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CRISPR/Cas9 has been widely utilized to facilitate functional knockout of protein coding genes. However, a single site of gene editing may not be sufficient to disrupt protein’s structure (for example, miRNA stem-loop) and function. To overcome this potential limitation, paired guide RNAs targeting both 5’ and 3’ of the target region have been shown to result in larger deletions, thus resulting in loss-of-function completely.
Dual sgRNA-directed large gene deletion mediated by CRISPR/Cas9 system is a robust tool for generating deletions of long non-coding RNAs (lncRNAs), MicroRNAs (miRNAs) and regulatory sequences [1,2]. Several recent studies have reported the CRISPR/ Cas9 system-induced large genomic deletion, ranged from 23kb to 1Mb, in mouse , C. elegans , rabbit  and human cell lines .
Creative Biogene is experienced in producing a fragment deleted cell line using any mammalian cell line and targeting sequences of your interest. Utilization of two sgRNAs targeting the immediate sequence upstream and downstream is a viable method for functional knockout of a target protein completely.
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