PhAST Technology

The team of Dynamics of Macromolecular Complexes at the LBPA has developped  an  approach  to use the probability of UV induced cyclobutane dimer formation between adjacent pyrimidines on the same DNA chain in naked and bound DNA as a probe of changes in local base structure not only between naked DNA and reconstituted nucleosomes but also at different stages of nucleosome formation. This technique of Photo-chemical Analysis of Structural Transitions (PhAST) induces photo-chemical alterations in DNA pyrimidine dimer formation. The location and intensities of these alterations are themselves affected by external agents that reshape the DNA structure and thus alter the photo-chemistry. Thus, the technique provides unique information on the local structure at a base level of the DNA. We reveal photochemical products using a simple primer extension technique coupled to capillary electrophoresis. Our approach confers high-resolution, excellent quantification, application in vitro and in vivo, and the possibility of high-throughput since practically any size DNA sample may be analysed in a fashion analogous to genome sequencing.

Concerning nucleosomes, a recent result, which can be found here: 2016 Hatakeyama demonstrated that, for the first time we are able to follow dynamic changes at a molecular level in DNA during nucleosome assembly and disassembly.

The technique has also been applied to liquid crystal DNA as shown here: 2018 Brach.

 

 

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