ArticleACS synthetic biology2018
Revealing Key Determinants of Clonal Variation in Transgene Expression in Recombinant CHO Cells Using Targeted Genome Editing.
Article in ACS synthetic biology, 2018. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
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Who cites it
19 citing papers in PubMed.
- MiRNA Chaining for Efficient Stable Overexpression to Improve Protein Quantity and Quality in CHO Cells.Methods in molecular biology (Clifton, N.J.) · 2025Article
- Targeted UDP-glucose ceramide glucosyltransferase stable overexpression induces a metabolic switch improving cell performance at high cell density.Frontiers in bioengineering and biotechnology · 2025Article
- Context-dependent genomic locus effects on antibody production in recombinant Chinese hamster ovary cells generated through random integration.Computational and structural biotechnology journal · 2024Article
- Single-Batch Expression of an Experimental Recombinant Snakebite Antivenom Based on an Oligoclonal Mixture of Human Monoclonal Antibodies.Biotechnology journal · 2024Article
- Generating Cytokines and Growth Factors for Functional Activity: Feasibility of Method Using MIF Protein.Methods and protocols · 2024Article
- Stable overexpression of native and artificial miRNAs for the production of differentially fucosylated antibodies in CHO cells.Engineering in life sciences · 2024Article
- Optimizing effector functions of monoclonal antibodies via tailored N-glycan engineering using a dual landing pad CHO targeted integration platform.Scientific reports · 2023Article
- Precise programming of multigene expression stoichiometry in mammalian cells by a modular and programmable transcriptional system.Nature communications · 2023Article
- A synthetic transcription platform for programmable gene expression in mammalian cells.Nature communications · 2022Article
- Vector design for enhancing expression level and assembly of knob-into-hole based FabscFv-Fc bispecific antibodies in CHO cells.Antibody therapeutics · 2022Article
- Improved Titer in Late-Stage Mammalian Cell Culture Manufacturing by Re-Cloning.Bioengineering (Basel, Switzerland) · 2022Article
- Article
- Hybrid cell line development system utilizing site-specific integration and methotrexate-mediated gene amplification in Chinese hamster ovary cells.Frontiers in bioengineering and biotechnology · 2022Article
- Enhanced Transgene Expression by Optimization of Poly A in Transfected CHO Cells.Frontiers in bioengineering and biotechnology · 2022Article
- Enhancement of Transgene Expression by Mild Hypothermia Is Promoter Dependent in HEK293 Cells.Life (Basel, Switzerland) · 2021Article
- PTSelect™: A post-transcriptional technology that enables rapid establishment of stable CHO cell lines and surveillance of clonal variation.Journal of biotechnology · 2021Article
- Expression vector cassette engineering for recombinant therapeutic production in mammalian cell systems.Applied microbiology and biotechnology · 2020Review
- Analytical and Functional Similarity Assessment of ABP 710, a Biosimilar to Infliximab Reference Product.Pharmaceutical research · 2020Article
- Characterization and mutagenesis of Chinese hamster ovary cells endogenous retroviruses to inactivate viral particle release.Biotechnology and bioengineering · 2020Article
Corrections and comments
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Authors and funding
8 authors.
Funding
Abstract
Generation of recombinant Chinese hamster ovary (rCHO) cell lines is critical for the production of therapeutic proteins. However, the high degree of phenotypic heterogeneity among generated clones, referred to as clonal variation, makes the rCHO cell line development process inefficient and unpredictable. Here, we investigated the major genomic causes of clonal variation. We found the following: (1) consistent with previous studies, a strong variation in rCHO clones in response to hypothermia (33 vs 37 °C) after random transgene integration; (2) altered DNA sequence of randomly integrated cassettes, which occurred during the integration process, affecting the transgene expression level in response to hypothermia; (3) contrary to random integration, targeted integration of the same expression cassette, without any DNA alteration, into three identified integration sites showed the similar response of transgene expression in response to hypothermia, irrespective of integration site; (4) switching the promoter from CMV to EF1α eliminated the hypothermia response; and (5) deleting the enhancer part of the CMV promoter altered the hypothermia response. Thus, we have revealed the effects of integration methods and cassette design on transgene expression levels, implying that rCHO cell line generation can be standardized through detailed genomic understanding. Further elucidation of such understanding is likely to have a broad impact on diverse fields that use transgene integration, from gene therapy to generation of production cell lines.
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