Multiplexed-CREATE Selection Yields AAV Vectors Targeting Different Cell Types of the Central Nervous System Following Systemic Delivery

Abstract

Recombinant adeno-associated viral (rAAV) capsids are widely accepted as safe gene delivery vehicles in research laboratories and in gene therapy clinical trials and there is potential to further improve their usage by evolving the surface of the capsids to enhance their affinity to specific cell-types or tissues after intravenous (IV) delivery. To this end, we built upon our previous Cre recombination-based AAV targeted evolution (CREATE) method (Deverman et al, Nat. Biotech., 2016; Chan et al., Nat. Neurosci., 2017) to develop Multiplexed-CREATE (M-CREATE). M-CREATE facilitates both positive and negative selections and minimizes the propagation of biases from successive rounds of selection via synthetic library generation. In addition to increasing the confidence in the selections, M-CREATE enables a detailed characterization of the selection process, improving our understanding of selection progression and outcome.

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