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Site-Selective Scission of Human Genome Using PNA-Based Artificial Restriction DNA Cutter

  • Kenichiro Ito
  • Makoto Komiyama
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1050)

Abstract

Site-selective scission of genomes is quite important for future biotechnology. However, naturally occurring restriction enzymes cut these huge DNAs at too many sites and cannot be used for this purpose. Recently, we have developed a completely chemistry-based artificial restriction DNA cutter (ARCUT) by combining a pair of pseudo-complementary PNA (pcPNA) strands (sequence recognition moiety) and Ce(IV)/EDTA complex (molecular scissors). The scission site of ARCUT and its scission specificity can be freely modulated in terms of the sequences and lengths of the pcPNA strands so that even huge genomes can be selectively cut at only one predetermined site. In this chapter, the method of site-selective scission of human genomic DNA using ARCUT is described in detail.

Key words

Artificial restriction DNA cutter Site-selective scission Human genome Pseudo-complementary PNA (pcPNA) Ce(IV)/EDTA complex 

Notes

Acknowledgements

This work was partially supported by Grants-in-Aid for Specially Promoted Research from the Ministry of Education, Science, Sports, Culture, and Technology, Japan (18001001 and 22000007) and by the Global COE Program for Chemistry Innovation.

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Copyright information

© Springer Science+Business Media, New York 2014

Authors and Affiliations

  • Kenichiro Ito
    • 1
  • Makoto Komiyama
    • 1
    • 2
  1. 1.Research Center for Advanced Science and TechnologyThe University of TokyoTokyoJapan
  2. 2.Life Science Center of Tsukuba Advanced Research AllianceUniversity of TsukubaTsukubaJapan

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