Applications of Zinc-finger Nuclease (ZFN)

ZFN is a kind of artificially synthesized restriction endonuclease, which consists of a nonspecific cleavage domain of the Flavobacterium okeanokoites-derived FokI and a DNA-binding zinc-finger domain. At present, in many species such as plants, fruit flies, zebrafish, frogs, moose, and cattle, ZFN technology has been widely used to locate gene variations and new species with genetic backgrounds can be generated through artificial genome information. It is also of great value in the medical field, has potential significance for gene therapy of diseases, and has very broad application prospects.

Fig.1 ZFN mechanism. (Hillary & Ceasar, 2021)

ZFN-based Glycogene Editing Technology at CD BioGlyco

Gene editing is an emerging genetic engineering technology that more accurately modifies specific target genes in the genome of an organism. The gene editing tool is a sequence-specific endonuclease composed of a sequence-specific DNA binding domain and a non-specific DNA modification domain. The gene editing process is summarized as one search, two cuts, and three repairs. Identify DNA target sites on chromosomes (find), cut and generate DNA double-strand breaks (shear), and induce DNA damage repair (repair), thereby achieving directional editing of the specified genome. Simply put, gene editing is to use of a modified protein as a tool to carry out targeted modification of a specified gene.

• ZFN-mediated knockout of a glycogene
• ZFN-mediated knock-in of a glycogene
• Xenotransplantation
• Analysis
  
  • Reverse transcription polymerase chain reaction (RT-PCR)
  • Western blotting
  • Cells culture and proliferation assays, and SA-β-galactosidase assay
  • Non-homologous end joining (NHEJ) assay
  • Homologous recombination (HR) assay
  • Immunofluorescence and fluorescence-activated cell-sorting analysis
  • Mass spectrometry
  • RNA transcriptomic analysis
  • Bioinformatic analysis
• Targeted knock-in and Delivery Gene Editing

We utilize ZFNs to create complementary DNA overhangs (sticky ends) in both the genomic target site and the donor DNA allowing its integration.

Fig.2 ZFN-based glycogene editing technology. (CD BioGlyco)

Applications

• Apply for disease modeling and gene therapy development.
• To understand the etiology behind various diseases and to clarify molecular mechanisms.
• Apply to modify many genes in human cells and several model organisms.
• Development of isogenic cell systems that enable integrative polyatomic studies.
• Apply to develop the product of therapeutic glycoproteins in mammalian cells.

Advantages

• Precision technologies for genome editing bring an entirely new tool to glycobiology where it is possible to approach the biological functions of carbohydrates by gene editing screens.
• Advanced technology platform and fully custom-designed one-stop service.
• Design gene engineering events with high precision and speed.

CD BioGlyco possesses a professional platform for in vitro and in vivo Genome Editing Experiments, providing a comprehensive glycogene editing service. From study design to product/service selection, sample collection, report, and biological interpretation, our experts guide you in every step. Your research success is our goal. If you have any special requirements, welcome to feel free to contact us for custom service. We look forward to your happy cooperation.

Reference:

1. Hillary, V.E.; Ceasar, S.A. Genome engineering in insects for the control of vector borne diseases. Progress in Molecular Biology and Translational Science. 2021, 179: 197-223.