Identification and Characterization of Novel FMRP-Associated miRNAs
Abstract
Project rationale and original aims: New evidence suggests that Fragile X Mental Retardation Protein (FMRP) may need to associate with ancillary factors to co-regulate the production of some of important synaptic proteins. Currently, there is a critical need to identify and characterize these molecules. Some factors believed to associate with FMRP and co-regulate target mRNAs include a group of small regulatory molecules called microRNAs (miRNAs). There are two long-term goals associated with this project. First, to use an established model system for FXS to identify and functionally characterize all FMRP-associated miRNAs involved in the control of synapse structure and function. Second, to identify and functionally characterize all mRNA targets for these miRNAs. Progress towards completion of aims: First, after setbacks developing novel transgenic epitope-tagged FMRP protein that works efficiently in our assays, we have adapted two established antibody-based approaches to sequence FMRP-associated miRNAs and mRNAs: 1) direct co-immunoprecipitation of FMRP-containing RNPs (FMRP-IP); and 2) high-throughput sequencing of RNA isolated by crosslinking immunoprecipitation (HITS-CLIP). The latter will allow us to identify FMRP-associated mRNAs and specific FMRP binding sites. Second, a serendipitous discovery as allowed us to make substantial progress towards understanding the molecular mechanisms involved in FMRP-mediated recruitment of miRNAs to target mRNAs. This will allow us to very rapidly and efficiently validate bona fide target mRNAs.
Document Details
- Document Type
- Technical Report
- Publication Date
- Oct 01, 2013
- Accession Number
- ADA606025
Entities
People
- Scott A. Barbee