Knockdown

Gene knockdown has become a commonly used approach to perform experiments in cell cultures and more recently transgenic animals. It has developed into a routine method to assess gene function in glycoengineering in a fast and easy manner. Powered by our advanced experimental technology platform and professional technical personnel, Creative Biolabs offers you a comprehensive set of gene knockdown services in cell line glycoengineering and give our global clients a guideline for glycoengineering project progression.

Background of Gene Knockdown

Different genetic techniques are in operation to investigate the functioning of genes within the living biological systems. Gene knockdown is an experimental procedure where this procedure reduces or suppresses the expression of a particular gene or genes of an organism. RNA interference (RNAi) has become a widely used approach to perform gene knockdown experiments in cell cultures and more recently transgenic animals. Gene silencing is the common definition for gene knockdown. Two types of small ribonucleic acid (sRNA) molecules, small interfering RNA (siRNA) and micro-RNA (miRNA) are central to RNAi. The primary use of this strategy is to investigate the role of the particular gene in the biological system.

Fig.1 Approaches to RNAi-mediated gene knockdown in mammalian cells.

Application in Glycoengineering

Many experimental cases demonstrate the effectiveness of gene knockdown or combinational genetic manipulation in glycosylation engineering. Engineered Chinese hamster ovary (CHO) cells with GnT-IV and -V overexpression and GalT4 knockdown express a highly glycosylated human chorionic gonadotropin (hCG) hormone. Besides, double siRNA knockdown of α1,6-fucosyltransferase (FUT8) and GDP-mannose 4,6-dehydratase (GMD) in CHO cells provide a new strategy for generating fully non-fucosylated therapeutic antibodies with enhanced ADCC. Protein sialylation was enhanced in the whole cell process after knocking down Neu3 gene expression in CHO cells. Additionally, RNAi-mediated knockdown approach was used to reduce plant-specific glycosyltransferase activities. Similar experiments are also performed in other glycoprotein production systems. For example, the engineered yeast of a Saccharomyces cerevisiae strain capable of producing a glycoprotein with humanized Man5GlcNAc2 N-linked oligosaccharides by disrupting PMT1 and PMT2 has been generated.

Features of Our Knockdown Services

• RNAi technologies
• siRNA or shRNA design and synthesis service
• siRNA or shRNA vector construction service
• mRNA/ protein expression test
• Multiple engineered CHO cell lines
• Perfect after-sale service system
• Therapeutic glycosylated antibodies with a higher ADCC function
• Cost-effective and time-saving

Creative Biolabs utilizes many different RNAi technologies to achieve robust knockdown in cell line glycoengineering. Our gene knockdown platform allows high-level expression of shRNA and miRNA that will help our clients in various scientific research. The optimized and validated technology systems are designed to minimize and eliminate the trial-and-error phase and help you get answers faster with fewer errors. Our scientists are pleased to tailor the best-fit proposal to meet your specific demands. Please feel free to contact us for more details.

Related Services:

1. Overexpression
2. Knockout and Knockin by Precision Genome Editing
3. Manipulating Heterogeneity
4. Manipulating Sialylation
5. Manipulating Fucosylation
6. Manipulating Branching
7. Manipulating Glycosaminoglycan