Whiplash PCR-based methods of biomolecular computation (BMC), while highly-versatile in principle, are well-known to suffer from a simple but serious form of self-poisoning known as back-hybridization. In this work, an optimally re-engineered WPCR-based architecture, Displacement Whiplash PCR (DWPCR) is proposed and experimentally validated. DWPCR's new rule protect biostep, which is based on the primer-targeted strand-displacement of backhybridized hairpins, renders the most recently implemented rule-block of each strand unavailable, abolishing back-hybridization after each round of extension. In addition to attaining a near-ideal efficiency, DWPCR's ability to support isothermal operation at physiological temperatures eliminates the need for thermal cycling, and opens the door for potential biological applications. DWPCR should also be capable of supporting programmable exon shuffling, allowing XWPCR, a proposed method for programmable protein evolution, to more closely imi...
John A. Rose, Ken Komiya, Satsuki Yaegashi, Masami