ArticleFrontiers in plant science2016
Gene Inactivation by CRISPR-Cas9 in Nicotiana tabacum BY-2 Suspension Cells.
Article in Frontiers in plant science, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 23 papers.
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Who cites it
23 citing papers in PubMed.
- CRISPR/Cas9 ribonucleoprotein mediated DNA-free genome editing in larch.Forestry research · 2024Article
- Sulfadiazine and phosphinothricin selection systems optimised for the transformation of tobacco BY-2 cells.Plant cell reports · 2023Article
- Genome editing technologies, mechanisms and improved production of therapeutic phytochemicals: Opportunities and prospects.Biotechnology and bioengineering · 2023Review
- Review
- Efficient CRISPR/Cas9-mediated genome editing in Rehmannia glutinosa.Plant cell reports · 2021Article
- Genome editing with CRISPR/Cas9 in Pinus radiata (D. Don).BMC plant biology · 2021Article
- Cellular engineering of plant cells for improved therapeutic protein production.Plant cell reports · 2021Review
- Lipofection-mediated genome editing using DNA-free delivery of the Cas9/gRNA ribonucleoprotein into plant cells.Plant cell reports · 2020Article
- New Hope for a "Cursed" Crop? Understanding Stakeholder Attitudes to Plant Molecular Farming With Modified Tobacco in Europe.Frontiers in plant science · 2020Article
- CRISPR/Cas9-mediated gfp gene inactivation in Arabidopsis suspension cells.Molecular biology reports · 2019Article
- 3D-printable portable open-source platform for low-cost lens-less holographic cellular imaging.Scientific reports · 2019Article
- Exploring the N-Glycosylation Profile of Glycoprotein B from Human Cytomegalovirus Expressed in CHO andInternational journal of molecular sciences · 2019Article
- CRISPR/Cas9-mediated homologous recombination in tobacco.Plant cell reports · 2019Article
- Application of protoplast technology to CRISPR/Cas9 mutagenesis: from single-cell mutation detection to mutant plant regeneration.Plant biotechnology journal · 2018Article
- Efficient CRISPR/Cas9-based genome editing in carrot cells.Plant cell reports · 2018Article
- Establishment of a tobacco BY2 cell line devoid of plant-specific xylose and fucose as a platform for the production of biotherapeutic proteins.Plant biotechnology journal · 2017Article
- N-Glycosylation of an IgG antibody secreted by Nicotiana tabacum BY-2 cells can be modulated through co-expression of human β-1,4-galactosyltransferase.Transgenic research · 2017Article
- Application of CRISPR/Cas9 in plant biology.Acta pharmaceutica Sinica. B · 2017Review
- Improved tissue culture conditions for the emerging CBMC biotechnology · 2017Article
- Plant genome editing with TALEN and CRISPR.Cell & bioscience · 2017Review
Corrections and comments
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Authors and funding
5 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Plant suspension cells are interesting hosts for the heterologous production of pharmacological proteins such as antibodies. They have the advantage to facilitate the containment and the application of good manufacturing practices. Furthermore, antibodies can be secreted to the extracellular medium, which makes the purification steps much simpler. However, improvements are still to be made regarding the quality and the production yield. For instance, the inactivation of proteases and the humanization of glycosylation are both important targets which require either gene silencing or gene inactivation. To this purpose, CRISPR-Cas9 is a very promising technique which has been used recently in a series of plant species, but not yet in plant suspension cells. Here, we sought to use the CRISPR-Cas9 system for gene inactivation in Nicotiana tabacum BY-2 suspension cells. We transformed a transgenic line expressing a red fluorescent protein (mCherry) with a binary vector containing genes coding for Cas9 and three guide RNAs targeting mCherry restriction sites, as well as a bialaphos-resistant (bar) gene for selection. To demonstrate gene inactivation in the transgenic lines, the mCherry gene was PCR-amplified and analyzed by electrophoresis. Seven out of 20 transformants displayed a shortened fragment, indicating that a deletion occurred between two target sites. We also analyzed the transformants by restriction fragment length polymorphism and observed that the three targeted restriction sites were hit. DNA sequencing of the PCR fragments confirmed either deletion between two target sites or single nucleotide deletion. We therefore conclude that CRISPR-Cas9 can be used in N. tabacum BY2 cells.
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