Principle of Glycogene Tagging

Glycogene tagging is usually used to test the success of recombinant DNA vector transformation or to test the localization of the target gene in cells or tissues. Commonly used marker genes are some antibiotic resistance genes. For example, after combining a plasmid with a resistance gene and foreign DNA fragments to form a recombinant plasmid, then is transferred into the recipient cell. It judges whether the recipient cell has acquired the target gene according to whether the recipient cell is resistant. When recipient cells are grown with selective media, recipient cells that survive the medium can be considered successful in foreign DNA. Different glycogene tags have different uses, which facilitate the development of glycogenomics. Glycogene tagging has broad applications in general genome engineering, protein production, and immune cell therapy.

Fig.1 Gene tagging strategy at pGAPDH locus. (Xiang, et al., 2019)

Glycogene Tagging Service at CD BioGlyco

The development of glycogene tagging has facilitated the expression of glycogenic diseases and cell lines, CD BioGlyco provides efficient glycogene tagging services. The methods we provide have been repeatedly validated for different targets and target genes and provide you with accurate and reliable tagging services.

• The first one is a kit that does not require the preparation of donor plasmid, and combined with efficient gene editing tools, the labeling efficiency is close to 100%.
• The second is to label the gene with fluorescent labeling of the C- or N-terminus of the glycogen or labeling of the glycopeptide with the help of clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated nuclease 9 (Cas9) system. The advantage of generating fluorescently labeled genes is that the subcellular location is easily tracked, and the gene expression and dynamics are monitored in real time under different conditions.
• The third is small molecule tagging genes, which are easily purified and detected using standardized antibodies, solving the problem of non-specific or unavailable antibodies.

Fig.2 Classification of technical approaches to glycogene tagging. (CD BioGlyco)

Applications

• Glycogene tagging plays a key role in the development of glycogenic selection microarrays.
• Glycogene tagging is used in the marking of disease resistance-related factors.
• Glycogene tagging is suitable for direct simultaneous detection of an endogenous mRNA and its translated protein product in the same cell.

Advantages of Us

• The true tag enables rapid production of donor DNA that contains an antibiotic selection marker to enrich cells containing the tagged gene.
• The tagging approach we present uses standard probe sets to generate a clear cellular picture of allelic mRNA expression.

CD BioGlyco has always adhered to the client-centric principle and developed an advanced Glycogenomics Platform. We have already had many successful analysis cases and won unanimous praise from clients worldwide. In addition, we provide precise Glycogene Knockout/Knock-in/Knockdown/Overexpression Services. Please feel free to contact us if you are interested in our services and have any questions.

Reference:

1. Xiang, X., et al. CRISPR/Cas9-mediated gene tagging: a step-by-step protocol. Methods Mol Biol. 2019, 1961: 255-269.