Background: We wishg to generalize our ability to measure candidate human allele functionality in zebrafish, with the sole requirements being that the gene in question is sufficiently analogous to a zebrafish gene with embryonic activity. For this we have pioneered a new platform, Nanostring, to count and correlate the quantity of injected WT and candidate mutant alleles of human RNAs with the output (activation or repression ) of a signature subset of zebrafish genes. We have applied this to two test scenarios, both in the sonic hedgehog signaling pathway: human SHH alleles and human GLI2 alleles. The results re good and we are preparing manuscripts to report upon this method and our new data.

Project Start
Project End
Budget Start
Budget End
Support Year
6
Fiscal Year
2012
Total Cost
$335,372
Indirect Cost
Name
National Human Genome Research Institute
Department
Type
DUNS #
City
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Pei, Wuhong; Kratz, Lisa E; Bernardini, Isa et al. (2010) A model of Costeff Syndrome reveals metabolic and protective functions of mitochondrial OPA3. Development 137:2587-96
Huizing, Marjan; Dorward, Heidi; Ly, Lien et al. (2010) OPA3, mutated in 3-methylglutaconic aciduria type III, encodes two transcripts targeted primarily to mitochondria. Mol Genet Metab 100:149-54
Pei, Wuhong; Feldman, Benjamin (2009) Identification of common and unique modifiers of zebrafish midline bifurcation and cyclopia. Dev Biol 326:201-11
Roessler, Erich; Pei, Wuhong; Ouspenskaia, Maia V et al. (2009) Cumulative ligand activity of NODAL mutations and modifiers are linked to human heart defects and holoprosencephaly. Mol Genet Metab 98:225-34
Domene, Sabina; Roessler, Erich; El-Jaick, Kenia B et al. (2008) Mutations in the human SIX3 gene in holoprosencephaly are loss of function. Hum Mol Genet 17:3919-28
Roessler, Erich; Ouspenskaia, Maia V; Karkera, Jayaprakash D et al. (2008) Reduced NODAL signaling strength via mutation of several pathway members including FOXH1 is linked to human heart defects and holoprosencephaly. Am J Hum Genet 83:18-29