Gene interference using antisense oligodeoxynucleotides on whole chick embryos. Optimal ring and roller-bottle culture technique.
Details are given of an advanced version of the ring method of chick embryo culture. This ensures good development from early blastoderm stages even when the culturing procedure is interrupted by the extended periods required for collecting matched embryo samples and for preparing antisense treatment. Antisense oligodeoxynucleotide treatment, in short-term incubation before return of blastoderms to their ring cultures, is then described. An alternative, roller-bottle, culture method for continued development after treatment is also described. Criteria for the validity and success of this gene interference method are given. While the text is meant to be of detailed practical help to those inexperienced in embryo culture, a preliminary reading, and familiarity with its sectional (Subheading) structure, is recommended before work is undertaken.
Cooke
J
J
National Institute for Medical Research, London, UK.
Isaac
A
A
D000818:Animals; D001756:Blastoderm / Q000166:cytology / Q000502:physiology*; D002642:Chick Embryo; D046508:Culture Techniques / Q000295:instrumentation / Q000379:methods*; D018390:Gene Targeting; D018014:Gene Transfer Techniques; D008954:Models, Biological; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D013873:Thionucleotides / Q000494:pharmacology*
D016376:Oligonucleotides, Antisense; D013873:Thionucleotides