Efficient Transgene‐Free Multiplexed Germline Editing via Viral Delivery of an Engineered <scp>TnpB</scp>

T.W., M.K, H.S., J.A.D. and S.E.J. have filed patents related to this work. S.E.J. is a cofounder and consultant for Inari Agriculture and a consultant for Terrana Biosciences, Invaio Sciences, Sail Biomedicines and Zymo Research. J.A.D. is a cofounder of Azalea Therapeutics, Caribou Biosciences, Editas Medicine, Evercrisp, Scribe Therapeutics and Mammoth Biosciences, a scientific advisory board member at Evercrisp, Caribou Biosciences, Scribe Therapeutics, Mammoth Biosciences, The Column Group and Inari, an advisor for Aditum Bio, the Chief Science Advisor to Sixth Street, a Director at Johnson & Johnson, Altos and Tempus, and has a research project sponsored by Apple Tree Partners. Amp-seq data is accessible at NCBI Sequence Read Archive BioProject PRJNA1427739. RNA-seq data is available at GEO accession: GSE316183. Figure S1: Co-delivery of RNA1 and two RNA2 vectors does not enable multiplexed genome editing. Figure S2: Development of Ymu1 multiplexed gRNA arrays for plant gene editing. Figure S3: TRV-delivery of Ymu1 for multiplexed genome editing. Table S1: Genotype characterisation of green progeny from plants infected with TRV expressing Ymu1 and gRNAs targeting AtCHLl1 gRNA4 and AtPDS3 gRNA12 (pTW2278_54 and pTW2278_17). Table S2: Plasmids and their descriptions used in this study. Table S3: Target sites and primers used in this study. Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.