Compartmentalized Self-Tagging for In Vitro-Directed Evolution of XNA Polymerases.

Template-dependent synthesis of xenobiotic nucleic acids (XNAs) is an essential step for the development of functional XNA molecules, as it enables Darwinian evolution to be carried out with novel genetic polymers. The extraordinary substrate specificity of natural DNA polymerases greatly restricts the spectrum of XNAs available, thus making it necessary to identify DNA polymerase variants capable of incorporating a wider range of substrates. This unit summarizes compartmentalized self-tagging (CST), a directed evolution strategy developed for the selection of DNA polymerase variants capable of XNA synthesis.