Chemically modified DNA (M-DNA) possesses several useful properties, including expanded reactivity and nuclease resistance. However, native DNA polymerases cannot synthesize M-DNA strands with the length or accuracy required for many of the applications that would make it useful. Significant progress has been made in recent years towards generating high-fidelity M-DNA polymerases with sufficient activity for use in processes such as PCR. While these have demonstrated increased processivity and lower error rates, accuracy still needs to be increased by orders of magnitude to approach natural error rates and make M-DNA polymerases useful for applications that require high fidelity. In this project, we have added mutations that were shown to increase the fidelity of wild-type Taq polymerases to the best current M-DNA polymerases. We then thoroughly characterized the fidelity of these new M-DNA polymerases, and found improvements in fidelity relative to their parent enzymes as well as other changes that were similar to previous findings in wild-type Taq polymerases.

Improving M-DNA polymerase accuracy through rational design

ACS Spring 2022

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.