Improvement of E. coli transcription-translation (TX-TL) system

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This project is focuses on the improving methods of production of cell-free transcription-translation (TX-TL) systems. It is funded by the Caltech Grubstake program, which is aimed at transitioning Caltech research to commercial implementation.

Primary participants:

Additional participants:


Collaborators:
  • Synvitrobio, Inc.

Past participants:

  • Clare Hayes (research tech)
  • Alex White (, )
  • Tiffany Zhou (undergraduate)
  • Zachary Sun (PhD student, Biology)
  • Abel Chiao (research technician)

Overview

Breadboards-process.png
Overview of the cell-free expression breadboard process.

In vitro E. coli lysate systems have been used for more than a half-century to probe biological phenomena. However, the advancement of molecular and synthetic biology tools has resulted in increased alternative applications. In particular, in vitro systems emulate a simplistic cellular environment for rapid biological circuit prototyping. In vitro systems can also produce large amounts of protein in a controlled manner. Despite recent application advancements, there has not been commiserate research into lysate protocols. As a result, lysate development has been costly and not tuned to the specific application. We have developed a novel in vitro transcription-translation system, or TX-TL, which has shown high demand from collaborators outstripping supply. We believe that that we can increase applicability and decrease production costs by 2-5X, enabling viable commercialization of the TX-TL system.

Project aims:

  • Optimize preparation methods for TX-TL to meet application needs. Current preparation methods to make extract for circuit prototyping are low-yield (18 mL per batch). However, alternative preparation methods exist (45 mL per batch) which are significantly less labor-intensive, but are not optimized for circuit prototyping. Research on preparation methods will be conducted to increase yields but match cellular conditions more precisely.
  • Reduce costs of production of TX-TL by 2-5X. Current costs, including labor, are $0.03/µL. However, with preparation modifications as well as adjustments of reaction formulas we believe we can decrease this cost. In particular, labor constitutes 60% of the cost, but the current TX-TL protocol is relatively inefficient.

Publications

None to date



  • Agency: Caltech Grubstake
  • Grant number:
  • Start date: 30 March 2015
  • End date: 31 Aug 2016
  • Support: ~1 technician + supplies
  • Reporting: final report due at end of project