A method using electroporation for the protein delivery of Cre recombinase into cultured Arabidopsis cells with an intact cell wall View Full Text


Ontology type: schema:ScholarlyArticle      Open Access: True


Article Info

DATE

2019-12

AUTHORS

Yuichi Furuhata, Ayako Sakai, Tomi Murakami, Mone Morikawa, Chikashi Nakamura, Takeshi Yoshizumi, Ushio Fujikura, Keiji Nishida, Yoshio Kato

ABSTRACT

Genome engineering in plants is highly dependent on the availability of effective molecular techniques. Despite vast quantities of research, genome engineering in plants is still limited in terms of gene delivery, which requires the use of infectious bacteria or harsh conditions owing to the difficulty delivering biomaterial into plant cells through the cell wall. Here, we describe a method that uses electroporation-mediated protein delivery into cultured Arabidopsis thaliana cells possessing an intact cell wall, and demonstrate Cre-mediated site-specific recombination. By optimizing conditions for the electric pulse, protein concentration, and electroporation buffer, we were able to achieve efficient and less-toxic protein delivery into Arabidopsis thaliana cells with 83% efficiency despite the cell wall. To the best of our knowledge, this is the first report demonstrating the electroporation-mediated protein delivery of Cre recombinase to achieve nucleic acid-free genome engineering in plant cells possessing an intact cell wall. More... »

PAGES

2163

References to SciGraph publications

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  • Identifiers

    URI

    http://scigraph.springernature.com/pub.10.1038/s41598-018-38119-9

    DOI

    http://dx.doi.org/10.1038/s41598-018-38119-9

    DIMENSIONS

    https://app.dimensions.ai/details/publication/pub.1112165529

    PUBMED

    https://www.ncbi.nlm.nih.gov/pubmed/30770845


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