Comprehensive genome-scale CRISPR knockout screening of CHO cells
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 de...
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2025-01-01
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| Series: | Scientific Data |
| Online Access: | https://doi.org/10.1038/s41597-025-04438-6 |
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| author | Sung Wook Shin Su Hyun Kim Aghiles Gasselin Gyun Min Lee Jae Seong Lee |
| author_facet | Sung Wook Shin Su Hyun Kim Aghiles Gasselin Gyun Min Lee Jae Seong Lee |
| author_sort | Sung Wook Shin |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-596e15ca63094d468c60031739f58bc6 |
| institution | Kabale University |
| issn | 2052-4463 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Data |
| spelling | doaj-art-596e15ca63094d468c60031739f58bc62025-01-19T12:09:42ZengNature PortfolioScientific Data2052-44632025-01-011211910.1038/s41597-025-04438-6Comprehensive genome-scale CRISPR knockout screening of CHO cellsSung Wook Shin0Su Hyun Kim1Aghiles Gasselin2Gyun Min Lee3Jae Seong Lee4Department of Molecular Science and Technology, Ajou UniversityDepartment of Biological Sciences, KAISTDepartment of Biological Sciences, KAISTDepartment of Biological Sciences, KAISTDepartment of Molecular Science and Technology, Ajou UniversityAbstract 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.https://doi.org/10.1038/s41597-025-04438-6 |
| spellingShingle | Sung Wook Shin Su Hyun Kim Aghiles Gasselin Gyun Min Lee Jae Seong Lee Comprehensive genome-scale CRISPR knockout screening of CHO cells Scientific Data |
| title | Comprehensive genome-scale CRISPR knockout screening of CHO cells |
| title_full | Comprehensive genome-scale CRISPR knockout screening of CHO cells |
| title_fullStr | Comprehensive genome-scale CRISPR knockout screening of CHO cells |
| title_full_unstemmed | Comprehensive genome-scale CRISPR knockout screening of CHO cells |
| title_short | Comprehensive genome-scale CRISPR knockout screening of CHO cells |
| title_sort | comprehensive genome scale crispr knockout screening of cho cells |
| url | https://doi.org/10.1038/s41597-025-04438-6 |
| work_keys_str_mv | AT sungwookshin comprehensivegenomescalecrisprknockoutscreeningofchocells AT suhyunkim comprehensivegenomescalecrisprknockoutscreeningofchocells AT aghilesgasselin comprehensivegenomescalecrisprknockoutscreeningofchocells AT gyunminlee comprehensivegenomescalecrisprknockoutscreeningofchocells AT jaeseonglee comprehensivegenomescalecrisprknockoutscreeningofchocells |