Targeted recombination in Xenopus oocytes

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Title Targeted recombination in Xenopus oocytes
Publication Type dissertation
School or College School of Medicine
Department Biochemistry
Author Segal, David Jay
Contributor Trautman, Jon
Date 1996-12
Description The long-term goal of this project is to develop a technology that can produce genetic modifications at any position in the genome of a living cell. The central hypothesis is that the frequency of homologous recombination between an exogenous DNA and a reluctant target molecule, such as a chromosome, can be greatly enhanced by introducing a double-strand break (DSB) at the target site. The first part of the project demonstrated that a targeted DSB delivered in vivo by a site-specific endonuclease does indeed stimulate homologous recombination. The second part reported on attempts to create a more generalized in vivo, double-strand cleavage reagent. A psoralen-conjugated triplex-forming oligonucleotide was investigated for this purpose. Although psoralen is not a cleavage reagent, there were two reasons for choosing this molecule. The first was to aid in attempts to demonstrate triplex formation in Xenopus oocytes. The second was to investigate the ability of site-specific crosslinks to stimulate homologous recombination in oocytes. Unfortunately, no significant in vivo triplex formation or stimulation of recombination was observed in these experiments. These results contrast those of our collaborators, who were able to demonstrate both phenomena in COS cells using essentially the same reagents. These results suggest that a triplex based approach will be unsuccessful at enhancing the frequency of homologous recombination in oocytes, but may be useful in other cell types. Alternative approaches are discussed.
Type Text
Publisher University of Utah
Subject Genetics; Biochemistry; Crosslinking; Homologous Recobmination
Subject MESH Recombination, Genetic; Xenopus; Gene Targeting; Oocytes
Dissertation Institution University of Utah
Dissertation Name PhD
Language eng
Relation is Version of Digital reproduction of "Targeted recombination in Xenopus oocytes." Spencer S. Eccles Health Sciences Library. Print version of "Targeted recombination in Xenopus oocytes." available at J. Willard Marriott Library Special Collection. QH9.7 1996 .S43
Rights Management © David Jay Segal.
Format application/pdf
Format Medium application/pdf
Format Extent 7,846,248 bytes
Identifier undthes,4866
Source Original: University of Utah Spencer S. Eccles Health Sciences Library (no longer available).
Funding/Fellowship Grants from the National Science Foundation (MCB-9010130) and the National Institutes of Health (GM50739).
Master File Extent 7,846,305 bytes
ARK ark:/87278/s6p270wq
Setname ir_etd
ID 190771
Reference URL https://collections.lib.utah.edu/ark:/87278/s6p270wq
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