Identifier	Thesis 2143
Author	Pozo, Pedro Nicolas,1984-
Title	Discovery of RNA editing in the grin1b transcipt of zebrafish (Danio rerio): a predictive and experimental approach
Publisher	Central Connecticut State University;
Date of Publication	2011
Resource Type	Master's Thesis
Abstract	Discovering RNA editing sites in model organisms provides an insight into their adaptations in addition to finding potential sites for gene expression regulation. In this study, we focused on the grin1b gene of Zebrafish due to its important function in the nervous tissue as a glutamate receptor ion channel. We used NCBI computer programs to locate 5 possible RNA-editing events within the grin1b transcript and designed and ordered primers targeted at these possible editing sites. One primer pair, F3R3, gave us a robust band at the size predicted which upon sequencing identified E5 as a potential editing site; this was verified by DNA controls. Surprisingly, sequence analysis also revealed a new editing site, E6, which was not observed in the predictive computer models. By comparing these cDNA sequences to the genomic sequence of grin1b as a control, we show for the first time the discovery of two novel RNA editing events in the grin1b transcript of Zebrafish. Studying the frequency levels of these events with the help of restriction enzymes and densitometry analyses enabled us to further characterize this novel finding. Editing at site E5 created a restriction enzyme site for the endonuclease MluI, whereas editing at site E6 created a restriction enzyme site for the endonuclease BstNI. Upon completion of restriction digests using these enzymes and by using densitometry analyses we found the frequency of editing at site E5 to be 26.98% and site E6 to be 21.36%. Collectively, this study shows for the first time the discovery of RNA editing in the grin1b gene transcript of Zebrafish. This exciting finding represents the first time RNA editing is observed in this glutamate receptor ion channel and provides further insight into this molecular process. Moreover, by characterizing this finding in terms of frequency and location we hope to advance our current knowledge of RNA editing with the ultimate goal of elucidating this intriguing molecular process.
Subject	RNA editing Zebra danio
Department	Department of Biomolecular Sciences
Advisor	Hoopengardner, Barry
Type	Text;
Digital Format	application/pdf
Language	eng
OCLC number	804652989
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Discovery of RNA editing in the grin1b transcipt of zebrafish (Danio rerio): a predictive and experimental approach

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