TIGER: A tdTomato in vivo genome- editing reporter mouse for investigating precision- editor delivery approaches

• 成果类型: Article
• 署名作者: Du, Samuel W.; Palczewska, Grazyna; Dong, Zhiqian; Lauterborn, Julie C.; Kaipa, Balasankara Reddy; Yan, Alexander L.; Holubowicz, Rafa; Ha, Siyoung; Chen, Paul Z.; Gall, Christine M.; Zode, Gulab; Liu, David R.; Palczewski, Krzysztof
• 署名单位: University of California System; University of California Irvine; University of California System; University of California Irvine; University of California System; University of California Irvine; Harvard University; Massachusetts Institute of Technology (MIT); Broad Institute; Harvard University; Howard Hughes Medical Institute; Harvard University; Massachusetts Institute of Technology (MIT); University of California System; University of California Irvine; University of California System; University of California Irvine
• 刊物名称: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• ISSN/ISSBN: 0027-14496
• DOI: 10.1073/pnas.2506257122
• 发表日期: 2025-09-02
• 关键词: fluorescent protein; gene; base; mutations; red; dna

摘要

In vivo genome editing has the potential to address many inherited and environmental disorders. However, a major hurdle for the clinical translation of genome editing is safe, efficient delivery to disease-relevant tissues. A modality-agnostic reporter animal model that facilitates rapid, precise, and quantifiable assessment of functional delivery and editing could greatly enhance the evaluation and translation of delivery technologies. Here, we present the development of the tdTomato in vivo genome-editing reporter (TIGER) mouse, a reporter strain that harbors an integrated and constitutively expressed mutated tdTomato gene in the Polr2a locus. The mutations (Q115X, Q357X) abolish fluorescence, but successful adenine base editing (ABE) or prime editing (PE) restores tdTomato fluorescence. This mouse model facilitates the tissue-and cell type-specific assessment of genome editing agent delivery. We describe several editing strategies validated in vitro and demonstrate efficient ABE and PE in vivo using viral and nonviral delivery vectors targeting four cell types within the mouse eye: the retinal pigment epithelium, photoreceptors, M & uuml;ller glia, and the trabecular meshwork. We show direct editing characterization in the ocular tissues via in vivo and ex vivo two-photon confocal microscopy and verify the spectral and fluorescence lifetime properties of tdTomato reporter in other mouse tissues. Additionally, we demonstrate successful adeno-associated virus (AAV)-mediated PE of extraocular tissues, including hepatocytes, skeletal muscle, and brain neurons by intravenous injection. Thus, the TIGER mouse facilitates the direct development, comparison, and optimization of delivery platforms for efficient and productive ABE or PE broadly applicable in vivo across multiple tissues tested in this study.