Team:Waterloo/Project

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Chromobricks: A Platform for Chromosome Engineering with BioBricks

The aim of our project is to develop a fully-featured platform for chromosome engineering, allowing the in vivo assembly of a synthetic chromosome from interchangeable parts, followed by selective degradation of the native chromosome. We have designed a proof-of-concept for chromosome-building that will use the site-specific integrase of phage ΦC31 to integrate a BioBrick into a defined locus of the E. coli genome. Six pairs of integrase-targeted att sites have been designed to be non-cross-reactive in order to support repeatable cassette-exchange reactions for chromosome building. We have also written software to model the integrase-mediated rearrangement of DNA molecules containing att sites, to aid the design of more elaborate chromosome-building systems. To selectively degrade the native chromosome we designed a nuclease-based, inducible genome-degradation system. In its simplest form, our system can be used to integrate biological devices into a chromosome in situations requiring stable copy number and selection-free maintenance.

Project Details

Part 2

The Experiments

Cassette Exchange

BBa Donor Plasmid

Landing Pad Strain

Integrase expression plasmid

Non-crossreactive att sites

Integrase Mechanism

Design of new att sites

Design of experiment to characterize new att sites

Part 3

Results