Advances and opportunities in gene editing and gene regulation technology for Yarrowia lipolytica

Abstract

Yarrowia lipolytica has emerged as a biomanufacturing platform for a variety of industrial applications. It has been demonstrated to be a robust cell factory for the production of renewable chemicals and enzymes for fuel, feed, oleochemical, nutraceutical and pharmaceutical applications. Metabolic engineering of this non-conventional yeast started through conventional molecular genetic engineering tools; however, recent advances in gene/genome editing systems, such as CRISPR-Cas9, transposons, and TALENs, has greatly expanded the applications of synthetic biology, metabolic engineering and functional genomics of Y. lipolytica. In this review we summarize the work to develop these tools and their demonstrated uses in engineering Y. lipolytica, discuss important subtleties and challenges to using these tools, and give our perspective on important gaps in gene/genome editing tools in Y. lipolytica.

Keywords: CRISPR–Cas9; Functional genomics; Genetic tools; Genome editing; Metabolic engineering; Synthetic biology; Transposon; Yarrowia lipolytica.

Fig. 1

CRISPR–Cas9 innovations. a Hybrid promoters of RNA polymerase III promoters and tRNA lead to efficient sgRNA expression and processing. b Inclusion of gRNA target sites on the transgene plasmid lead to two possible site specific modes of integration—homologous recombination (HR) and HR-independent homology mediated end joining (HMEJ)