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MuDLOC Résumé de rapport

Project ID: 309600
Financé au titre de: FP7-IDEAS-ERC
Pays: Israel

Mid-Term Report Summary - MUDLOC (Multi-Dimensional Lab-On-Chip)

MuDLOC is a multi-dimensional microfluidic based platform for proteomics. The core technology of MuDLOC is a DNA to protein array. This means that a DNA array is turned into a protein by in vitro transcription and translation inside the device, at nanoliter volumes.
The advantages of this approach are many. For example, there is no need to purify proteins. The method is also compatible with all protein types including, as we demonstrate, on thousands of membrane proteins. We also demonstrate that the fresh proteins produced on the MuDLOC platform are mostly functional. This is a significant improvement over most previous protein arrays, in which protein functionality was a serious issue.
Measuring interactions on protein arrays is performed in non-equilibrium conditions. This leads to loss of information and adversely affects the sensitivity of these methods. By using integrated microfluidics and applying a MITOMI, we are able to measure the interactions at equilibrium. This led directly to higher sensitivity and for example for discovery of “weak” or transient interactions.
In the past 2 years, we have worked in several parallel avenues: We have invested a lot of energy on improving the fabrication process and the core platform. We have bioengineered new assays that will serve as additional layers or dimensions to the platform. These include, DNA methylation, protein post translation modifications (starting with phosphorylation and ubiquitination) and tools for characterizing protein DNA binding. The latter is very important for understanding gene regulation.
The final goal in MuDLOC is a multi-dimensional platform that can study and characterize protein networks in vitro. All on a single platform, including protein-protein, protein-DNA and DNA or protein modifications. For example, we expect to be able to look at a DNA binding protein and discover its protein regulator (by interaction or modification). Next, on the same platform and same conditions, we can characterize the effect of these regulations on the DNA binding properties of the protein. This may lead to better and more accurate models predicting where and when the protein will bind on our genome.

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