Focus on genome editing and circRNA fields for many years, IntegrateRNA provides a comprehensive solution for specific knockdown of circRNA without changes of expression level of their linear counterparts. Our one-stop solution including project consulting, sgRNA design, sgRNA construction, lentivirus/AAV packaging, cell transfection, circRNA expression evaluation and cell line generation. Besides, IntergrateRNA develops several integrated vector contains RfxCas13d, crRNA and GFP, to facilitate your circRNA functional research.
Circular RNAs (circRNAs) produced from back-spliced exons are widely expressed, but individual circRNA functions remain poorly understood due to inadequate methods, such as RNAi and genome engineering, in distinguishing overlapped exons in circRNAs from those in linear cognate mRNAs. Recent studies have established that RNA-targeting type VI CRISPR effectors, known as Cas13a, Cas13b and Cas13d RNases, can be directed to cleave single-stranded RNA targets carrying complementary protospacers by a CRISPR RNA in mammalian cells. It’s reported that RfxCas13d/BSJ-gRNA system can be applied in circRNA knockdown with higher specificity and lower off-target effects. Most importantly, RfxCas13d can accurately discriminates circRNAs from linear mRNAs. Thus, RfxCas13d/BSJ-gRNA system is a simple and scalable tool that can be used to identify functional circRNAs.
Workflow of circRNA Knockdown
- sgRNA design, synthesis and construction
- Virus Packaging
- Cell transfection
- Expression evaluation of circRNA and its host gene
- Stable cell line generation
Benefits
- Guaranteed circRNA knockdown
- Lower off-target effects and lower cell toxicity
- Affordable genome-scale circRNA screening tool
- Active in vivo and in vitro, enabling the analysis RNA function in a wide range of model systems