ABSTRACT
Understanding of RNA interference (RNAi) has been made possible through a variety of experimental approaches using different model organisms [1]. Some of these systems are amenable to mutational screening, facilitating forward genetic approaches and subsequent discovery of novel genes involved in the RNAi pathway. Others permit biochemical approaches producing mechanistic insight. Over the last 8 years, observations from different groups using these techniques in different systems have yielded a clear outline of the basic mechanisms behind RNAi [2]. These approaches have also uncovered a new class of genes that encode microRNAs (miRNAs), which
Overview ................................................................................................................ 149 9.1 Introduction to Zebra Fish ............................................................................ 150
9.1.1 Characteristics of Zebra Fish and Embryonic Development ............ 150 9.1.2 Zebra Fish as an Experimental System ............................................ 150
9.2 RNAi Technology ......................................................................................... 152 9.2.1 RNAi in Different Model Systems ................................................... 152 9.2.2 Application of RNAi in Zebra Fish .................................................. 153 9.2.3 Small Hairpin RNAs and RNAi in Zebra Fish ................................ 154
9.3 The Vertebrate siRNA/miRNA Pathway ..................................................... 155 9.3.1 Zebra Fish RNAi Components ......................................................... 156 9.3.2 Conservation of RNAi Components ................................................. 156
9.4 Zebra Fish Dicer Mutants ............................................................................. 158 9.4.1 Recovering a Dicer1 Null Allele ....................................................... 158 9.4.2 Eliminating All Dicer Function in Zebra Fish Embryos .................. 161 9.4.3 Implications of the Dicer Null Phenotype ........................................ 162
9.5 Zebra Fish miRNAs ...................................................................................... 163 9.5.1 Dicer Mutants Reveal a Role for the miR-430 Family ..................... 163 9.5.2 miRNA Diversity in Zebra Fish ....................................................... 164 9.5.3 Strategies for Determining miRNA Function in Zebra Fish ............ 165
9.6 Conclusions ................................................................................................... 166 References .............................................................................................................. 168
act through an RNAi-like mechanism to regulate gene expression. Here, we will discuss advances in RNAi/miRNA research using the zebra fish model system, and discuss how this system may help answer outstanding questions in the field.
