Cut-and-Paste of DNA Using an Artificial Restriction DNA Cutter

• Main Author: Makoto Komiyama
• Format: Article
• Language: English
• Published: MDPI AG 2013-02-01
• Series: International Journal of Molecular Sciences
• Subjects: site-selective scission; DNA cutter; Ce(IV)/EDTA; PNA; human genome; homologous recombination; restriction enzyme-free manipulation
• Online Access: http://www.mdpi.com/1422-0067/14/2/3343

DNA manipulations using a completely chemistry-based DNA cutter (ARCUT) have been reviewed. This cutter, recently developed by the authors, is composed of Ce(IV)/EDTA complex and two strands of pseudo-complementary peptide nucleic acid. The site-selective scission proceeds via hydrolysis of targeted phosphodiester linkages, so that the resultant scission fragments can be easily ligated with other fragments by using DNA ligase. Importantly, scission-site and site-specificity of the cutter are freely tuned in terms of the Watson–Crick rule. Thus, when one should like to manipulate DNA according to the need, he or she does not have to think about (1) whether appropriate “restriction enzyme sites” exist near the manipulation site and (2) whether the site-specificity of the restriction enzymes, if any, are sufficient to cut only the aimed position without chopping the DNA at non-targeted sites. Even the human genome can be manipulated, since ARCUT can cut the genome at only one predetermined site. Furthermore, the cutter is useful to promote homologous recombination in human cells, converting a site to desired sequence. The ARCUT-based DNA manipulation should be promising for versatile applications.