Functional knockdown of neuropilin-1 in the developing chick nervous system by siRNA hairpins phenocopies genetic ablation in the mouse.

摘要: The chick embryo is widely used for the study of vertebrate development, but a general, reliable loss-of-function strategy analysis gene function currently not available. By using small inhibitory hairpin RNA (siRNA) molecules generated by mouse U6 promoter, we have applied an interference approach to achieve quantitative knockdown neuropilin-1 (Nrp-1) receptor in embryos. Functional was evident abolition Sema3A-induced growth cone collapse Nrp-1-siRNA Nrp-2-siRNA-expressing dorsal root ganglion (DRG) neurons. Two nervous system defects Nrp-1 mutant mice were phenocopied embryos treated with siRNA. First, DRG axons prematurely entered horn and projected inappropriately. Second, targeted early migrating neural crest cells destined sympathetic chain arrested ectopically within ventral spinal nerve roots. Localized induced specific siRNA constructs will allow rapid functional genes regulating development whilst circumventing embryonic lethal effects often associated global knockout mouse.