The CRISPR-Cas9 system is one of the most popular types of gene editing technology, and has been used in widespread backgrounds over the last few years. Due to its ease of use and strong extensibility, CRISPR-Cas9 has been applied to fields including laboratory research, biomedicinal research, and genetic engineering of flora and fauna. This technology can also be applied to the construction of genetically modified animal models, construction of gene-regulated cell lines, breeding of animals and plants, repair of hereditary disease, cancer research, and treatments of other serious diseases/screening of relevant drug targets.
Synbio Technologies offers sgRNA plasmid construction services for several different animal and plant species. We guarantee fast and high-quality sgRNA plasmid construction, tailored to meet a huge variety of requests and specifications.
- All model species sgRNA design : We offer sgRNA design for >20 different species.
- Many plasmid options: We can provide several validated plasmids for different species to help you best fulfill the requirements of your experiment.
- High success rate: Several target sites can be designed for different genes which enhance the success rate of plasmid construction.
- Plasmid validation service: We also provide sgRNA activity validation services, which can enhance the success rate of experiments.
1. Confirm Species and Vector
Different species and different vectors.
2. Target Site Design
We usually design 3-5 target sites on common exon for different transcription products. The important domain should be damaged as far as possible which causes loss-of-function in the target gene.
Should check the target site for single nucleotide polymorphisms (SNPs) before synthesis (If there are SNPs, T7E1 enzyme cannot be used to detect mutations).
4. Target Cutting Activity Detection
SSA activity assays and in vitro cleavage activity assay.
5. Endogenous Active Validation (Conditional Choice)
If the target cell has a high transfection rate, like 293T cell, genome can be extracted after 72h. Afterwards, amplify target sequence, verify T7E1 enzyme digestion, and validate by sequencing.