- All-in-one dual-gRNA CRISPR-Cas9 plasmids
- Cutting-edge bioinformatics algorithm for gRNA design
- Genome editing can be conveniently confirmed by genomic PCR
Single guide RNAs are typically used when generating gene knockouts with the CRISPR-Cas9 system. This system introduces double-strand breaks, which the cell attempts to repair with non homologous end joining (NHEJ). NHEJ can be imperfect, leading to small insertions and deletions, which can cause frameshift mutations and the introduction of early stop codons. It is the introduction of these early stop codons that leads to gene knockout.
Because of the reliance on the generation of early stop codons, single guide RNA systems are not effective when knocking out noncoding RNAs, including miRNA. To address this challenge, Nawgen has developed a dual-gRNA construct system to target flanking genomic regions of the miRNA gene, leading to complete excision of the miRNA gene. This approach has many advantages:
- Complete loss of function of the miRNA gene
- Provides a larger target area to select potent gRNAs
- Large deletions can be conveniently detected and confirmed by genomic PCR instead of the more laborious T7 endonuclease assay
Additional Technical Information
Please click on the link below for more information about our dual-gRNA CRISPR system.
miRNA CRISPR Tech Note