SURF 2015: Rapid prototyping of moderate complexity biomolecular circuits

From MurrayWiki
Jump to: navigation, search

2015 SURF project description

  • Mentor: Richard M. Murray
  • Co-mentors: Clare Hayes and Vipul Singhal
Breadboards-process.png
Overview of the cell-free expression breadboard process.

The overall goal of this project is to build a set of “biomolecular breadboards” to create a systematic, engineering-oriented approach to synthesizing biomolecular circuits that involves developing, modeling, and debugging a sequence of prototype devices, each at increasing levels of complexity and each allowing the incorporation of increasingly realistic operating environments for either in vitro or in vivo applications. We are adapting an existing cell-free toolbox developed at U. Minnesota to create a set of prototyping environments for testing biological circuits. A sequence of increasingly complex environments, ending in prokaryotic cells, is being used to demonstrate the ability to prototype circuits that function in vivo, with iteration in in vitro assays and models to streamline development of predictable, in vivo functionality.

This SURF project will focus on the design and implementation of a modest complexity (8-12 promoter) circuit using the TX-TL breadboard that has been previous developed. Students will select a circuit during the SURF proposal process. During the summer, the design will proceed in several (partially overlapping) phases:

  1. The circuit will be modeled using an existing TX-TL toolbox (built on top of MATLAB) and this model will be used to verify the basic dynamics of the circuit, as well as used to test hypotheses as the circuit is implemented and debugged.
  2. Individual components of the circuit will be built and tested using the TX-TL experimental breadboard. Experimental iterations typically take 1-2 days per iteration, with up to 50 variations tested per cycle. Measurements will be taken using fluorescent reporters and a plate reader.
  3. Once component operation is verified (experimentally and using mathematical models), the components will be composed in the TX-TL toolbox and adjusted as needed to demonstrate functionality.
  4. Components, combinations of components and the full system will also be tested in vivo, but transforming the circuits into lab strains of E. coli and measured using either a plate reader or a flow cytometer.

Required skills: Students interested in this project should have some basic laboratory skills (equivalent to Bi 1X, Bi 10 or ChE/BE 130) and have experience with either MATLAB, Python or a similar scientific programming package.

References