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. 2023 Feb 16;29(2):412–421. doi: 10.1038/s41591-022-02190-7

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© The Author(s) 2023

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Extended Data Fig. 1 — A) Schematics of AAV9 genomes containing fluorescent reporter genes (not to scale) used to assess dual transduction. ITR: AAV9 inverted terminal repeats. Tnnt2 prom: cardiomyocyte specific troponin T promoter. CMV prom: Cytomegalovirus promoter, a strong, constitutive promoter. WPRE: Woodchuck hepatitis virus post-transcriptional regulatory element, used to stabilize transcripts. pA-Term: polyadenylation signal and transcriptional terminator. B) Fluorescent micrographs of ventricular section (short axis view) and liver from a WT mouse injected with 3×10¹³ vg/kg AAV2/9-Tnnt2-eGFP.RBG (Tnnt2-eGFP.RBG) and AAV2/9-CMV-mScarlet.WPRE.bGH (CMV-mScarlet.WPRE.bGH) and imaged with a fluorescence microscope. Scale bar: 1 mm. Imaging revealed co-expression of green fluorescent protein (top panel) delivered by AAV9-Tnnt2-eGFP.RBG and red fluorescent protein (middle panel) delivered by AAV9-CMV-mScarlet.WPRE.bGH are co-localized (yellow signal, lower panel) in cardiac cells. Representative images are shown from two technical replicates.