CRISPR-Cas systems are recently discovered, RNA-based immune systems that control invasions of viruses and plasmids in prokaryotes. Prokaryotes with CRISPR-Cas systems capture short invader sequences within the CRISPR loci in their genomes, and small RNAs produced from the CRISPR loci (CRISPR (cr)RNAs) are thought to guide Cas proteins to recognize and silence the invading nucleic acids;however, we know very little about how the key steps in the remarkable CRISPR-Cas immune response pathways occur. In this project, we address the molecular basis for the three steps required for CRISPR-Cas defense through the following specific aims: ? Determine how CRISPR loci acquire foreign DNA sequences ? Determine the mechanisms by which functional CRISPR RNAs are produced ? Delineate the mechanisms of invader silencing Specifically, we aim to obtain a molecular understanding of the CRISPR-Cas defense pathways that function in Streptococcus thermophilus. We will identify the essential cellular machinery (proteins and RNAs) and molecular mechanisms involved in each of the three key phases of CRISPR-Cas defense. We hypothesize that Cas proteins function in each of the major steps in the defense pathway, and we will test this and a number of specific related hypotheses in the proposed studies. The experiments will define the CRISPR-Cas pathways used by multitudes of prokaryotes to survive viral attack. The information gained in this study will contribute directly o ongoing efforts aimed at exploiting CRISPR-Cas systems to both strengthen domesticated bacteria (used to produce safe food, pharmaceuticals and biofuels) and weaken human pathogens and limit the spread of antibiotic resistance.

Public Health Relevance

Scientists recently recognized that bacteria depend on a unique immune system to defend themselves against viruses. Our studies will define the major steps and essential factors in this defense pathway. We are very interested in studying this pathway because the discoveries that we make have tremendous potential to be used to protect bacteria that we depend on to make food products, pharmaceuticals, and biofuels, and to combat bacteria that cause human disease and the spread of antibiotic resistance.

Agency
National Institute of Health (NIH)
Institute
National Institute of General Medical Sciences (NIGMS)
Type
Research Project (R01)
Project #
5R01GM099876-02
Application #
8531996
Study Section
Molecular Genetics B Study Section (MGB)
Program Officer
Bender, Michael T
Project Start
2012-09-01
Project End
2016-04-30
Budget Start
2013-05-01
Budget End
2014-04-30
Support Year
2
Fiscal Year
2013
Total Cost
$272,275
Indirect Cost
$88,925
Name
University of Georgia
Department
Biochemistry
Type
Schools of Arts and Sciences
DUNS #
004315578
City
Athens
State
GA
Country
United States
Zip Code
30602
Terns, Rebecca M; Terns, Michael P (2014) CRISPR-based technologies: prokaryotic defense weapons repurposed. Trends Genet 30:111-8
Carte, Jason; Christopher, Ross T; Smith, Justin T et al. (2014) The three major types of CRISPR-Cas systems function independently in CRISPR RNA biogenesis in Streptococcus thermophilus. Mol Microbiol 93:98-112