Investigation of the RNA recognition machinery involved in dendritic targeting of prominent denritically localized mRNAs.
To elucidate the RNA recognition mechanism in transport RNPs I am using different affinity purification methods:
- through aptamer-tagged RNAs and
- through double-tagged (already known) protein factors
to isolate RNA-protein complexes (neuronal RNPs) from rat brain lysates
(in vitro approach) and transiently transfected neurons (in vivo approach).
These approaches allow the analysis of protein-protein and protein-RNA interactions specific for a given RNA localization element, and the specificity of interactions can be also studied this way by using different mutant RNAs for binding assays.
The following questions are addressed in my project:
- What RNA sequences (localization elements) and RNA binding proteins (RBPs) are involved in dendritic localization of a specific transcript?
- What RNA sequences and RBPs are shared in dendritic localization of different transcripts?
- Are there conserved core complexes of RBPs in localized neuronal RNPs (core RNPs)?
- Which non-coding RNAs are involved in the regulation of dendritic localization of a specific transcript?