Efficient site-specific integration in CHO-K1 cells using CRISPR/Cas9-modified donors.

ABSTRACT:

Background

Conventional methods applied to develop recombinant CHO (rCHO) cell line as a predominant host for mammalian protein expression are limited to random integration approaches, which can prolong the process of getting the desired clones for months. CRISPR/Cas9 could be an alternative by mediating site-specific integration into transcriptionally active hot spots, promoting homogenous clones, and shortening the clonal selection process. However, applying this approach for the rCHO cell line development depends on an acceptable integration rate and robust sites for the sustained expression.

Methods and results

In this study, we aimed at improving the rate of GFP reporter integration to the Chromosome 3 (Chr3) pseudo-attP site of the CHO-K1 genome via two strategies; the

SUBMITTER: Kheirandish MH 

PROVIDER: S-EPMC10224643 | biostudies-literature | 2023 Jul

REPOSITORIES: biostudies-literature