期刊
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
卷 -, 期 118, 页码 -出版社
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/54972
关键词
Genetics; Issue 118; peptide-based delivery; organelle targeting; gene expression; plasmid DNA; double-stranded DNA; double-stranded RNA; protein; Arabidopsis thaliana; tobacco
资金
- Japan Science and Technology Agency Exploratory Research for Advanced Technology (JSTERATO)
- New Energy and Industrial Technology Development Organization (NEDO)
- Cross-ministerial Strategic Innovation Promotion Program (SIP), Japan
The capacity to introduce exogenous proteins and express (or down-regulate) specific genes in plants provides a powerful tool for fundamental research as well as new applications in the field of plant biotechnology. Viable methods that currently exist for protein or gene transfer into plant cells, namely Agrobacterium and microprojectile bombardment, have disadvantages of low transformation frequency, limited host range, or a high cost of equipment and microcarriers. The following protocol outlines a simple and versatile method, which employs rationally-designed peptides as delivery agents for a variety of nucleic acid-and protein-based cargoes into plants. Peptides are selected as tools for development of the system due to their biodegradability, reduced size, diverse and tunable properties as well as the ability to gain intracellular/ organellar access. The preparation, characterization and application of optimized formulations for each type of the wide range of delivered cargoes (plasmid DNA, double-stranded DNA or RNA, and protein) are described. Critical steps within the protocol, possible modifications and existing limitations of the method are also discussed.
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