Development of Laser-Based Technologies and Prototype Instruments for Genome-Wide Chromatin ImmunoPrecipitation Analyses

The overall objective of the ATLAS consortium is to develop novel types of femtosecond (fs) tunable UV lasers to induce highly efficient DNA-protein crosslinking for chromatin immunoprecipitation analyses (LChIP). LChIP will overcome the current limitations of chemical crosslink. Specific, technological objectives are to fuse a microfluidic station with the ultrashort laser source in a unique, new machine to perform dynamic crosslinking in a time and cell type and/or differentiation-specific dependent manner. The LChIP technology will also be applied on frozen tissue slices for global applications. Specific biological objectives are to adapt and optimize LChIP technology to global genome analysis, i.e. on high-density tiling arrays - LChIP-chip - or massive, parallel single molecule sequencing using Solexa - LChIP-seq. Finally, we will combine Laser fixation and ChILL (Chromatin Immuno-Linked Ligation), a novel method that allows to work with low amounts of starting material. This combination technology will be developed for LChILL-chip and LChILL-seq applications.

We will further apply LChIP technologies to selected cell populations (down to the single cell level); we plan to achieve this goal i) by connecting the Laser with a microfluidic system, which may sort out specific cell populations, and ii) by selectively focusing on cell populations in solid tissues or specimens. Finally, we will develop software to automate the Laser & microfluidic station system thereby constructing a versatile and user-friendly new dedicated machine of high commercial interest.


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