雅罗维亚
清脆的
基因组编辑
反式激活crRNA
Cas9
基因组工程
重组工程
计算生物学
质粒
合成生物学
基因
生物
遗传学
基因组
CRISPR干扰
出处
期刊:Methods in molecular biology
日期:2021-01-01
卷期号:: 111-121
被引量:5
标识
DOI:10.1007/978-1-0716-1414-3_7
摘要
The oleaginous yeast Yarrowia lipolytica has emerged as an industrially relevant chassis to produce various valuable chemicals. Metabolic engineering of Y. lipolytica relies on the availability of genetic engineering tools. Existing engineering strategies for this yeast include homologous recombination, random integration, and episomal plasmid-based gene expression. CRISPR-Cas9 based genome-editing toolbox has also been developed to facilitate multiplexed gene disruption and regulation. Alternative to Cas9, the CRISPR effector Cas12a has also been adopted to perform genome engineering in multiple species. Due to its distinctive features such as short and simple crRNA structure, the ability to process its own crRNA and T-rich PAM sequence (TTTN), Cas12a holds promising potential to be developed as an efficient genome-editing tool. In this chapter, we describe the protocol to implement multiplexed genome editing in Y. lipolytica. The delivery of AsCas12a and crRNA expression via a single plasmid was described. CRISPR-Cas12a-based genome editing could expand the genetic toolbox of Y. lipolytica, whihc is complementary to the classical Cas9-based tools.
科研通智能强力驱动
Strongly Powered by AbleSci AI