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Specializing in circRNA research and genome editing technology for years, IntergrateRNA provides circRNA knockout service using CRISPR/Cas9 technology for global customers. Our comprehensive circRNA knockout service covers from project design, sgRNA selection, vector construction, cell line transfection, expression analysis of target circRNA and host gene to stable cell line generation. Our mission is to offer fully customized and high-quality service to meet your requirements.
Covalently closed circular RNAs (circRNAs) are produced by precursor mRNA back-splicing of exons of thousands of genes in eukaryotes. circRNAs are widely expressed in various organisms in a specie-, tissue-, disease- and developmental stage-specific manner, and have been demonstrated to play a vital role in the pathogenesis and progression of human diseases, including cancer, diabetes, cardiovascular disease, chronic inflammation and nervous system diseases. With the development of biotechnology and more in-depth research on circRNA, circRNA will play important roles in disease prevention, diagnosis and treatment discovery.
Although circRNA has proven to function as miRNA sponges, RBP sponges, protein translation templates and regulatory factor of transcription/splicing, the function of most circRNAs remain largely unexplored. Thus, it's necessary to assess the function of circRNAs by loss of function experiment like CRISPR/Cas9 mediated knockout.
Two sgRNAs are designed to targeting the ends of circRNA-forming exon, to achieve large fragment deletion of circRNA. But it will inevitably affect linear RNA (host gene) expression because sequences of circRNAs totally overlap with linear RNAs.
Two sgRNA are designed to targeting the intronic complement sequence (ICS) of the circRNA-flanking introns, to minimize circRNA generation without affecting the expression of residing protein-coding gene. Removal of the intronic unilateral ICS by the CRISPR/Cas9 system to disrupt the formation of RNA pairing, in principle, was able to reduce circRNA expression or, in some circumstances, to completely knock out a circRNA. It's an ideal method for circRNA knockout.
Figure 1. Strategies of circRNA knockout
Based on a team of experts and a professional platform, IntergrateRNA provides customers with high-quality and efficient services related to circRNA knockout. We offer two different strategies to completely knock out a circRNA with/without affecting the expression of the host gene. For further information of circRNA, please feel free to contact us at firstname.lastname@example.org.