Cell‐Free Protein Synthesis: A Platform Technology for Education

Abstract

Hands‐on learning of transcription and translation remains limited due to a lack of classroom modules for early‐career students. Currently, classrooms implement animations, simulations, games, and bacterial transformation/protein expression to teach the genetic code. The cell wall poses the primary barrier for students to directly observe and manipulate the intracellular environment, limiting inquiry‐based learning of the genetic code. We have overcome this limit by levering transcription and translation in vitro. The open nature of cell‐free protein synthesis (CFPS) is achieved by removing the cell wall and genomic DNA, while isolating the cellular machinery involved in transcription and translation. This allows experimenter to harness the genetic code in a test tube and directly manipulate the reaction conditions for hands‐on learning. While reconstitution of cellular machinery in vitro for CFPS has been transformative for biotechnology, CFPS has not been accessible to early‐career students due to the complexity in reagent preparation and reaction set‐up. We have successfully adapted the CFPS biotechnology to develop a learning module accessible to early‐career STEM students focusing on learning objectives involving the genetic code. Here we report the reformulation of the CFPS reaction setup for simplicity and improved reagent shelf life in order to adapt CFPS for the classroom. First, our simplified platform requires the pipetting of only 3 pre‐mixed reagents, an 80% reduction in setup steps, making it more accessible to an early‐career student with little‐to‐no laboratory experience. Second, our simplified platform is shelf‐stable at −20°C rather than −80°C, making it accessible for storage and implementation in undergraduate and high school classrooms. Longer‐term experiments are in progress to validate shelf‐life at 4°C and room temperature. Our data show that these improvements are achieved without sacrificing CFPS performance in reaction kinetics or total protein yield. Third, our analysis of green fluorescent protein synthesis kinetics shows that 90% of maximum protein yields can be achieved within 3 hours at 37°C, making this platform compatible with most college laboratory courses. We believe that this educational technology will be transformative to the way the genetic code is taught in classrooms today. Toward this end, we have developed a CFPS classroom ‘kit’ for broad dissemination.Support or Funding InformationOur research is funded by the Center for Applications in Biotechnology @ Cal Poly SLO, Bill and Linda Frost Fund, and NSF‐1708919.This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.