RRC ID 3937
Author Nishiyama A, Fujiwara S.
Title RNA interference by expressing short hairpin RNA in the Ciona intestinalis embryo.
Journal Dev. Growth Differ.
Abstract We carried out RNA interference by expressing short hairpin RNA (shRNA) in the Ciona intestinalis embryo. For this purpose, we identified a gene encoding U6 small nuclear RNA (snRNA) in the C. intestinalis genome. The 1-kb sequence upstream of the U6 snRNA gene was sufficient for directing transcription of short RNA as revealed by Northern blot hybridization. An shRNA-expressing plasmid vector was constructed, in which shRNA-encoding oligonucleotides are inserted downstream of the U6 promoter. An shRNA that contained a sequence homologous to the C. intestinalis tyrosinase gene (Ci-tyrosinase) suppressed melanization of pigment cells in the brain of morphologically normal tailbud embryos. An shRNA that perfectly matched the translated sequence of enhanced green fluorescent protein (EGFP) (a mutant type of Aequorea victoria green fluorescent protein) suppressed the expression of the coelectroporated EGFP transgene. These results suggest that the expression of shRNA interferes with functions of both endogenous and exogenous genes. The shRNA-expressing plasmid constructed in the present study provides an easy and inexpensive alternative for the functional analysis of genes in ascidian embryos.
Volume 50(6)
Pages 521-9
Published 2008-8
DOI 10.1111/j.1440-169X.2008.01039.x
PII DGD1039
PMID 18510713
MeSH Animals Ciona intestinalis / embryology* Ciona intestinalis / genetics* Gene Expression Gene Expression Regulation, Developmental* Gene Silencing Genes, Reporter Green Fluorescent Proteins / metabolism Monophenol Monooxygenase / biosynthesis Monophenol Monooxygenase / genetics Promoter Regions, Genetic RNA / metabolism* RNA Interference Transgenes
IF 1.496
Times Cited 15
WOS Category DEVELOPMENTAL BIOLOGY CELL BIOLOGY
Resource
C.intestinalis / (O.japonicus) Wild C.int