Construction and Characterization of Gene Regulatory Networks in Yeast

• Main Author: Jedrysiak, Daniel K.
• Language: en
• Published: 2013
• Subjects: gene; regulatory; network; synthetic biology; yeast; DNA; cloning; BrickMason; GRN; CYC8; TUP1; SSN6
• Online Access: http://hdl.handle.net/10393/24000

Two major roadblocks in synthetic biology are the difficulties associated with the physical assembly of gene regulatory networks (GRNs) and the lack of characterized biological parts. In this work we aimed to address both of these issues. We developed a novel method for the assembly of GRNs called Brick- Mason assembly. We have shown that the method can assemble a 6 part network in a single day and provides significant advancements over traditional cloning methods. We used BrickMason to assemble GRNs that would allow us to compare natural yeast mechanisms of repression to the steric hindrance based mechanisms that are commonly used in synthetic GRNs in yeast. Our results show that the two mechansisms of repression are not equivalent. This finding opens possibilities for using a new class of repressor in a synthetic context in yeast.