斑马鱼遗传操作进展

Genetic manipulation in zebrafish

摘要:

斑马鱼已经成为在发育生物学与疾病模型研究中应用最广泛的实验动物之一，其中多种遗传操作方法和高分辨率活体成像技术对斑马鱼的应用起了重要推动作用。斑马鱼在功能基因组学研究上有许多优势，如可以快速便捷地鉴定脊椎动物新基因的功能。本文尝试以示意图来简要总结应用于斑马鱼的遗传操作方法和基本原理，包括ENU化学诱变筛选的基本原理和隐性突变3代筛选的流程，逆转录病毒介导的插入突变的原理及其隐性突变3代筛选的流程，转座子Sleeping Beauty (SB) 和Tol2介导的基因捕获载体系统 (Gene-breaking transposon，GBT) 为基础的基因捕获突变的基本原理，Morpholino介导的基因下调和microRNA下调技术，TILLING (Targeting induced local lesions in genomes) 技术和CEL1分析的基本原理和实验流程，锌指核酸酶ZFN (Zinc finger nuclease) 与TALEN (Transcription activator-like effector nuclease)、CRISPR/Cas9定向基因失活的基本原理，斑马鱼上广泛采用的转基因技术的几种类型等。

Abstract:

The increasing number of genetic manipulation approaches and high-resolution live imaging technique applied in zebrafish have propelled the rise of this organism as a mainstream model for developmental biology and human diseases studies. Zebrafish has many advantages for functional genomics analysis, allowing for easy, cheap and fast functional characterization of novel genes in the vertebrate genome. Here we provide an overview of the principles of genetic manipulation in zebrafish, such as Ethylnitrosourea (ENU) mutagenesis, insertional mutagenesis, gene trapping mutagenesis, Morpholino mediated gene knockdown, targeting induced local lesions in genomes (TILLING), genome editing with engineered nucleases ZFN (Zinc finger nuclease), TALEN (Transcription activator-like effector nuclease) and CRISPR/Cas9 system, and transgenic methods used in zebrafish.