Poster abstracts

Poster number 46 submitted by Munira Fouz

Enhanced tools for manipulating and analyzing gene function in a sea star larval model to explore regenerative biology

Munira Fouz (Department of Chemistry, Carnegie Mellon University), Lauren Xu (Department of Chemistry, Carnegie Mellon University), Veronica Hinman (Department of Bioloical Sciences, Carnegie Mellon University), Danith Ly (Department of Chemistry, Carnegie Mellon University), Bruce Armitage (Department of Chemistry, Carnegie Mellon University)

Abstract:
Humans have little capacity to regenerate. However, human tissues could potentially be reengineered if the mechanisms controlling this process in animals that do regenerate are properly understood. While vertebrate animals and humans in particular have generally poor regenerative capabilities, members of their closest invertebrate phylum, sea stars, have extraordinary abilities to regenerate. The sea star larva is a newly emerged model system for understanding the regulatory mechanisms of cell specification during early development. Here we develop enhanced fluorescent detection tools using gamma modified Peptide Nucleic Acid (PNA) mini probes, to assay small RNA expression and to perturb gene function. These mini probes will be powerful detection and signal amplification agents for Whole Mount InSitu Hybridization (WMISH) to visualize spatial patterns of microRNA expression. The melting curves of the ternary duplexes formed by the mini probes and a synthetic analog of a representative micro RNA show highly cooperative affinity towards the target and a significant destabilization with a single mismatch RNA sequence; an important feature to discriminate among closely related miRNA family members. These tools are likely to be widely useful for any developmental model system.

References:
Goldman, J. M.; Zhang, L. A.; Manna, A.; Armitage, B. a; Ly, D. H.; Schneider, J. W. High Affinity γPNA Sandwich Hybridization Assay for Rapid Detection of Short Nucleic Acid Targets with Single Mismatch Discrimination. Biomacromolecules 2013, 14, 2253–2261.

Keywords: Regenerative biology, Peptide Nucleic Acids (PNA), Whole Mount InSitu Hybridization (WMISH)