Informace o projektu
Development of novel in-cell NMR experiments and data interpretation schemes for ab initio determination of nucleic acid structure

Here, we propose a pilot study aiming towards development of conceptually novel suite of in-cell NMR experiments and data interpretation schemes for ab initio determination of nucleic acid (NA) structure in a complex environment of living cells. In contrast to conventional methods for NA determination in vitro, which are predominantly based on NOE derived distances, the proposed scheme will exploit residual dipolar couplings and residual chemical shift anisotropies resulting from inherent self-alignment of NA in strong magnetic field. The scheme will be complemented with conventional NMR data, such as 13C and 31P chemical shifts, 1JC-H, and C-HC, reporting on local NA structure. The proposed approach aims to overcome limitations imposed on acquisition-time of in-cell NMR data and their spectral resolution due to limited in vivo stability of the NA as well as the survival time of the cells in the NMR tube and inherent inhomogeneity of in-cell NMR sample, respectively.