A venturesome research project is proposed to develop the tools needed to create transgenic quail embryos. The techniques of transgenesis in the mouse embryo are largely responsible for the dramatic recent progress in the molecular genetics of vertebrate development. Avian embryos are selected because they are accessible to study at all stages of development without the heroic efforts that are required to study mouse embryos. Because of these advantages, the major system for classical work in the developmental and cell biology of higher vertebrates was the chicken. Here we propose to begin the refinement of molecular genetic tools that are heretofore only available in the mouse onto the quail embryo. The quail offers advantages in the small size of its egg, the moderate size of the breeding adults, and its short generation time. Because of these advantages, the proposed work will allow molecular genetic experiments on a higher vertebrate embryo both more rapidly and less expensively than comparable work on the mouse. Our overriding goal is to combine transgenic quail technology with the use of two-photon laser-scanning microscopy, so that transgenes containing the coding sequence for the green fluorescent protein (GFP) can be used to follow gene expression patterns in living embryos. We will use HIV based vectors, which have been shown to efficiently generate murine transgenics without being silenced and maintaining endogenous gene expression, to insert the GFP expressing transgenes within the egg of the developing avian embryo. Although ambitious, the project builds from ongoing work in the laboratory on avian development, optimized viral vectors for vertebrate embryos and quantitative microscopy of living avian embryos. These ongoing efforts provide the fertile ground needed for the transgenic avian project to succeed. ? ?

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21HD047347-02
Application #
7229946
Study Section
Development - 1 Study Section (DEV)
Program Officer
Coulombe, James N
Project Start
2006-04-01
Project End
2008-03-31
Budget Start
2007-04-01
Budget End
2008-03-31
Support Year
2
Fiscal Year
2007
Total Cost
$160,480
Indirect Cost
Name
California Institute of Technology
Department
Type
Schools of Arts and Sciences
DUNS #
009584210
City
Pasadena
State
CA
Country
United States
Zip Code
91125
Anderson, Maxwell R; Akeeb, Ameenat; Lavela, Joseph et al. (2017) Period 3 gene polymorphism and sleep adaptation to stressful urban environments. J Sleep Res 26:115-118
Huss, David; Benazeraf, Bertrand; Wallingford, Allison et al. (2015) A transgenic quail model that enables dynamic imaging of amniote embryogenesis. Development 142:2850-9
Seidl, Armin H; Sanchez, Jason Tait; Schecterson, Leslayann et al. (2013) Transgenic quail as a model for research in the avian nervous system: a comparative study of the auditory brainstem. J Comp Neurol 521:5-23
Sato, Yuki; Poynter, Greg; Huss, David et al. (2010) Dynamic analysis of vascular morphogenesis using transgenic quail embryos. PLoS One 5:e12674
Cui, Cheng; Cheuvront, Tracey J; Lansford, Rusty D et al. (2009) Dynamic positional fate map of the primary heart-forming region. Dev Biol 332:212-22
Poynter, Greg; Huss, David; Lansford, Rusty (2009) Japanese quail: an efficient animal model for the production of transgenic avians. Cold Spring Harb Protoc 2009:pdb.emo112
Poynter, Greg; Huss, David; Lansford, Rusty (2009) Injection of lentivirus into stage-X blastoderm for the production of transgenic quail. Cold Spring Harb Protoc 2009:pdb.prot5118
Poynter, Greg; Huss, David; Lansford, Rusty (2009) Screening for transgenic Japanese quail offspring. Cold Spring Harb Protoc 2009:pdb.prot5119
Poynter, Greg; Huss, David; Lansford, Rusty (2009) Generation of high-titer lentivirus for the production of transgenic quail. Cold Spring Harb Protoc 2009:pdb.prot5117
Huss, David; Poynter, Greg; Lansford, Rusty (2008) Japanese quail (Coturnix japonica) as a laboratory animal model. Lab Anim (NY) 37:513-9

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