Engineering antisense oligonucleotides as a tool in synthetic biology.

• Main Author: Chng, Si Hui.
• Other Authors: Chang Wook, Matthew
• Format: Final Year Project
• Language: English
• Published: 2013
• Subjects: DRNTU::Science
• Online Access: http://hdl.handle.net/10356/52748
• Institution: Nanyang Technological University

Synthetic biology is an interdisciplinary field that comprises of the use of engineering principles to design biological systems. A distinct feature of synthetic biology is the emphasis of control over an engineered system. In this study, we used antisense oligonucleotides that did not contain any backbone modifications to repress the effects of basal transcription of an inducible promoter, acrAp. Two types of constructs, RFP and GFP were each attached to six sets of antisense oligonucleotides with different lengths. All constructs were then induced with 6 different inducer concentrations. The maximum repression level of 42.2% was achieved by a GFP construct with one-fifth GFP antisense oligonucleotides in the presence of 0.2 % (w/v) arabinose. However, results obtained were not translatable between RFP and GFP constructs. Cells with half, one-third and one-fifth RFP antisense oligonucleotides showed repression activity. In contrast, all the six GFP antisense oligonucleotides were able to lower translation level in their respective cells. This discrepancy between the two sets of results might be due to the design of the constructs. The RFP plasmid had a native RBS while RBS in GFP plasmid was inserted. Hence, this had caused a six base pair scar between the RBS and GFP coding gene.