Poster abstracts

Poster number 13 submitted by Nicole Wagner

DNA structure and dynamics in site-specific recombination

Nicole Wagner (OSBP)

Abstract:
Cre Recombinase catalyzes site-specific DNA recombination at pseudo-palindromic loxP sites through two rounds of strand cleavage, exchange, and religation. Cre possesses unmet potential as a gene editing tool due its lack of requirements for external energy sources or host factors, as well as the fact that its recombination products do not require repair of double-stranded DNA breaks. However, broader applications of Cre in editing noncanonical target sequences requires a deeper understanding of the DNA features that enable target site selection and efficient recombination. Although Cre recombines loxP DNA in a specific and ordered fashion, it makes few direct contacts to the loxP spacer, the region where recombination occurs. We are using NMR to test the hypothesis that sequence-specific dynamic and structural features of the loxP spacer enable recognition by Cre. We report resonance assignments and analysis of flanking sequence effects and mutations on spacer chemical shifts. These results set the stage for NMR-based structure determination and dynamics measurements.

Keywords: DNA, NMR, recombination