The CRISPR System-Keeping Zebrafish Gene Targeting Fresh
The CRISPR System-Keeping Zebrafish Gene Targeting Fresh
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DOI：
10.1089/zeb.2013.9999
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发表时间：
2013-03-01
期刊：
影响因子：
2
通讯作者：
Ekker, Stephen C.
中科院分区：
文献类型：
--
作者：
Blackburn, Patrick R.;Campbell, Jarryd M.;Ekker, Stephen C.
我们在改造和编辑模式生物基因组的能力方面正进入一个新时代。锌指核酸酶（ZFNs）在20世纪90年代末开启了首个定制核酸酶靶向基因组工程的大门。然而，由于生产困难、成本高昂以及在许多应用中效果一般，ZFNs对大多数研究实验室来说仍然遥不可及。转录激活因子样效应物核酸酶（TALENs）建立在一组植物细菌病原体中发现的DNA结合系统之上，拓宽了定制核酸酶技术，在靶向灵活性和效率方面都有显著提高。也许最重要的是，TALENs是开源的且易于生产，为世界各地的斑马鱼实验室提供了经济实惠的工具，这些工具可以在内部快速、低成本地制造，且活性可靠地高。现在，一种源自真细菌和古细菌中CRISPR/Cas系统的靶向基因组工程新系统有望进一步简化这一过程。总之，这些工具将有助于克服许多限制斑马鱼基因靶向的瓶颈，为在这种模式硬骨鱼中进行先进的基因组工程应用铺平道路。
We are entering a new era in our ability to modify and edit the genomes of model organisms. Zinc finger nucleases (ZFNs) opened the door to the first custom nuclease-targeted genome engineering in the late 1990s. However, ZFNs remained out of reach for most research labs because of the difficulty of production, high costs, and modest efficacy in many applications. Transcription activator-like effector nucleases (TALENs) were built upon a DNA binding system discovered in a group of plant bacterial pathogens and broadened custom nuclease technology, showing significant improvements in both targeting flexibility and efficiency. Perhaps most importantly, TALENs are open source and easy to produce, providing zebrafish laboratories around the world with affordable tools that can be made in-house rapidly, at low cost, and with reliably high activity. Now a new system for targeted genome engineering derived from the CRISPR/Cas system in eubacteria and archaea promises to simplify this process further. Together, these tools will help overcome many of the bottlenecks that have constrained gene targeting in zebrafish, paving the way for advanced genome engineering applications in this model teleost.