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

NIH RePORTER · NIH · R21 · $263,250 · view on reporter.nih.gov ↗

Abstract

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.

Key facts

NIH application ID
10947198
Project number
1R21OD037648-01
Recipient
OREGON HEALTH & SCIENCE UNIVERSITY
Principal Investigator
Benjamin Bimber
Activity code
R21
Funding institute
NIH
Fiscal year
2024
Award amount
$263,250
Award type
1
Project period
2024-07-01 → 2026-06-30