GFP-on mouse model for interrogation of in vivo gene editing

Abstract Gene editing technologies have revolutionized therapies for numerous genetic diseases. However, in vivo gene editing hinges on identifying efficient delivery vehicles for editing in targeted cell types, a significant hurdle in fully realizing its therapeutic potential. A model system to rap...

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Main Authors: Carla Dib, Jack A. Queenan, Leah Swartzrock, Hana Willner, Morgane Denis, Nouraiz Ahmed, Fareha Moulana Zada, Beltran Borges, Carsten T. Charlesworth, Tony Lum, Bradley P. Yates, Caleb Y. Kwon, Augustino V. Scorzo, Scott C. Davis, Jessie R. Davis, Ran He, Jun Xie, Guangping Gao, Tippi C. MacKenzie, David R. Liu, Gregory A. Newby, Agnieszka D. Czechowicz
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Nature Communications
Online Access:https://doi.org/10.1038/s41467-025-61449-y
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author Carla Dib
Jack A. Queenan
Leah Swartzrock
Hana Willner
Morgane Denis
Nouraiz Ahmed
Fareha Moulana Zada
Beltran Borges
Carsten T. Charlesworth
Tony Lum
Bradley P. Yates
Caleb Y. Kwon
Augustino V. Scorzo
Scott C. Davis
Jessie R. Davis
Ran He
Jun Xie
Guangping Gao
Tippi C. MacKenzie
David R. Liu
Gregory A. Newby
Agnieszka D. Czechowicz
author_facet Carla Dib
Jack A. Queenan
Leah Swartzrock
Hana Willner
Morgane Denis
Nouraiz Ahmed
Fareha Moulana Zada
Beltran Borges
Carsten T. Charlesworth
Tony Lum
Bradley P. Yates
Caleb Y. Kwon
Augustino V. Scorzo
Scott C. Davis
Jessie R. Davis
Ran He
Jun Xie
Guangping Gao
Tippi C. MacKenzie
David R. Liu
Gregory A. Newby
Agnieszka D. Czechowicz
author_sort Carla Dib
collection DOAJ
description Abstract Gene editing technologies have revolutionized therapies for numerous genetic diseases. However, in vivo gene editing hinges on identifying efficient delivery vehicles for editing in targeted cell types, a significant hurdle in fully realizing its therapeutic potential. A model system to rapidly evaluate systemic gene editing would advance the field. Here, we develop the GFP-on reporter mouse, which harbors a nonsense mutation in a genomic EGFP sequence correctable by adenine base editor (ABE) among other genome editors. The GFP-on system was validated using single and dual adeno-associated virus (AAV9) encoding ABE8e and sgRNA. Intravenous administration of AAV9-ABE8e-sgRNA into adult GFP-on mice results in EGFP expression consistent with the tropism of AAV9. Intrahepatic delivery of AAV9-ABE8e-sgRNA into GFP-on fetal mice restores EGFP expression in AAV9-targeted organs lasting at least six months post-treatment. The GFP-on model provides an ideal platform for high-throughput evaluation of emerging gene editing tools and delivery modalities.
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spelling doaj-art-0f78f85ddd1848a486dbab685e88a4242025-08-20T03:46:15ZengNature PortfolioNature Communications2041-17232025-07-0116111310.1038/s41467-025-61449-yGFP-on mouse model for interrogation of in vivo gene editingCarla Dib0Jack A. Queenan1Leah Swartzrock2Hana Willner3Morgane Denis4Nouraiz Ahmed5Fareha Moulana Zada6Beltran Borges7Carsten T. Charlesworth8Tony Lum9Bradley P. Yates10Caleb Y. Kwon11Augustino V. Scorzo12Scott C. Davis13Jessie R. Davis14Ran He15Jun Xie16Guangping Gao17Tippi C. MacKenzie18David R. Liu19Gregory A. Newby20Agnieszka D. Czechowicz21Department of Pediatrics, Division of Hematology, Oncology, Stem Cell Transplantation and Regenerative Medicine, Stanford University School of MedicineMerkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and HarvardDepartment of Pediatrics, Division of Hematology, Oncology, Stem Cell Transplantation and Regenerative Medicine, Stanford University School of MedicineDepartment of Pediatrics, Division of Hematology, Oncology, Stem Cell Transplantation and Regenerative Medicine, Stanford University School of MedicineDepartment of Pediatrics, Division of Hematology, Oncology, Stem Cell Transplantation and Regenerative Medicine, Stanford University School of MedicineMerkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and HarvardDepartment of Surgery, University of California San FranciscoDepartment of Surgery, University of California San FranciscoInstitute for Stem Cell Biology and Regenerative Medicine, Stanford UniversityDepartment of Surgery, University of California San FranciscoDepartment of Genetic Medicine, Johns Hopkins University School of MedicineDartmouth College, Thayer School of EngineeringDartmouth College, Thayer School of EngineeringDartmouth College, Thayer School of EngineeringMerkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and HarvardDepartment of Genetics & Cellular Medicine, University of Massachusetts Chan Medical SchoolDepartment of Genetics & Cellular Medicine, University of Massachusetts Chan Medical SchoolDepartment of Genetics & Cellular Medicine, University of Massachusetts Chan Medical SchoolDepartment of Surgery, University of California San FranciscoMerkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and HarvardMerkin Institute of Transformative Technologies in Healthcare, Broad Institute of MIT and HarvardDepartment of Pediatrics, Division of Hematology, Oncology, Stem Cell Transplantation and Regenerative Medicine, Stanford University School of MedicineAbstract Gene editing technologies have revolutionized therapies for numerous genetic diseases. However, in vivo gene editing hinges on identifying efficient delivery vehicles for editing in targeted cell types, a significant hurdle in fully realizing its therapeutic potential. A model system to rapidly evaluate systemic gene editing would advance the field. Here, we develop the GFP-on reporter mouse, which harbors a nonsense mutation in a genomic EGFP sequence correctable by adenine base editor (ABE) among other genome editors. The GFP-on system was validated using single and dual adeno-associated virus (AAV9) encoding ABE8e and sgRNA. Intravenous administration of AAV9-ABE8e-sgRNA into adult GFP-on mice results in EGFP expression consistent with the tropism of AAV9. Intrahepatic delivery of AAV9-ABE8e-sgRNA into GFP-on fetal mice restores EGFP expression in AAV9-targeted organs lasting at least six months post-treatment. The GFP-on model provides an ideal platform for high-throughput evaluation of emerging gene editing tools and delivery modalities.https://doi.org/10.1038/s41467-025-61449-y
spellingShingle Carla Dib
Jack A. Queenan
Leah Swartzrock
Hana Willner
Morgane Denis
Nouraiz Ahmed
Fareha Moulana Zada
Beltran Borges
Carsten T. Charlesworth
Tony Lum
Bradley P. Yates
Caleb Y. Kwon
Augustino V. Scorzo
Scott C. Davis
Jessie R. Davis
Ran He
Jun Xie
Guangping Gao
Tippi C. MacKenzie
David R. Liu
Gregory A. Newby
Agnieszka D. Czechowicz
GFP-on mouse model for interrogation of in vivo gene editing
Nature Communications
title GFP-on mouse model for interrogation of in vivo gene editing
title_full GFP-on mouse model for interrogation of in vivo gene editing
title_fullStr GFP-on mouse model for interrogation of in vivo gene editing
title_full_unstemmed GFP-on mouse model for interrogation of in vivo gene editing
title_short GFP-on mouse model for interrogation of in vivo gene editing
title_sort gfp on mouse model for interrogation of in vivo gene editing
url https://doi.org/10.1038/s41467-025-61449-y
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