Morpholinos: Studying gene function in the chick

被引:10
作者
Norris, Anneliese [1 ]
Streit, Andrea [1 ]
机构
[1] Kings Coll London, Dept Craniofacial Dev & Stem Cell Biol, London SE1 9RT, England
基金
英国生物技术与生命科学研究理事会;
关键词
Avian embryo; Gene knockdown; In vivo electroporation; Loss-of-function; OVO ELECTROPORATION; IN-VIVO; NEURAL-TUBE; EXPRESSION; EMBRYOS; INHIBITION; KNOCKDOWN; SYSTEM; OLIGOS; DNA;
D O I
10.1016/j.ymeth.2013.10.009
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The use of morpholinos for perturbing gene function in the chick, Gallus gallus, has led to many important discoveries in developmental biology. This technology makes use of in vivo electroporation, which allows gain and loss of function in a temporally, and spatially controlled manner. Using this method, morpholinos can be transfected into embryonic tissues from early to late developmental stages. In this article, we describe the methods currently used in our laboratory to knock down gene function using morpholinos in vivo. We also detail how morpholinos are used to provide consistency of the results, and describe two protocols to visualise the morpholino after electroporation. In addition, we provide guidance on avoiding potential pitfalls, and suggestions for troubleshooting solutions. These revised techniques provide a practical starting point for investigating gene function in the chick. (C) 2013 Elsevier Inc. All rights reserved.
引用
收藏
页码:454 / 465
页数:12
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