ArticleBiotechnology and bioengineering2017
MAR-Mediated transgene integration into permissive chromatin and increased expression by recombination pathway engineering.
Article in Biotechnology and bioengineering, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 15 papers.
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Who cites it
15 citing papers in PubMed, 26 citations in OpenAlex.
- Chromatin attachment to the nuclear matrix represses hypocotyl elongation in Arabidopsis thaliana.Nature communications · 2024Article
- High-efficiency and multilocus targeted integration in CHO cells using CRISPR-mediated donor nicking and DNA repair inhibitors.Biotechnology and bioengineering · 2023Article
- Current genetic strategies to investigate gene functions in Trichoderma reesei.Microbial cell factories · 2023Review
- Cell Line Development Using Targeted Gene Integration into MAR-Rich Landing Pads for Stable Expression of Transgenes.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Generation of a host cell line containing a MAR-rich landing pad for site-specific integration and expression of transgenes.Biotechnology progress · 2022Article
- Structural analysis of random transgene integration in CHO manufacturing cell lines by targeted sequencing.Biotechnology and bioengineering · 2022Article
- The high toxicity of DSB-clusters modelling high-LET-DNA damage derives from inhibition of c-NHEJ and promotion of alt-EJ and SSA despite increases in HR.Frontiers in cell and developmental biology · 2022Article
- Review
- Characterization and mutagenesis of Chinese hamster ovary cells endogenous retroviruses to inactivate viral particle release.Biotechnology and bioengineering · 2020Article
- DNA barcoding reveals that injected transgenes are predominantly processed by homologous recombination in mouse zygote.Nucleic acids research · 2020Article
- Efficient episomal gene transfer to human hepatic cells using the pFAR4-S/MAR vector.Molecular biology reports · 2019Article
- Site-specific Integration Ushers in a New Era of Precise CHO Cell Line Engineering.Current opinion in chemical engineering · 2018Article
- Autologous Cell Therapy Approach for Duchenne Muscular Dystrophy using PiggyBac Transposons and Mesoangioblasts.Molecular therapy : the journal of the American Society of Gene Therapy · 2018Article
- Unexpected phenotypic effects of a transgene integration causing a knockout of the endogenous Contactin-5 gene in mice.Transgenic research · 2018Article
- A chimeric HS4 insulator-scaffold attachment region enhances transgene expression in transfected Chinese hamster ovary cells.FEBS open bio · 2017Article
Corrections and comments
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Authors and funding
12 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Untargeted plasmid integration into mammalian cell genomes remains a poorly understood and inefficient process. The formation of plasmid concatemers and their genomic integration has been ascribed either to non-homologous end-joining (NHEJ) or homologous recombination (HR) DNA repair pathways. However, a direct involvement of these pathways has remained unclear. Here, we show that the silencing of many HR factors enhanced plasmid concatemer formation and stable expression of the gene of interest in Chinese hamster ovary (CHO) cells, while the inhibition of NHEJ had no effect. However, genomic integration was decreased by the silencing of specific HR components, such as Rad51, and DNA synthesis-dependent microhomology-mediated end-joining (SD-MMEJ) activities. Genome-wide analysis of the integration loci and junction sequences validated the prevalent use of the SD-MMEJ pathway for transgene integration close to cellular genes, an effect shared with matrix attachment region (MAR) DNA elements that stimulate plasmid integration and expression. Overall, we conclude that SD-MMEJ is the main mechanism driving the illegitimate genomic integration of foreign DNA in CHO cells, and we provide a recombination engineering approach that increases transgene integration and recombinant protein expression in these cells. Biotechnol. Bioeng. 2017;114: 384-396. © 2016 The Authors. Biotechnology and Bioengineering published by Wiley Periodicals, Inc.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.