RRC ID 81817
Author Furuhata Y, Kimura M, Sakai A, Murakami T, Egi E, Sakuma T, Yamamoto T, Yoshizumi T, Kato Y.
Title Direct protein delivery into intact Arabidopsis cells for genome engineering.
Journal Sci Rep
Abstract Intracellular delivery of biomolecules is a prerequisite for genetic techniques such as recombinant engineering and genome editing. Realizing the full potential of this technology requires the development of safe and effective methods for delivering protein tools into cells. In this study, we demonstrated the spontaneous internalization of exogenous proteins into intact cells and root tissue of whole plants of Arabidopsis thaliana. We termed this internalization phenomenon as protein Delivery Independent of Vehicles or Equipment (DIVE), which efficiently delivered genome engineering proteins including Cre recombinase and zinc-finger nucleases (ZFN) into plant cells. Using protein DIVE, we achieved less toxic protein delivery than electroporation with up to 94% efficiency in Arabidopsis cell culture and 19% genome modification in Arabidopsis plants that was maintained in regenerated tissue. This work illustrates the potential of protein DIVE for a wide range of applications, including genome engineering in plants.
Volume 14(1)
Pages 22568
Published 2024-9-29
DOI 10.1038/s41598-024-72978-9
PII 10.1038/s41598-024-72978-9
PMID 39343787
PMC PMC11439911
MeSH Arabidopsis* / genetics Arabidopsis* / metabolism Gene Editing / methods Genetic Engineering / methods Genome, Plant* Integrases / genetics Integrases / metabolism Plants, Genetically Modified Zinc Finger Nucleases / genetics Zinc Finger Nucleases / metabolism
IF 3.998
Resource
Arabidopsis / Cultured plant cells, genes rpc00008