A simple, efficient way to introduce large plasmid DNA into cells using functional peptides

April 25, 2019

A joint research team from RIKEN CSRS and Rikkyo University have successfully used a functional peptide to introduce large plasmid DNA into Escherichia coli cells more efficiently and easily than conventional methods, with less damage.

Conventional electroporation method introduces large plasmid DNA into cells by opening pores in the cell membrane with electric pulses. However, when the electric pulse is too strong, it can cause significant cell damage. Electroporation therefore has the disadvantage of requiring optimization of conditions according to cell types.

The joint research team used a functional fusion peptide composed of a cell-penetrating peptide and a polycationic peptide, and mixed it with a 205 kb plasmid DNA, which is much larger than the conventional plasmid (~10 kb), to form peptide-DNA complex. Using the peptide-DNA complex, the large plasmid DNA was successfully introduced into the E. coli cells with an efficiency comparable to conventional electroporation, and the plasmid DNA was less damaged and degraded. The functional protein was produced from the reporter gene in the introduced plasmid.

These findings are expected to contribute to gene cluster transformation to produce materials in microorganisms and to the introduction of chromosomal DNA into cells to create artificial cells.

 

Original article
ACS Synthetic Biology doi:10.1021/acssynbio.9b00055
M. M. Islam, M. Odahara, T. Yoshizumi, K. Oikawa, M. Kimura, M. Su’etsugu, K. Numata,
"Cell-penetrating peptide-mediated transformation of large plasmid DNA into Escherichia coli".

Contact
Masaki Odahara; Research Scientist
Keiji Numata; Team Leader
Biomacromolecules Research Team