Event Title

Cloning and Expression of the DNA Binding Domain of FoxO from Ciona intestinalis That Contains an N-Terminal Nuclear Localization Signal

Poster Number

43

Presenter Information

Mikala Smith, Winthrop University

Faculty Mentor

Nicholas Grossoehme, Ph.D. and Heather Evans-Anderson, Ph.D.

College

College of Arts and Sciences

Department

Biology

Location

Richardson Ballroom

Start Date

24-4-2015 1:20 PM

End Date

24-4-2015 2:50 PM

Description

FoxO proteins are a subgroup of the Forkhead family of transcription factors. FoxO proteins are highly conserved and regulate expression of genes that control a wide variety of cellular processes, including: apoptosis, cell differentiation and proliferation, and atrophy. Ciona intestinalis is a useful model system to study developmental biology, particularly heart development, since all chordates share a conserved cardiac gene program, as well as similar cellular processes during development. Ciona FoxO (ciFoxO) protein is very similar to the FoxO1 protein in humans. In order for ciFoxO to transcriptionally regulate gene expression, it must localize to the nucleus. The major goal of this project is to add a nuclear localization signal (NLS) to an expression vector containing ciFoxO sequence that will be electroporated into Ciona embryos where it will be expressed. The N-terminal NLS will direct the exogenous ciFoxO sequence to the nucleus of cells, where it will be able to bind to target DNA sequences in the Ciona genome. The ultimate goal is to express ciFoxO constructs containing a NLS in vivo and then isolate chromatin in order to perform a ChIP-Seq assay to determine ciFoxO target genes. The ciFoxO target genes will be compared to vertebrate FoxO1 target genes to determine the level of conserved function of FoxO family members in chordates during heart development. To date, we have successfully inserted the NLS into the vector and produced dechorinated embryos; electroporation optimization is underway.

Comments

Department of Chemistry, Physics, and Geology and Department of Biology

Supported by an NIH-INBRE grant from the National Center for Research Resources and the National Institute of General Medical Sciences, an NIH grant from the National Heart, Lung, and Blood Institute and a South Carolina EPSCoR/IDeA Science Affiliate Network grant

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Apr 24th, 1:20 PM Apr 24th, 2:50 PM

Cloning and Expression of the DNA Binding Domain of FoxO from Ciona intestinalis That Contains an N-Terminal Nuclear Localization Signal

Richardson Ballroom

FoxO proteins are a subgroup of the Forkhead family of transcription factors. FoxO proteins are highly conserved and regulate expression of genes that control a wide variety of cellular processes, including: apoptosis, cell differentiation and proliferation, and atrophy. Ciona intestinalis is a useful model system to study developmental biology, particularly heart development, since all chordates share a conserved cardiac gene program, as well as similar cellular processes during development. Ciona FoxO (ciFoxO) protein is very similar to the FoxO1 protein in humans. In order for ciFoxO to transcriptionally regulate gene expression, it must localize to the nucleus. The major goal of this project is to add a nuclear localization signal (NLS) to an expression vector containing ciFoxO sequence that will be electroporated into Ciona embryos where it will be expressed. The N-terminal NLS will direct the exogenous ciFoxO sequence to the nucleus of cells, where it will be able to bind to target DNA sequences in the Ciona genome. The ultimate goal is to express ciFoxO constructs containing a NLS in vivo and then isolate chromatin in order to perform a ChIP-Seq assay to determine ciFoxO target genes. The ciFoxO target genes will be compared to vertebrate FoxO1 target genes to determine the level of conserved function of FoxO family members in chordates during heart development. To date, we have successfully inserted the NLS into the vector and produced dechorinated embryos; electroporation optimization is underway.