sgRNA Design and Validation Service

CRISPR-Cas9-based gene editing technology is a powerful new tool in biology. By simply altering the sequence of the single-guide RNA (sgRNA), one can reprogram Cas9 to target different sites in the genome with relative ease. Creative Biogene provides specialized and comprehensive sgRNA design and validation solutions for our clients' CRISPR/Cas9 experiments. Our scientists combine their extensive expertise with advanced industry technologies and are committed to creating optimized sgRNA sequences for our clients to meet their research demands.

Introduction of sgRNA Design

The performance of CRISPR/Cas relies on well-designed single-guide RNA (sgRNA). Since sgRNAs are responsible for recruiting Cas9 to specific genomic sites for targeted cleavage, the design of sgRNAs is critical to the success of gene editing experiments. The sgRNA is derived from the fusion of natural tracrRNA (transactivating crRNA) and crRNA (CRISPR RNA). Taking the widely used spCas9 system as an example, before designing sgRNAs, PAM sequences should be searched for near the target genome (PAM sequences are NGG, N=A/T/C/G), and gene-specific sgRNA template sequences are the neighboring motifs located in the sequence adjacent to the pre-interval region of the PAM sequence. The sgRNA, by directing the Cas 9 protein, can be sheared 3-4 bases upstream of the PAM sequence, inducing cells to undergo double-stranded breaks and form mutations in the target gene.

Fig. 1 CRISPR/Cas9 genome editing system. (Cui, Y., et al, 2018)

One-Stop sgRNA Design and Validation Service at Creative Biogeen

Creative Biogene provides customers with one-stop sgRNA services, including sgRNA design, sequence optimization, sgRNA construction and validation services. Our comprehensive service portfolio can provide customers with high-quality sgRNA sequences of interest in the shortest possible time to support their gene editing experiments.

Our sgRNA design service

• Custom sgRNA design
  Creative Biogene utilizes advanced bioinformatics tools to aid in the design of highly active and specific sgRNAs. Our expert team designs sgRNAs for the genes of interest to our customers, and our extensive project experience combined with advanced design tools can effectively minimize off-target effects and improve editing efficiency to ensure the success of gene editing.
• Sequence optimization
  We can analyze and optimize the design of existing sequences to maximize CRISPR-Cas9 activity and minimize off-target effects.

Our sgRNA validation service

• sgRNA validation
  After the construction of sgRNAs against a target gene or sequence is completed, activity detection experiments at the site need to be performed. Creative Biogene enables rapid detection of sgRNA activity by using validated sgRNA target screening kits as well as cell-based assays and other methods, which include:
• Rapid high-throughput activity screening and validation services using sgRNA target screening kits.
• Transfection of sgRNA-pSpCas9 into 293 cells, extraction of genomic DNA from 293 cells, PCR amplification of target DNA fragments followed by the use of T7 nucleic acid endonuclease to verify sgRNA cleavage efficiency.
• Off-target analysis
  At Creative Biogene, various experimental systems are widely used to study the off-target effects of sgRNA. Our team uses many different types of in vitro assay-based, single-molecule, computational, and crystallographic methods to detect and quantify off-target effects induced by CRISPR/Cas9 and other CRISPR/Cas systems, ensuring sgRNA specificity.

Creative Biogene ensures reliable sgRNA design and validation service with competitive pricing and fast turnaround time. Contact us to learn more about our services.