Photoassisted Recovery of DNA Molecules for On-chip Directed Evolution

Abstract

Directed evolution is a powerful approach to creating functional biomolecules, and a combinative use of microarray chip technology is expected to have a great potential for realizing a sophisticated directed evolution with higher speed and quantitative performance than ever before. Although “recovery” of evaluated and selected molecules followed by amplification is an essential step in the performance of directed evolution, conventional DNA or protein microarray chips are limited only to analytical use. In this study, we have developed a novel technology for DNA microarray that is applicable to an artificial Darwinian selection on a chip. DNA molecules were microarrayed on a gold surface via an oligonucleotide linker that contains a nitrobenzyl group as a photocleavable moiety and were recovered from the surface by spot-selective photoirradiation at few micrometer precision. The recovered DNA molecules were confirmed by polymerase chain reaction. Thus, in this study, we have realized a new function of biomolecular microarray technology that is useful for the establishment of on-chip directed molecular evolution.