Microinjection of in vitro transcribed RNA and antisense oligonucleotides in mouse oocytes and early embryos to study the gain- and loss-of-function of genes.
Mouse oocytes have proven useful in experiments aimed at studying gene function. They have been used to analyze the gain-of-function acquired after microinjection of RNA transcribed in vitro from specific gene constructs, and for establishing loss-of-function mutation obtained by injecting in vitro transcribed antisense RNA and/or synthetic oligonucleotides. This article presents protocols utilized in these studies. Specifically, the acquisition of mouse oocytes and/or embryos, the genesis of the necessary DNA and/or RNA to be used, and procedures for microinjection.
Kola
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Institute of Reproduction and Development, Monash University, Clayton, Vic., Australia. ismailko@silas.cc.monash.edu.au
Sumarsono
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D000818:Animals; D004622:Embryo, Mammalian / Q000378:metabolism*; D005786:Gene Expression Regulation; D005821:Genetic Techniques; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D008845:Microinjections; D016376:Oligonucleotides, Antisense / Q000235:genetics*; D009865:Oocytes / Q000378:metabolism*; D012333:RNA, Messenger / Q000235:genetics*
D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger