Hisano Yu, Ota Satoshi, Kawahara Atsuo
Laboratory for Cardiovascular Molecular Dynamics, RIKEN Quantitative Biology Center (QBiC), Furuedai 6-2-3, Suita, Osaka, 565-0874, Japan.
Dev Growth Differ. 2014 Jan;56(1):26-33. doi: 10.1111/dgd.12094. Epub 2013 Oct 14.
Zebrafish is a model vertebrate suitable for genetic analysis. Forward genetic analysis via chemical mutagenesis screening has established a variety of zebrafish mutants that are defective in various types of organogenesis, and the genes responsible for the individual mutants have been identified from genome mapping. On the other hand, reverse genetic analysis via targeted gene disruption using embryonic stem (ES) cells (e.g., knockout mouse) can uncover gene functions by investigating the phenotypic effects. However, this approach is mostly limited to mice among the vertebrate models because of the difficulty in establishing ES cells. Recently, new gene targeting technologies, such as the transcription activator-like effector nucleases (TALEN) and clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 systems, have been developed: that can directly introduce genome modifications at the targeted genomic locus. Here, we summarize these new and powerful genome editing techniques for the study of zebrafish.
斑马鱼是一种适合进行遗传分析的模式脊椎动物。通过化学诱变筛选进行的正向遗传分析已经建立了多种在各种器官发生类型中存在缺陷的斑马鱼突变体，并且已经从基因组图谱中鉴定出了导致各个突变体的基因。另一方面，通过使用胚胎干细胞（ES细胞）进行靶向基因破坏的反向遗传分析（例如基因敲除小鼠）可以通过研究表型效应来揭示基因功能。然而，由于建立ES细胞存在困难，这种方法在脊椎动物模型中大多仅限于小鼠。最近，已经开发出了新的基因靶向技术，如转录激活样效应核酸酶（TALEN）和成簇规律间隔短回文重复序列（CRISPR）/Cas9系统：这些技术可以直接在靶向基因组位点引入基因组修饰。在此，我们总结这些用于斑马鱼研究的新型强大基因组编辑技术。
