Trichoplusia ni, also known as the cabbage looper, is a common pest of crops whose derivative cell line (High-Five) is frequently used in biotechnology, including synthetic glycobiology. The application of Trichoplusia ni engineering in synthetic glycobiology involves cell engineering, gene editing, recombinant expression system construction, glycosylation analysis, enzyme engineering, glycan chain synthesis, and metabolic engineering. These technologies promote the development and application of Trichoplusia ni as a synthetic glycobiology research platform.

Trichoplusia ni Engineering Service at CD BioGlyco

At CD BioGlyco, GlycoChas™ Cells is a platform containing multiple types of chassis, including Insect Chassis, which helps to provide clients with professional and efficient Trichoplusia ni engineering services.

• Cell Engineering

• Cell line establishment and culture. We select Trichoplusia ni cell lines suitable for synthetic glycobiology first. These cell lines should have efficient foreign protein expression capabilities. Then the cell growth rate and protein expression level are increased by optimizing the culture medium formula and adjusting culture conditions (such as temperature, pH value, and dissolved oxygen).

• Cell transformation. Viral vectors or non-viral vectors are used to introduce foreign genes into Trichoplusia ni cells to achieve efficient expression of specific glycosyltransferases or glycan chain synthesis-related genes.

• Gene Editing

• CRISPR-Cas9-mediated gene editing. This technology involves steps such as target design, gene knockout, and gene knock-in.

• RNA interference (RNAi). A specific RNAi sequence is designed for the target gene first. The target gene is then silenced through RNAi technology to study the impact of the gene on the glycosylation process.

• Recombinant Expression System Construction

The baculovirus expression vector system (BEVS) is a system to express recombinant proteins in insect cells such as those of Trichoplusia ni. This system uses baculovirus vectors to introduce foreign DNA into insect cells, leading to large-scale production of the desired protein.

• Glycosylation Analysis

• Glycan chain structure analysis. In general, mass spectrometry (MS) and nuclear magnetic resonance (NMR) are used to identify and resolve the precise structure of glycan chains.

• Glycosylated protein detection. Western blotting is used to detect the expression and glycosylation degree of glycosylated proteins. Visualize the localization and distribution of glycosylated proteins in cells using fluorescent labeling technology.

• Enzyme Engineering and Glycan Chain Synthesis

• Expression and purification of glycosyltransferase. Increase the expression level of glycosyltransferase by adjusting cell culture conditions and transfection parameters first. The active glycosyltransferase is then purified from the cells using techniques such as chromatography and electrophoresis.

• In vitro glycan chain synthesis. Purified glycosyltransferases are used to synthesize glycan chains of specific structures in vitro. Optimize the efficiency and product quality of in vitro glycan chain synthesis by adjusting reaction conditions and adding cofactors.

• Metabolic Engineering

• Metabolic pathway analysis. To reconstruct and optimize the glycan synthesis pathway in Trichoplusia ni by analyzing the glycosylation metabolic pathway.

• Metabolic flow regulation. To increase the rate and yield of glycan chain synthesis by increasing the supply of precursor substances, (such as glycan sources and cofactors). To reduce competition and inhibition of glycan chain synthesis by inhibiting the production of metabolic by-products.

Applications of Trichoplusia ni Engineering

• Cell lines from Trichoplusia ni can be used to express glycosyltransferases from different organisms. These enzymes catalyze transfer reactions between glycan molecules to synthesize complex glycan chain structures. Highly active glycosyltransferases can be obtained to provide enzyme sources for subsequent glycosylation reactions by optimizing expression conditions and purification methods.
• Trichoplusia ni cell lines can be used to express glycoproteins or glycolipids with specific glycosylation sites. Glycan chains with specific structures and functions can be synthesized on these molecules by regulating the intracellular glycosylation pathway and introducing exogenous glycosyltransferases.
• Trichoplusia ni engineering can be used to construct mutant libraries with different glycan chain structures to study the impact of glycan chain structures on biological activity. Mutants with optimized glycan chain structures can be screened to improve their biological activity or stability using high-throughput screening technology.
• The glycosylation pathway of the Trichoplusia ni cell line is modified through gene editing technology to introduce new glycosyltransferases or regulate the activity of existing enzymes. These modifications expand the glycosylation capabilities of the cell lines, allowing them to synthesize a wider variety of glycan structures.

Advantages of Us

• We have profound professional knowledge and technical background, and has rich experience and strength in the field of synthetic glycobiology. We provide professional services in all aspects of insect cell engineering, gene editing, protein expression, and purification.
• We provide personalized synthetic glycobiology solutions based on specific needs to meet clients' requirements.
• We are committed to continuous innovation and development, and constantly explores new technologies to improve the efficiency and quality of synthetic glycobiology. We continue to accumulate experience and optimize experimental processes to provide clients with better and more efficient services.

With professional technology, CD BioGlyco provides comprehensive Trichoplusia ni engineering service. We use advanced biotechnology and engineering methods to optimize Trichoplusia ni chassis and produce high-quality glycan and glycoprotein products. Our services cover the entire process from gene editing to product purification, ensuring clients receive high-quality glycobiology solutions. Clients interested in Trichoplusia ni engineering service are welcome to contact us.

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