Site-selective scission of human genome by artificial restriction DNA cutter.

  title={Site-selective scission of human genome by artificial restriction DNA cutter.},
  author={Kenichiro Ito and Hitoshi Katada and Narumi Shigi and Makoto Komiyama},
  journal={Chemical communications},
By using an artificial restriction DNA cutter which is composed of Ce(iv)/EDTA and two pseudo-complementary peptide nucleic acid strands (pcPNAs), only one target site in the whole genome of human beings (one site in the X chromosome) was selectively hydrolyzed. 

Design and Applications of Artificial Restriction DNA Cutters for Site-Selective Scission of Genomes

By combining Ce(IV)/EDTA complex and pseudo-complementary peptide nucleic acid (pcPNA), an artificial restriction DNA cutter which hydrolyzes targeted phosphodiester linkages in double-stranded DNA...

Site-selective scission of human genome using PNA-based artificial restriction DNA cutter.

This chapter describes the method of site-selective scission of human genomic DNA using ARCUT, a completely chemistry-based artificial restriction DNA cutter by combining a pair of pseudo-complementary PNA strands and Ce(IV)/EDTA complex (molecular scissors).

Artificial site-selective DNA cutters to manipulate single-stranded DNA

With the cutters composed of two oligonucleotide additives and Ce(IV)/EDTA, long single-stranded DNA can be selectively cut at target site and manipulated according to the needs.

Site-specific Manipulation of Mitochondrial DNA by Artificial Restriction DNA Cutter

Site-specific gene manipulation using homologous recombination induced by a double-strand break in DNA is expected to be applied to mitochondrial genome manipulation. In this study, by using a chem...

Clipping of predetermined fragments from the human genome by S1 nuclease-PNA combinations.

By combining S1 nuclease with two strands of pseudo-complementary peptide nucleic acid (pcPNA), the whole human genome was selectively cut at targeted sites, and desired fragments were clipped from

Site-selective Scission of Double-stranded DNA by Combining a Triplex-forming bis-PNA and Ce(IV)/EDTA

By combining triplex-forming bis-peptide nucleic acid (bis-PNA) with Ce(IV)/EDTA, double-stranded DNA was selectively hydrolyzed at a predetermined homopyrimidine/homopurine site, which is difficul...

Artificial Restriction DNA Cutters ( ARCUT ) as New Tools to Manipulate Human Genome

Artificial restriction DNA cutter (ARCUT) which cuts double-stranded DNA at one desired site using Ce(IV)/EDTA complex as molecular scissors and a pair of pseudo-complementary PNAs, and their scission-site is determined by simple Watson-Crick rule.

Artificial DNA cutters for DNA manipulation and genome engineering.

This tutorial review provides recent developments in artificial cutters for site-selective scission of DNA with the focus on chemistry-based DNA cutters, since their site- selectivity is much higher than that of naturally occurring restriction enzymes and also their scission site is freely chosen.

Site-selective DNA hydrolysis induced by a metal-free peptide nucleic acid-cyclen conjugate.

A metal-free artificial restriction DNA cutter which is composed of cyclen and classical peptide nucleic acid (PNA) was synthesized. Analysis of DNA cleavage products indicates the site-selective

Chemical modifications of artificial restriction DNA cutter (ARCUT) to promote its in vivo and in vitro applications

  • M. Komiyama
  • Biology, Chemistry
    Artificial DNA, PNA & XNA
  • 2014
Strong potential ofpcPNA for further applications in vivo and in vitro has been confirmed and new strategies using conventional PNA, in place of pcPNA, as site-selective activator are presented.



Artificial Restriction DNA Cutters as New Tools for Gene Manipulation

Two types of artificial tools that cut double‐stranded DNA through hydrolysis of target phosphodiester linkages, have been recently developed.

Artificial restriction DNA cutter for site-selective scission of double-stranded DNA with tunable scission site and specificity

The artificial restriction DNA cutter (ARCUT) method to cut double-stranded DNA at designated sites has been developed and the location of the scission site and the site-specificity are almost freely tunable, and there is no limitation to the size of DNA substrate.

Chemical modification of Ce(IV)/EDTA-based artificial restriction DNA cutter for versatile manipulation of double-stranded DNA

Potentiality of ARCUT for manipulation of huge DNA has been substantiated by site-selective scission of genomic DNA of Escherichia coli (composed of 4,600,000 bp) at the target site.

Origin of high fidelity in target-sequence recognition by PNA-Ce(IV)/EDTA combinations as site-selective DNA cutters.

Mismatches in (or near) the central double-invasion region are especially fatal, showing that Watson-Crick pairings of the DNA bases in this region with the pcPNA strands are essential for precise recognition of the target sequence.

Site-specific cleavage of human chromosome 4 mediated by triple-helix formation.

This method may facilitate the orchestrated dissection of human chromosomes from normal and affected individuals into megabase sized fragments and facilitate the isolation of candidate gene loci.

Hybrid restriction enzymes: zinc finger fusions to Fok I cleavage domain.

The deliberate creation of novel site-specific endonucleases by linking two different zinc finger proteins to the cleavage domain of Fok I endonuclease and the zinc finger motifs opens the way to generate many new enzymes with tailor-made sequence specificities desirable for various applications.

Double duplex invasion by peptide nucleic acid: a general principle for sequence-specific targeting of double-stranded DNA.

  • J. LohseO. DahlP. Nielsen
  • Biology, Chemistry
    Proceedings of the National Academy of Sciences of the United States of America
  • 1999
It is predicted that (for decameric targets) more than 80% of all sequences can be targeted by straightforward Watson-Crick base pairing by using this approach in its present form.

A novel engineered meganuclease induces homologous recombination in yeast and mammalian cells.

Two novel, chimeric meganucleases, derived from homing endonucleases are presented, able to induce recombination in yeast and mammalian cells, whereas the second cleaves a novel DNA target site.

Site-selective and hydrolytic two-strand scission of double-stranded DNA using Ce(IV)/EDTA and pseudo-complementary PNA.

By combining Ce(IV)/EDTA with two pseudo-complementary peptide nucleic acids (pcPNAs), both strands in double-stranded DNA were site- selectively hydrolyzed at the target site, resulting in the site-selective two-strand scission of the double-Stranded DNA.

Chemical‐Reaction‐Based Site‐Selective DNA Cutter for PCR‐Free Gene Manipulation

An artificial restriction DNA cutter (ARCUT), recently developed by the authors, was used to construct a fusion protein that was successfully expressed in mammalian cells, and showed entirely different subcellular localization from EGFP itself.