Nicotiana tabacum Engineering Service
Nicotiana tabacum Engineering Service at CD BioGlyco
Nicotiana tabacum is an annual or perennial herbaceous plant of the Solanaceae family, native to South America and now widely cultivated around the world. It has been extensively studied for bioengineering applications due to its high biomass, fast growth rate, and simple genetic manipulation. It is one of the important Plant Chassis for synthetic biology we provide. With the advanced GlycoChas™ Cells platform, CD BioGlyco provides comprehensive N. tabacum engineering service. The services we provide include but are not limited to the following.
Gene editing
Gene editing is one of the important means to engineer tobacco in synthetic glycobiology. Using gene editing systems such as CRISPR/Cas9 and CRISPR/Cpf1, we precisely edit the tobacco genome to achieve gene knockout, knock-in, or site-directed mutation.
Gene knockout: By designing a specific gRNA (guide RNA), the Cas protein can locate the target gene and cut its DNA sequence, thereby achieving the knockout of the gene. This helps to study the function of genes or eliminate negative effects on metabolic pathways.
Gene knock-in: Gene knock-in technology allows scientists to insert foreign genes or regulatory sequences at specific locations to enhance target metabolic pathways or introduce new metabolic capabilities.
Site-directed mutagenesis: Through precise gene editing, specific genes can be fine-tuned without destroying the functions of other genes, such as optimizing enzyme activity, changing substrate specificity, or improving metabolic flux.
Glycoengineering (metabolic engineering)
Glycoengineering focuses on optimizing or introducing new metabolic pathways to produce specific glycan compounds. in tobacco.
Pathway optimization: We optimize the synthesis and accumulation of glycans in tobacco by enhancing or weakening the expression of key enzymes and adjusting the direction and intensity of metabolic flow.
Introduction of heterologous pathways: We introduce metabolic pathways from other species into tobacco, allowing it to synthesize glycan compounds that are not produced by itself. This often involves the co-expression of multiple genes to ensure pathway integrity and efficiency.
Metabolic flow analysis: We use metabolomics, flux analysis, and other methods to comprehensively evaluate the dynamic changes of metabolic flow in tobacco cells and provide data support for the optimization of metabolic pathways.
Glycan analysis
Glycan Analysis technology is an integral part of synthetic glycobiology and is used to identify and quantify glycan compounds synthesized in tobacco.
Glycan extraction and purification: We develop efficient extraction and purification methods to isolate target glycosides from tobacco tissue and provide pure samples for subsequent analysis.
Structural identification: We use advanced analytical technologies such as nuclear magnetic resonance (NMR) and mass spectrometry (MS) to conduct structural identification of the extracted glycosides and determine their chemical composition and spatial configuration.
Quantitative analysis: We use chromatography, MS, and other methods to accurately measure the content of glycan compounds in tobacco and evaluate the impact of engineering modification on glycan synthesis.
Publication
Technology: Glycoprotein expression and modification
Journal: Molecular genetics and metabolism
IF: 4.204
Published: 2015
Results: The authors developed a new chemically modified α-Gal-A known as PRX-102, which was expressed in N. tabacum. The properties of PRX-102, both in vivo and in vitro, were characterized by the authors and subsequently compared with two commercially available α-galactosidase-A enzymes. The analysis revealed that after an incubation period of 1 h, PRX-102 demonstrated 30% activity, indicating its prolonged in vitro stability in the plasma and contrasting with the complete inactivation of the commercial enzymes. The authors also discovered that in male Fabry mice, PRX-102's pharmacokinetic profile demonstrated a 10-fold increase in t1/2 in comparison to the approved drugs. PRX-102 was found to be characterized by a relatively simple plant-like glycosylation pattern, typically featuring tri-mannose structures along with an additive of either an α(1–3)-linked fucose or a β(1–2)-linked xylose.
Here are some of the results shown in this article:
Fig.1 In vitro stability of PRX-102, agalsidase alfa and agalsidase beta. (Kizhner, et al., 2015)
Advantages of Us
- Possessing an efficient metabolic engineering platform: N. tabacum, as a fast-growing plant, has efficient metabolic pathways and strong biosynthetic capabilities. By utilizing these characteristics of tobacco plants, we provide clients with efficient and rapid production services, helping clients achieve large-scale synthesis of target compounds.
- Cost-effectiveness and sustainability: The use of plant engineering to produce compounds is more cost-effective and sustainable than traditional chemical synthesis methods. By harnessing the biosynthetic capabilities of tobacco plants, we achieve low-cost production of compounds and reduce negative environmental impacts.
- Professional knowledge and technical support: As a professional biotechnology company, we have rich professional knowledge and technical experience. We provide our clients with comprehensive N. tabacum engineering service.
CD BioGlyco provides clients with excellent N. tabacum engineering services including gene editing service, metabolic engineering service, and glycan analysis service. We sincerely welcome clients to contact us if you are interested in our N. tabacum engineering service.
Reference
- Kizhner, T.; et al. Characterization of a chemically modified plant cell culture expressed human α-Galactosidase-A enzyme for treatment of Fabry disease. Molecular genetics and metabolism. 2015, 114(2): 259-267.
For research use only. Not intended for any clinical use.
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