Advancing metabolic engineering of Yarrowia lipolytica using the CRISPR/Cas system

Abstract

The oleaginous yeast Yarrowia lipolytica is widely used for the production of both bulk and fine chemicals, including organic acids, fatty acid-derived biofuels and chemicals, polyunsaturated fatty acids, single-cell proteins, terpenoids, and other valuable products. Consequently, it is becoming increasingly popular for metabolic engineering applications. Multiple gene manipulation tools including URA blast, Cre/LoxP, and transcription activator-like effector nucleases (TALENs) have been developed for metabolic engineering in Y. lipolytica. However, the low efficiency and time-consuming procedures involved in these methods hamper further research. The emergence of the CRISPR/Cas system offers a potential solution for these problems due to its high efficiency, ease of operation, and time savings, which can significantly accelerate the genomic engineering of Y. lipolytica. In this review, we summarize the research progress on the development of CRISPR/Cas systems for Y. lipolytica, including Cas9 proteins and sgRNA expression strategies, as well as gene knock-out/knock-in and repression/activation applications. Finally, the most promising and tantalizing future prospects in this area are highlighted.

Yeast-Based Biosynthesis of Natural Products From Xylose

“…Yarrowia lipolytica is an obligate aerobe that has a high flux of the TCA cycle and high translational efficiency of mitochondrial genes involved in aerobic respiration ( Man and Pilpel, 2007 ; Christen and Sauer, 2011 ; Zhu and Jackson, 2015 ; Abdel-Mawgoud et al, 2018 ; Shi et al, 2018 ; Ma et al, 2019 ). This yeast grows at temperatures below 34°C and over a wide pH range, with metabolic performances varying with cultivation conditions ( Egermeier et al, 2017 ; Abdel-Mawgoud et al, 2018 ).…”

The renaissance of yeasts as microbial factories in the modern age of biomanufacturing

“…The discovery of the CRISPR‐Cas9 genome editing system has revolutionized synthetic biology by allowing fast genetic manipulation in a variety of applications. CRISPR‐Cas9 has been implemented in a variety of S. cerevisiae backgrounds, including strains used in the brewing and wine industries (Denby et al, ; Gorter de Vries, de Groot, van den Broek, & Daran, ; Vigentini, Gebbia, Belotti, Foschino, & Roth, ) and other industrially relevant yeast species such as fission yeasts (Jacobs, Ciccaglione, Tournier, & Zaratiegui, ), Kluyveromyces marxianus (Lee et al, ), Yarrowia lipolityca (Schwartz & Wheeldon, ; Shi, Huang, Kerkhoven, & Ji, ), Ogatae species (Juergens et al, ), and Pichia species (Weninger et al, ). Of particular relevance is the development of optimized CRISPR‐Cas9 methods to engineer diploid and polyploid industrial yeast strains (Stovicek, Borja, Forster, & Borodina, ; Zhang et al, ).…”

Sustainable Animal Feed Protein through the Cultivation of YARROWIA Lipolytica on Agro-Industrial Wastes and by-Products

“…In the last five years (2015-2019), many review articles summarizing the main outcomes of this great number of studies have been released. Most reviews focus on the recent advancements concerning the cellular and metabolic engineering of the microorganism either to produce non-native high-value products [17,20,[28][29][30][31][32] or to increase/improve the productivity and yield of conventional products (e.g., citric acid, bio-lipids etc.) [13,19,[33][34][35][36][37][38].…”