Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR/Cas)

The latest addition to the tools for precision genome editing is the RNA-guided CRISPR/Cas system, an adaptive natural defense mechanism presented in bacteria and archaea against invading genetic elements. The type II CRISPR/Cas mechanism is a three-component system in which a short CRISPR RNA (crRNA), complexed with a transactivating crRNA (tracrRNA) and a Cas9 nuclease, hybridizes to a 20 bp target sequence, resulting in cleavage that destroys foreign DNA. Research has demonstrated that the Cas9 enzyme, together with a dual-tracrRNA: crRNA RNA chimera, named single-guided RNA (sgRNA), can generate site-specific double-strand break (DSB) in vitro. Another advantage of the CRISPR/Cas system is that it can be used for multiplexing by simultaneous delivery of multiple sgRNAs targeting different loci, which provides the possibility for genome-wide screenings.

Fig.1 CRISPR/Cas genome editors. (Lim & Kim, 2022)

CRISPR/Cas-based Glycogene Editing Technology at CD BioGlyco

According to the design and implementation of site-specific nuclease technologies, scientists in CD BioGlyco are committing to developing personalized therapies through classical and modern approaches. We provide transcription activator-like effector CRISPR/Cas systems services to our clients all over the world. Our services enable researchers to delete, insert, and correct any gene in a diverse range of cell types and organisms. The services we provide include but are not limited to:

• Building High-Quality CRISPR Libraries
  We are focused on building high-quality CRISPR libraries for knockout. It knocks out any gene involved in human glycosylation in arbitrary cell types.
  
  • Glycogene knockin for promoter study
  • Glycogene knockout genes at a chromosomal level
  • Glycogene activation
  • Glycogene inhibition
  • CRISPR screens
• Custom CRISPR Vectors
  We offer custom-made CRISPR vectors with high targeting efficiency and are available in multiple delivery formats (i.e. plasmid, virus, RNA). We package all major virus types to deliver your CRISPR components efficiently into difficult-to-transfect cells.
  
  • Premium-quality CRISPR plasmid packaging
  • Premium-quality CRISPR RNA packaging
  • Premium-quality CRISPR virus packaging

Fig.2 Example of CRISPR/Cas-based glycogene knockout technology. (CD BioGlyco)

Applications

• Multiplexing: CRISPR-Cas9 is often applied for multiplexing, or targeting multiple genes at once.
• Cellular delivery: CRISPR-Cas9 is delivered to cells, such as plasmid transfection or viral delivery.
• To understand the etiology behind various diseases and to clarify molecular mechanisms.

Advantages

• CRISPR-Cas9 is generally easier to design and use.
• CRISPR-Cas9 has high efficiencies in introducing breaks in the DNA, but efficiency can vary depending on the specific target sequence and the cell type.
• Simple to design and build.
• High efficiencies and target fidelity.

CD BioGlyco provides efficient and accurate Glycogene Editing Services for clients worldwide. If you have glycogene editing projects in hand, please feel free to contact us for more information.

Reference:

1. Lim, J.M.; Kim, H.H. Basic principles and clinical applications of CRISPR-based genome editing. Yonsei Med J. 2022, 63(2): 105-113.