Nichols A, Rungger-Brändle E, Muster L, Rungger D
Station de Zoologie expérimentale, University of Geneva, Chêne-Bougeries, Switzerland.
Mech Dev. 1995 Jul;52(1):37-49. doi: 10.1016/0925-4773(95)00387-g.
Antisense inhibition of gene expression during Xenopus development was obtained by injecting, into the zygote, an expression vector carrying the adenovirus VAI gene read by RNA polymerase III. This vector yields high levels of antisense RNA in most embryonic cells between mid-blastula transition and tailbud stage. As a target we chose the Xenopus homeobox gene Xhox1A. A 26 bp long oligonucleotide, including the initiation codon of this gene, was inserted in opposite polarity into the vector. Antisense treatment reduces Xhox1A mRNA in embryos up to stage 22 and Xhox1A protein expression up to stage 30. Half of the antisense-treated embryos develop a characteristic phenotype with disorganized somites in the anterior trunk and delayed development of the intestinal tract.
通过将携带由RNA聚合酶III转录的腺病毒VAI基因的表达载体注射到非洲爪蟾的受精卵中，实现了对非洲爪蟾发育过程中基因表达的反义抑制。该载体在囊胚中期转换至尾芽期之间的大多数胚胎细胞中产生高水平的反义RNA。作为靶标，我们选择了非洲爪蟾同源框基因Xhox1A。将一段包含该基因起始密码子的26 bp长寡核苷酸以相反极性插入载体中。反义处理可使胚胎中直至22期的Xhox1A mRNA以及直至30期的Xhox1A蛋白表达减少。经反义处理的胚胎中有一半会出现特征性表型，即躯干前部的体节排列紊乱以及肠道发育延迟。
