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Graduate Seminar Series: Molecular Stream – Teddi Shaw

December 2, 2022 @ 12:00 pm - 12:30 pm EST

Graduate Seminar Series: Molecular Stream
Graduate Seminar Series for the Institute of Biomedical Engineering (BME). This day is for molecular stream presenters.
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Presentation Title: Improving AAV targeting through nanobody inclusion by alternative splicing.
Abstract: Adeno-associated viruses (AAVs) are viral vectors that can be engineered to act as a gene therapy. AAVs contain a single-stranded DNA genome, with a capsid protein shell. Although AAVs have desirable characteristics for a gene therapy, their targeting and specificity can be improved. To increase the specificity of AAV targeting, the inclusion of nanobodies into the surface of the AAV capsid is proposed. This concept will be proven in plasmids by synthetically developing alternative splicing, specifically, intron retention. This results in intron inclusion in the final protein. In this construct, the nanobody will become the intron so, we can optimize how many copies of the nanobody are included in the final product. To alter the strength of the splicing, various properties will be modified. Firstly, linker sequences that flank the intron differentiate the intron-exon border and promote splicing of the intron. The 5’ linker sequence contains the splice donor and the 3’ linker contains the splice acceptor, polypyrimidine tract (PPT) and the branch point. These features promote the assembly of the spliceosome, thereby increasing splicing activity. Secondly, G triplets located in the interior of an intron enforce intron-exon borders. To investigate splicing activity, three constructs are proposed. The first is read-through, or no splicing, where both the linker sequences and G triplets are absent. In the weak splicing construct, only linker sequences are present. Finally, in the strong splicing construct, linker sequences and G triplets are present. When intronic elements are included in the construct, the splice percentage is increased, thereby lowering the inclusion of the intron. Once this methodology is proven in plasmids, it will be applied to AAVs.
Supervisor Name: Dr. Kevin Truong
Year of Study: 2
Program of Study: MASc
Zoom link: https://us02web.zoom.us/j/89610372821?pwd=azd4SCtYVWtreVovaGNPV1c2NGY2Zz09
Meeting ID: 896 1037 2821
Password: 483329
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December 2, 2022
12:00 pm - 12:30 pm EST
Event Category: