Collaborative Research: Repurposing the translation apparatus for mirror image polypeptide synthesis

Project: Research project

Project Details

Description

Project Overview: The translation system (the ribosome and associated factors) is the cell’s factory for protein synthesis. The extraordinary capacity of the protein synthesis machinery has driven extensive efforts to harness it for novel functions. As an example, genetic code expansion for biosynthesis of proteins containing non-standard amino acids has elucidated new biological understanding and opened the way to novel applications in medicine, energy, and materials that are difficult, if not impos¬sible, to achieve by other methods.

This work sets the stage to use engineered translation systems to manufacture and study novel materials. Concurrently, we will create an educational and outreach program integrated with the proposed research for teaching and training 21st century scientists and engineers in synthetic biology.

Intellectual Merit: The proposed work aspires to catalyze a new paradigm for engineering translation using genome engineering technologies. The level of control afforded by cell-free systems and their potential advantages are significant. First, they may open the way to evolving OTS systems with higher catalytic efficiency than exists currently. Second, they facilitate development of OTSs in scenarios wherein nsAAs cannot permeate the cell or the OTS components are deleterious to evolutionary fitness. Third, they provide a unique opportunity to expand the genetic code in conditions of reduced system complexity when structural barriers are absent. Just as synthetic organic chemistry once revolutionized the ability of chemists to build molecules following a basic set of design rules (including those that did not exist in nature), the work proposed will deliver an invaluable toolbox for harnessing and expanding the chemistry of life. Such a technology platform will be instrumental in engineering more diverse OTSs and alternate genetic codes. In addition, our ability to study and control phosphorylation outside the restrictive confines of a cell will help answer fundamental questions such as how PTMs influence protein properties and function. From an engineering perspective, our research will make possible the production of therapeutic phosphoproteins, greatly expanding the scope of cell-free systems and synthetic biology.

Broader Impacts: Our new paradigm for building designer translation systems will advance fundamental understanding of protein synthesis, provide a general platform to efficiently encode many chemically diverse nsAAs, and enable the biosynthesis of homogeneous phosphoproteins containing multiple distinct nsAAs. The ability to produce useful quantities of proteins featuring multiple user-specified phosphorylations will be instrumental in opening new areas of study that draw conclusive linkages between a specific PTM state and protein activity (e.g., elucidating the histone code and human kinome). We will also implement an integrated plan to promote education and discourse in synthetic biology and science more generally for K12, undergraduate, and graduate students, and next-generation community leaders. A key focus is the expansion of STEM education and career opportunities for underrepresented minorities and women. We will create experiential learning modules that bring cell-free synthetic biology research to classrooms and provide students with real-world experience of the science conducted as part of this CAREER proposal. We will also promote public engagement by fostering a community of early career practitioners. This education program will ensure that advances made in this project bene
StatusActive
Effective start/end date9/1/178/31/21

Funding

  • National Science Foundation (MCB-1716766)

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