Header Logo
Last Name

You can now add alternative names! Click here to add other names that you've published under.

Technology for Identification of Epiproteomes in Tissue Samples

Collapse Overview 
Collapse abstract
? DESCRIPTION (provided by applicant): The overall goal of this proposal is to develop a cutting-edge technology for identifying epiproteomes in tissue samples, which will provide an approach that will redefine how NIDA researchers study epigenetic responses to drug abuse/addiction. We define an epiproteome as the proteins, histones and histone posttranslational modifications at a defined ~1 kb chromosomal location. In previous work, we have developed a groundbreaking technology platform called ChAP-MS (Chromatin Affinity Purification with Mass Spectrometry) that allows us to biochemically isolate a ~1 kb section of a chromosome and identify the epiproteome using high resolution proteomics. Our ChAP-MS platform provided for the first ever isolation of a specific, native chromatin locus for proteomic analysis. The first and second generation ChAP-MS technologies used ectopic expression of affinity probes for chromatin isolation and were designed for cell culture studies; thus, precluding the analysis of animal models. As outlined in this proposal, we will develop a third generation approach called recombinant rCRISPR-ChAP-MS in yeast and translate it to brain tissue samples. The third generation approach will circumvent ectopic expression of affinity probes; thus, enabling the analysis of tissue. We envision the long term application of the rCRISPR-ChAP-MS approach in defining epigenetic mechanisms regulating drug abuse/addiction. Specific Aim 1 will focus on developing rCRISPR-ChAP-MS for application to tissue samples. Using S. cerevisiae as a model system in Aim 1, we will develop rCRISPR-ChAP-MS by analysis of epiproteomes at the GAL1 promoter under transcriptionally active and repressed conditions. Specific Aim 2 will focus on translating rCRISPR-ChAP-MS to brain tissue. In Aim 2, we will target our optimized approach to the gene promoter of AMPA receptor subunit GluA1 in striatum from rats chronically exposed to methamphetamine.

Collapse sponsor award id

Collapse Biography 

Collapse Time 
Collapse start date

Collapse end date