RRC ID |
54599
|
著者 |
Furuhata Y, Sakai A, Murakami T, Morikawa M, Nakamura C, Yoshizumi T, Fujikura U, Nishida K, Kato Y.
|
タイトル |
A method using electroporation for the protein delivery of Cre recombinase into cultured Arabidopsis cells with an intact cell wall.
|
ジャーナル |
Sci Rep
|
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.
|
巻・号 |
9(1)
|
ページ |
2163
|
公開日 |
2019-2-15
|
DOI |
10.1038/s41598-018-38119-9
|
PII |
10.1038/s41598-018-38119-9
|
PMID |
30770845
|
PMC |
PMC6377677
|
MeSH |
Arabidopsis / metabolism
Arabidopsis / radiation effects*
Cell Wall / metabolism
Cell Wall / radiation effects*
Electroporation / methods*
Endocytosis*
Integrases / metabolism*
Plant Cells / metabolism
Plant Cells / radiation effects*
Protein Transport*
|
IF |
4.011
|
引用数 |
4
|
リソース情報 |
シロイヌナズナ / 植物培養細胞・遺伝子 |
rpc00008 |