Giersch Rachael M, Finnigan Gregory C
Department of Biochemistry & Molecular Biophysics, Kansas State University, Manhattan, KS.
Yale J Biol Med. 2017 Dec 19;90(4):643-651. eCollection 2017 Dec.
The recent discovery and use of CRISPR/Cas9 gene editing technology has provided new opportunities for scientific research in many fields of study including agriculture, genetic disorders, human disease, biotechnology, and basic biological research. The ability to precisely target DNA sequences and either remove, modify, or replace genetic sequences provides a new level of control in nearly all eukaryotic organisms, including budding yeast. Given the many discoveries made in over the past decades spanning genetics, cell biology, and biochemistry, as well as the development of new technologies that have allowed high throughput screening, robotic automation, and a platform for synthetic genome engineering, the yeast community has also started to recognize the utility and complementary nature of CRISPR-based methodologies. Here we present and review a variety of recent uses of Cas9 in budding yeast-both nuclease dependent and independent applications spanning traditional gene editing and replacement, to transcriptional modulation, to novel uses including the development of living circuitry or robotic platforms for synthetic genome construction. Yeast continues to serve as a powerful model system, yet it can still benefit from use of CRISPR for basic research, industrial application, and innovation of new Cas9-based applications.
CRISPR/Cas9基因编辑技术的最新发现和应用，为包括农业、遗传疾病、人类疾病、生物技术和基础生物学研究在内的许多研究领域的科学研究提供了新机遇。精确靶向DNA序列并去除、修改或替换基因序列的能力，为几乎所有真核生物（包括芽殖酵母）提供了新的控制水平。鉴于过去几十年在遗传学、细胞生物学和生物化学方面取得的众多发现，以及高通量筛选、机器人自动化和合成基因组工程平台等新技术的发展，酵母学界也开始认识到基于CRISPR方法的实用性和互补性。在此，我们展示并回顾Cas9在芽殖酵母中的各种最新应用——从依赖核酸酶和不依赖核酸酶的应用，涵盖传统基因编辑和替换、转录调控，到包括合成基因组构建的活电路或机器人平台开发等新应用。酵母仍然是一个强大的模型系统，但它仍可从CRISPR在基础研究、工业应用和新型基于Cas9应用的创新方面的应用中受益。
