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Abstract

CRISPR (clustered regularly interspaced short palindromic repeat) arrays and Cas(CRISPR-associated) proteins provide bacteria and archaea with immunity against phages and other mobile genetic elements (MGEs). The immunity provided by CRISPR-Cas systems is adaptive as sequences are acquired from invaders and stored in the CRISPR array, capable of guiding sequence-specific nuclease activity during future encounters. In response, phages and other MGEs encode anti-CRISPR proteins that inhibit the defense of CRISPR-Cas immunity. This dissertation begins with characterization of CRISPR-Cas systems in Streptococcus thermophilus with an emphasis on Type III-A systems. It then explores anti-CRISPR protein prediction in phages of S. thermophilus followed by screening and identification of novel CRISPR-Cas inhibitors. The final chapter focuses on the selectivity of spacer targeting against S. thermophilus phage genes with potential implications for our understanding of Type III-A CRISPR-Cas system function. The results increase predictability of outcomes of phage-host encounters by expanding the repertoire of known anti-CRISPR proteins and illuminating unique features and potential roles of cooccurring CRISPR-Cas systems.

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