Rapid Generation of Transgenic Rhesus Macaques using a Safe-harbor Genomic Docking Site

SUMMARY
It is now possible to generate nonhuman primates (NHPs) with precise germline genomic alterations, enabling
researchers to generate highly refined models of human disease. Significant investments by our team at the
Oregon National Primate Research Center (ONPRC) have greatly advanced NHP transgenic technology,
resulting in the births of five healthy, genetically altered rhesus macaques (RM) since 2019. While these
advancements are extremely promising, existing methods to create transgenic NHPs are too slow and
expensive for most research applications. Innovative technologies are needed to accelerate creation of
transgenic NHPs.
Here we propose to adapt site-directed recombination, a proven technology used in rodent transgenics, to
fundamentally advance NHP transgenic generation. We will generate founder animals with a small “docking
site” integrated into the AAVS1 safe-harbor locus, which is a well-characterized genomic site that provides
consistent expression across tissues. Gametes collected and banked from these animals will provide an “off-
the shelf" system for subsequent gene editing and transgenic model creation. Embryos or cells from animals
encoding this acceptor site can be injected/transfected with a donor vector that contains the desired genetic
cargo. Site-directed recombination mediates integration of the cargo into AAVS1, with high on-target efficiency
and few limitations in cargo size. This technology has the potential to revolutionize the non-human primate
transgenic field.

Grant Number: 5R21OD037648-02
NIH Institute/Center: NIH
Principal Investigator: Benjamin Bimber