improved anti-HIV neutralizing activity. Thus, BCRs engineered in this way retain the genetic flexibility normally required for affinity maturation during adaptive immune responses.