Related Experiment Video
Updated: Jun 15, 2026

Pooled CRISPR-Based Genetic Screens in Mammalian Cells
Published on: September 4, 2019
Comprehensive genome-scale CRISPR knockout screening of CHO cells
Sung Wook Shin1, Su Hyun Kim2, Aghiles Gasselin2
1Department of Molecular Science and Technology, Ajou University, Suwon, 16499, Republic of Korea.
Abstract:
Chinese hamster ovary (CHO) cells play a pivotal role in the production of recombinant therapeutics. In the present study, we conducted a genome-scale pooled CRISPR knockout (KO) screening using a virus-free, recombinase-mediated cassette exchange-based platform in CHO-K1 host and CHO-K1 derived recombinant cells. Genome-wide guide RNA (gRNA) amplicon sequencing data were generated from cell libraries, as well as short- and long-term KO libraries, and validated through phenotypic assessment and gRNA read count distribution. Additionally, we obtained gRNA amplicon sequencing data from the highly productive recombinant cell populations. By analyzing these datasets, essential genes involved in cell fitness as well as functional target genes associated with therapeutic protein production can be identified. Collectively, our next-generation sequencing datasets, derived from a robust and reliable CRISPR screening method, provide valuable insights into CHO genomic functions, advancing the development of next-generation CHO factories.
Insights
This study used CRISPR knockout screening in Chinese hamster ovary (CHO) cells to identify genes critical for cell fitness and therapeutic protein production, advancing CHO cell factory development.
Area of Science:
- Biotechnology
- Cell Biology
- Genomics
Background:
- Chinese hamster ovary (CHO) cells are crucial for producing recombinant therapeutics.
- Improving CHO cell productivity is essential for biopharmaceutical manufacturing.
Purpose of the Study:
- To perform a genome-scale CRISPR knockout screen in CHO-K1 cells.
- To identify genes influencing cell fitness and therapeutic protein production.
Main Methods:
- Utilized a virus-free, recombinase-mediated cassette exchange platform for CRISPR screening.
- Generated genome-wide guide RNA (gRNA) amplicon sequencing data from KO libraries.
- Validated findings through phenotypic assessment and gRNA read count analysis.
Main Results:
- Identified essential genes impacting CHO cell fitness.
- Discovered functional target genes associated with enhanced therapeutic protein production.
- Generated valuable next-generation sequencing datasets for CHO genomic function analysis.
Conclusions:
- Developed a robust CRISPR screening method for CHO cells.
- Provided insights into CHO cell biology for improved biomanufacturing.
- Advanced the development of next-generation CHO cell factories.
Related Concept Videos
CRISPR
Genetic Screens
Forward genetic screens
Forward or “classical” genetic screens involve creating random mutations in an organism’s DNA using radiation, mutagens, or insertion of additional bases, which result in visible changes...
CRISPR
CRISPR/Cas9 Genome Editing

