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Degree

PHD, Kent State University, College of Biomedical Sciences, 2008.

Abstract

NF-kB plays a central role in several physiological processes such as immunity, inflammation, development and cell survival. Under normal conditions, NF-kB activation is, subjected to several layers of regulation, check-points and feed-back controls and hence is rapid and transient in nature. Elevated NF-kB activity is a hallmark of various autoimmune and inflammatory diseases. Moreover, it is now well established that constitutive NF-kB activation is strongly associated with origination, maintenance, progression and therapeutic resistance of cancer. Deciphering how constitutive activation of NF-kB is achieved in cancer cells is now an arena of intense research. However, there still remain considerable gaps in our understanding of pathological perturbations of one of the most intensely studied signaling networks. This gap has been a natural target for forward genetics. Forward genetics plays an important role in revealing the details of complex signaling pathways. This method involves choosing a specific pathway of interest, designing appropriate screening or selection methods for isolation of mutants defective in that pathway and employing methods of complementation or reversion that allow identification of the genetic locus that has been altered in the mutant. We have used the synthetic, functional transposon Sleeping Beauty (SB) for random insertional mutagenesis. An outbound promoter embedded in the transposon facilitates production of dominant mutants and permits regulation of their phenotypes. The SB system was used to randomly mutagenize C6TA4 cells, which harbor a system for selection of mutants with constitutive NF-kB activity. Our studies have identified a previously unrecognized short form of the receptor-interacting protein kinase RIP1 as a factor capable of constitutively up-regulating NF-kB.

Subject Headings

Biology, Molecular

Keywords

NF-kB; RIP1; Transposons; TNF; Insertional Mutagenesis

Advisor

George R Stark

Pages

140p.

Document number: kent1201807856
Permalink: http://rave.ohiolink.edu/etdc/view?acc_num=kent1201807856

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