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Structural and Functional Characterization of T.thermophilus CasE Open Access


Other title
CRISPR, RNA, Prokaryotes, gene expression regulation, endonuclease
Type of item
Degree grantor
University of Alberta
Author or creator
Gesner, Emily
Supervisor and department
MacMillan, Andrew (Department of Biochemistry)
Examining committee member and department
Bleackley, R Chris (Department of Biochemistry)
MacMillan, Andrew (Department of Biochemistry)
Glover, M (Department of Biochemistry)
Unrau, P (Molecular Biology and Biochemistry, SFU)
Owttrim, G (Department of Biological Sciences)
Department of Biochemistry

Date accepted
Graduation date
Doctor of Philosophy
Degree level
Powerful mechanisms of genetic interference in both unicellular and multicellular organisms are based on the sequence-directed targeting of DNA or RNA by small effector RNAs. In many bacteria and almost all archaea, RNAs derived from clustered, regularly interspaced, short palindromic repeat (CRISPR) loci are involved in an adaptable and heritable gene-silencing pathway. Resistance to phage infection is conferred by the incorporation of short invading DNA sequences into the prokaryotic genome as CRISPR spacer elements separated by short repeat sequences. A central aspect to this pathway is the processing of a long primary transcript (pre-crRNA) containing these repeats by crRNA endonucleases to generate the mature effector RNAs that interfere with phage or plasmid gene expression. Here we describe a structural and functional analysis of the CasE endonuclease of T. thermophilus a member of the Ecoli CRISPR sub-type. High resolution X-ray structures of CasE bound to repeat RNAs model both the pre-and post-cleavage complexes associated with processing the pre-crRNA. These structures establish the molecular basis of a specific CRISPR RNA recognition and suggest the mechanism for generation of effector RNAs responsible for gene-silencing.
License granted by Emily Gesner ( on 2011-04-05T02:45:23Z (GMT): Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of the above terms. The author reserves all other publication and other rights in association with the copyright in the thesis, and except as herein provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.
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