Functional genomics requires reliable methods to investigate the consequences of disrupting the gene function. Since its discovery in C .elegans, RNA interference (RNAi) has become an invaluable resource to tackle gene function in cell culture, mammals and invertebrate model systems. However, but it has proven a major challenge to apply this technique in zebrafish and Xenopus. In this proposal we will apply our current knowledge in the microRNA (miRNA) field to develop several strategies that improve RNAi. This proposal will provide specific rules to the scientific community to improve RNAi in zebrafish to allow the functional characterization of the zebrafish genes. In the future, the resuls derived from this project will establish a frame work to understand the structure and the function of vertebrate genes.

Public Health Relevance

Understanding the function of genes requires reliable methods to block gene expression. Over the last decade, it has been discovered that double strand RNA can be used to shut down expression of genes (RNA interference) however this technique is not available in zebrafish. This proposal aims to understand the steps required to apply this powerful technique to zebrafish, what will help us understand the function of vertebrate genes in development and disease.

Agency
National Institute of Health (NIH)
Institute
Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD)
Type
Exploratory/Developmental Grants (R21)
Project #
5R21HD073768-02
Application #
8501614
Study Section
Development - 2 Study Section (DEV2)
Program Officer
Coulombe, James N
Project Start
2012-07-01
Project End
2014-06-30
Budget Start
2013-07-01
Budget End
2014-06-30
Support Year
2
Fiscal Year
2013
Total Cost
$237,013
Indirect Cost
$94,663
Name
Yale University
Department
Genetics
Type
Schools of Medicine
DUNS #
043207562
City
New Haven
State
CT
Country
United States
Zip Code
06520
Lee, Miler T; Bonneau, Ashley R; Giraldez, Antonio J (2014) Zygotic genome activation during the maternal-to-zygotic transition. Annu Rev Cell Dev Biol 30:581-613
Yoda, Mayuko; Cifuentes, Daniel; Izumi, Natsuko et al. (2013) Poly(A)-specific ribonuclease mediates 3'-end trimming of Argonaute2-cleaved precursor microRNAs. Cell Rep 5:715-26