Abstract
Abstract 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 has been developed. The targeted site in the DNA is activated by two strands of pcPNA and selectively hydrolyzed by the Ce(IV) complex. The DNA cutter required for aimed manipulation can be straightforwardly designed and obtained in terms of (i) the sequence of targeted selective scission, (ii) size of DNA substrate, and (iii) magnitude of desired site-specificity. The site-specificity can be high enough to cut the whole genome of human beings (3 × 109 base-pairs) at one predetermined site. Off-target scissions at the sites of analogous sequences hardly take place. With the use of this specificity, targeted gene can be engineered in human cells through homologous recombination. Other potential applications of this cutter, as well as recent attempts to improve its activity and specificity, are also discussed.
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