Silence of the fish: injection of photoswitchable short interfering RNA oligonucleotides into Japanese medaka embryos (oryzias latipes) to photochemically control gene silencing

The siRNA duplex functions by binding to and cleaving mRNA, a process known as the RNAi pathway. Existing siRNAs face challenges such as off-target effects and unpredictable prolonged gene-silencing. To address these issues, enhancing their therapeutic potential, siRNAs can be modified controlling their role in the RNAi pathway. An Ortho-functionalized, tetrafluorinatedazobenzene was integrated into the siRNA backbone via phophoramidite chemistry to generate a class of photoswitchable F-siRNAs. These F-siRNAs retained the ability to photoisomerize from an active trans to an inactive cis state through blue and green light respectively. This thesis reported on a novel technique to inject single cell medaka embryos with both Wt and F-siRNAs. Medaka are a small freshwater teleost fish that have a number of desirable features for use as a vertebrate model in gene silencing projects. These attributes include daily spawning, a number of useful genetic strains, a completely sequenced genome, and a transparent egg.

Name thesis:degree_name

Master of Science (MSc)

Discipline thesis:degree_discipline

Applied Bioscience

Grantor

University of Ontario Institute of Technology

Year dc:date.issued

2023

Author dc:creator

• Mateus, Makenzie

Advisors dc:contributor.advisor

• Desaulniers, Jean-Paul
• Simmons, Denina

• siRNA
• Antisense
• Japanese medaka
• Nano injection
• Transgenic

Language dc:language.iso

en

Handle dc:identifier.uri

https://hdl.handle.net/10155/1727

OAI identifier oai:identifier

oai:ontariotechu.scholaris.ca:10155/1727