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Method Article
Yoshio Kato
Yoshio Kato Lab
Corresponding Author
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Cre recombinase is widely used in a variety of organisms to manipulate DNA both in vitro and in vivo. Combined with its consensus DNA sequences, named loxP, it allows the DNA modification including deletions, insertions, translocations and inversions at specific DNA locus of cells. To promote its use in plant cultured cells, we report here a detailed protocol for direct protein delivery of Cre recombinase into cultured Arabidopsis thaliana cells with an intact cell wall. Electroporation with the optimized buffer and program enables the efficient delivery of Cre protein and following recombination in cultured Arabidopsis thaliana cells. Specifically, we describe the purification of Cre protein, the preparation of electroporation samples, and the assessment of electroporated cells. This simple, economical, and effective technology will contribute to the biological analysis of genes and cellular functions in cultured Arabidopsis thaliana cells. This protocol accompanies Furuhata et al (DOI: 10.1038/s41598-018-38119-9), Scientific Reports, published online on February 15th 2019.
Keywords
Genome engineering, Cre recombinase, Electroporation, Protein delivery, Arabidopsis thaliana, Cell wall
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Associated Publications
A method using electroporation for the protein delivery of Cre recombinase into cultured Arabidopsis cells with an intact cell wall
by Yuichi Furuhata, Ayako Sakai, Tomi Murakami, +6
Scientific Reports (11 March, 2019)
An open repository of community-contributed protocols sponsored by Nature Research.
Learn more about Protocol Exchange
Method Article
Yoshio Kato
Yoshio Kato Lab
Corresponding Author
Version History
CURRENT STATUS: Posted
Version 1
Posted 14 Mar, 2019
No community comments so far
Metrics
Comments: 0
HTML Views: ...
License:
This work is licensed under a CC BY 4.0 License. Read Full License
Cre recombinase is widely used in a variety of organisms to manipulate DNA both in vitro and in vivo. Combined with its consensus DNA sequences, named loxP, it allows the DNA modification including deletions, insertions, translocations and inversions at specific DNA locus of cells. To promote its use in plant cultured cells, we report here a detailed protocol for direct protein delivery of Cre recombinase into cultured Arabidopsis thaliana cells with an intact cell wall. Electroporation with the optimized buffer and program enables the efficient delivery of Cre protein and following recombination in cultured Arabidopsis thaliana cells. Specifically, we describe the purification of Cre protein, the preparation of electroporation samples, and the assessment of electroporated cells. This simple, economical, and effective technology will contribute to the biological analysis of genes and cellular functions in cultured Arabidopsis thaliana cells. This protocol accompanies Furuhata et al (DOI: 10.1038/s41598-018-38119-9), Scientific Reports, published online on February 15th 2019.
Figure 1
Figure 2
None.
Loading...
Associated Publications
A method using electroporation for the protein delivery of Cre recombinase into cultured Arabidopsis cells with an intact cell wall
by Yuichi Furuhata, Ayako Sakai, Tomi Murakami, +6
Scientific Reports (11 March, 2019)