Mayo Clinic, Guggenheim, Rochester, MN 55905, USA.
Brief Funct Genomics. 2011 Jul;10(4):181-8. doi: 10.1093/bfgp/elr021. Epub 2011 Jul 10.
Morpholino oligonucleotides (MOs) are an effective, gene-specific antisense knockdown technology used in many model systems. Here we describe the application of MOs in zebrafish (Danio rerio) for in vivo functional characterization of gene activity. We summarize our screening experience beginning with gene target selection. We then discuss screening parameter considerations and data and database management. Finally, we emphasize the importance of off-target effect management and thorough downstream phenotypic validation. We discuss current morpholino limitations, including reduced stability when stored in aqueous solution. Advances in MO technology now provide a measure of spatiotemporal control over MO activity, presenting the opportunity for incorporating more finely tuned analyses into MO-based screening. Therefore, with careful management, MOs remain a valuable tool for discovery screening as well as individual gene knockdown analysis.
吗啉代寡核苷酸 (MOs) 是一种有效的、基因特异性的反义基因敲低技术，已被广泛应用于多种模式生物系统中。本文描述了 MO 在斑马鱼（Danio rerio）中的应用，用于基因功能的体内表型鉴定。我们总结了从基因靶标选择开始的筛选经验。接着，我们讨论了筛选参数的考虑因素以及数据和数据库管理。最后，我们强调了管理脱靶效应和进行彻底的下游表型验证的重要性。我们讨论了当前 MO 的局限性，包括在水溶液中储存时稳定性降低的问题。目前，MO 技术的进步提供了一种对 MO 活性的时空控制手段，为在 MO 为基础的筛选中纳入更精细的分析提供了机会。因此，只要管理得当，MO 仍然是发现筛选以及单个基因敲低分析的一种有价值的工具。
