ArticleBiotechnology journal2018
Single Copy Transgene Integration in a Transcriptionally Active Site for Recombinant Protein Synthesis.
Article in Biotechnology journal, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 12 papers.
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The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
12 citing papers in PubMed, 45 citations in OpenAlex.
- Context-dependent genomic locus effects on antibody production in recombinant Chinese hamster ovary cells generated through random integration.Computational and structural biotechnology journal · 2024Article
- Integrating Omics and CRISPR Technology for Identification and Verification of Genomic Safe Harbor Loci in the Chicken Genome.Biological procedures online · 2023Article
- Structural analysis of random transgene integration in CHO manufacturing cell lines by targeted sequencing.Biotechnology and bioengineering · 2022Article
- The Challenges in Developing Efficient and Robust Synthetic Homing Endonuclease Gene Drives.Frontiers in bioengineering and biotechnology · 2022Review
- Enhancing stability of recombinant CHO cells by CRISPR/Cas9-mediated site-specific integration into regions with distinct histone modifications.Frontiers in bioengineering and biotechnology · 2022Article
- High-resolution three-dimensional chromatin profiling of the Chinese hamster ovary cell genome.Biotechnology and bioengineering · 2021Article
- Cell culture bioprocessing - the road taken and the path forward.Current opinion in chemical engineering · 2020Article
- Identifying Improved Sites for Heterologous Gene Integration Using ATAC-seq.ACS synthetic biology · 2020Article
- Multiplexed clonality verification of cell lines for protein biologic production.Biotechnology progress · 2020Article
- Predicting favorable landing pads for targeted integrations in Chinese hamster ovary cell lines by learning stability characteristics from random transgene integrations.Computational and structural biotechnology journal · 2020Article
- Model-Driven Engineering of N-Linked Glycosylation in Chinese Hamster Ovary Cells.ACS synthetic biology · 2019Article
- Recurring genomic structural variation leads to clonal instability and loss of productivity.Biotechnology and bioengineering · 2019Article
Corrections and comments
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Authors and funding
11 authors at 2 institutions in 1 country.
Funding
Abstract
For the biomanufacturing of protein biologics, establishing stable cell lines with high transgene transcription is critical for high productivity. Modern genome engineering tools can direct transgene insertion to a specified genomic locus and can potentially become a valuable tool for cell line generation. In this study, the authors survey transgene integration sites and their transcriptional activity to identify characteristics of desirable regions. A lentivirus containing destabilized Green Fluorescent Protein (dGFP) is used to infect Chinese hamster ovary cells at a low multiplicity of infection, and cells with high or low GFP fluorescence are isolated. RNA sequencing and Assay for Transposase Accessible Chromatin using sequencing data shows integration sites with high GFP expression are in larger regions of high transcriptional activity and accessibility, but not necessarily within highly transcribed genes. This method is used to obtain high Immunoglobulin G (IgG) expressing cell lines with a single copy of the transgene integrated into transcriptionally active and accessible genomic regions. Dual recombinase-mediated cassette exchange is then employed to swap the IgG transgene for erythropoietin or tumor necrosis factor receptor-Fc. This work thus highlights a strategy to identify desirable sites for transgene integration and to streamline the development of new product producing cell lines.
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Registered trials
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