CRISPR-Cas9-mediated genomic multiloci integration in Pichia pastoris.

ABSTRACT:

Background

Pichia pastoris (syn. Komagataella phaffii) is a widely used generally recognized as safe host for heterologous expression of proteins in both industry and academia. Recently, it has been shown to be a potentially good chassis host for the production of high-value pharmaceuticals and chemicals. Nevertheless, limited availability of selective markers and low efficiency of homologous recombination make this process difficult and time-consuming, particularly in the case of multistep biosynthetic pathways. Therefore, it is crucial to develop an efficient and marker-free multiloci gene knock-in method in P. pastoris.

Results

A non-homologous-end-joining defective strain (Δku70) was first constructed using the CRISPR-Cas9 based gene deficiency approach. It was then used

SUBMITTER: Liu Q 

PROVIDER: S-EPMC6704636 | biostudies-literature | 2019 Aug

REPOSITORIES: biostudies-literature