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Team:GeorgiaTech/Project
From 2011.igem.org
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Overall project
Overuse of antibiotics in medical settings has created the problem of extremely antibiotic resistant bacteria, also called “super bugs,” which develop through the process of horizontal gene transfer of plasmids. The experiment outlined in this proposal intends to engineer a unique method of “reverse” vaccination in which the plasmid containing antibiotic resistance genes is specifically targeted and destroyed, rendering the bacteria vulnerable to antibiotic treatment. The novel method we are developing utilizes a CRISPR system (Clustered Regularly Interspaced Short Palindromic Repeats) contained on a separate plasmid to inactivate the foreign DNA. The CRISPR system is a type of immunity used by about 40% of Bacteria and about 90% of Archaea. It consists of a series of proteins, called cas proteins, which are associated with series “spacers” separated by short palindromic sequences, that recognize foreign DNA (either plasmid or viral). If foreign DNA is recognized, the cell chops up the invading DNA. If the DNA is not recognized and the cell survives the infection, it then ‘saves’ pieces of the foreign DNA in subsequent spacers, to be used in future recognition of an attack (Figure 1)
Project Details
Part 2
The Experiments
Using a modified CRISPR system that is delivered via a viral or plasmid vector, our research aims to demonstrate that antibiotic resistance genes can be targeted with the CRISPR system in multiple bacterial species, rendering the bacteria more susceptible to treatment. More specifically, artificial kanamycin resistance in a non-pathogenic S. thermophilus DGCC7710 strain will be targeted by using a CRISPR system that has a kanamycin resistance plasmid spacer. In addition, our hope is to develop computer modeling approaches to be used for understanding CRISPR spacer uptake, which is not currently well known. It is hoped with this research that new ways for targeting antibiotic resistance will be realized, helping to spark further research and innovation in this subject and CRISPR systems with real world applications.
