Fu Y, Wang Y, Evans S M
Department of Medicine, University of California, San Diego, La Jolla 92093-0613, USA.
Nat Biotechnol. 1998 Mar;16(3):253-7. doi: 10.1038/nbt0398-253.
Expression of transgenes within a single generation by direct DNA injection into vertebrate embryos has been plagued by inefficient and nonuniform gene expression. We report a novel strategy for efficient and stable expression of transgenes driven by both ubiquitous and tissue-specific promoters by direct DNA injection into developing Xenopus laevis embryos. This strategy involves flanking expression cassettes of interest with inverted terminal repeat sequences (ITRs) from adeno-associated virus. Our results suggest that the ITR strategy may be generally applicable to other systems, such as zebra fish and embryonic stem cells, and may enable tissue-specific expression of transgenes in problematic contexts.
通过将DNA直接注射到脊椎动物胚胎中，在单一世代内实现转基因表达一直受到基因表达效率低下和不均匀的困扰。我们报告了一种新策略，通过将DNA直接注射到发育中的非洲爪蟾胚胎中，由普遍存在的和组织特异性启动子驱动，实现转基因的高效稳定表达。该策略涉及用来自腺相关病毒的反向末端重复序列（ITR）侧翼连接感兴趣的表达盒。我们的结果表明，ITR策略可能普遍适用于其他系统，如斑马鱼和胚胎干细胞，并可能在有问题的情况下实现转基因的组织特异性表达。
