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Transposase-assisted target-site integration for efficient plant genome engineering

2024/06/26 by Peng Liu, Kaushik Panda, Seth A. Edwards +12 · 1 voice · 96 citations
Agricultural and Biological Sciences · Biochemistry, Genetics and Molecular Biology · #Biology #CRISPR #CRISPR and Genetic Engineering #Cas9 #Chromosomal and Genetic Variations #Computational biology #Enhancer #Gene #Genetics #Genome #Genome editing #Genome engineering #Insert (composites) #Plant Virus Research Studies #Transposable element #Transposase

paper · pdf · doi:10.1038/s41586-024-07613-8

published in Nature 631(8021), 593-600 (Nature Portfolio)

openalex publication_date 2024/06/26 · openalex created_date 2025/10/10 · openalex updated_date 2026/08/06

Abstract

Abstract The current technologies to place new DNA into specific locations in plant genomes are low frequency and error-prone, and this inefficiency hampers genome-editing approaches to develop improved crops 1,2 . Often considered to be genome ‘parasites’, transposable elements (TEs) evolved to insert their DNA seamlessly into genomes 3–5 . Eukaryotic TEs select their site of insertion based on preferences for chromatin contexts, which differ for each TE type 6–9 . Here we developed a genome engineering tool that controls the TE insertion site and cargo delivered, taking advantage of the natural ability of the TE to precisely excise and insert into the genome. Inspired by CRISPR-associated transposases that target transposition in a programmable manner in bacteria 10–12 , we fused the rice Pong transposase protein to the Cas9 or Cas12a programmable nucleases. We demonstrated sequence-specific targeted insertion (guided by the CRISPR gRNA) of enhancer elements, an open reading frame and a gene expression cassette into the genome of the model plant Arabidopsis . We then translated this system into soybean—a major global crop in need of targeted insertion technology. We have engineered a TE ‘parasite’ into a usable and accessible toolkit that enables the sequence-specific targeting of custom DNA into plant genomes.

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