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Nanomaterials for Gene Knockdown In Vivo

Stenkamp Laboratory

The zebrafish is an outstanding model system for the study of the developmental genetics of organogenesis and cell differentiation. Our goal is to determine whether nanomaterials can be utilized as carriers of antisense oligonucleotides in order to achieve temporally- and/or spatially-selective gene knockdown in embryonic zebrafish. The gene knockdown methods in use (morpholino-conjugated oligonucleotides = “morpholinos”) do not offer this flexibility – they must be delivered very early in development, and they continue to be active through the period of organogenesis. Because many developmentally important genes are deployed at multiple times, and in multiple locations, a more spatiotemporally-selective strategy will be the key to understanding the tissue-specific functions of these developmental genes.

We have been conducting preliminary studies to test for toxicity of nanomaterials, and to determine the distribution of oligonucleotides delivered with nanomaterials. Our project will address the following Specific Aims:
Aim 1. Evaluate toxicity of nanomaterials during embryonic development in vivo, both with and without associated oligonucleotides.
Aim 2. Evaluate distribution of oligonucleotides delivered to embryos using nanomaterials.
Aim 3. Assess effectiveness of gene knockdown following nanomaterial-based delivery of oligonucleotides.