IGEM:IMPERIAL/2007/CFS

Cell Free Systems

• Introduction
• Specifications
• Design
• Modelling
• Implementation
• Testing/Validation
• Notes
• References

Introduction

• What is a chassis
• Introduce CFS as in vitro gene expression machinery systems

Components of CFS

• Types of Extracts
  • E. coli extracts, ie. S30, S12
  • Bacteriophage expression E.coli extracts, ie. T7, SP6
  • Wheatgerm extracts
  • Japanese

• Types of Compartmentalization
  • Bulk Solution
  • Semi-continuous
  • Continuous
  • Vesicles (phospholipids)

Chassis Comparison Table

• Characteristics that we talked earlier + stuff from earlier characterization attempt

Advantages

Disadvantages

Achievements

Cell Free Extracts

• Attempted to make S30 E. coli cell free extracts using French Press.
• Successfully used commercial S30 extracts from Promega for our experiments.
• Successfully made pre-reaction mixture and implemented it in commcercial cell extracts.

Vesicles

• Successfully implemented the 'Mineral Oil' Method of vesicle formation.
• Successfully implemented empty vesicle formation and vesicles encapsulating GFP.
• Attempting to enclose cell extracts in vesicles to induce gene expression.
• Attempting to find suitable pore proteins to prolong vesicle and expression lifespan