The Sema5b CRISPR guide RNA sequences shown above were designed by the laboratory of Feng Zhang at the Broad Institute* in order to efficiently target the Sema5b gene with minimal risk of off-target Cas9 binding elsewhere in the genome. For complete details on the criteria and process for guide RNA design and selection, please see: Sanjana N.E., Shalem O., Zhang F. Improved vectors and genome-wide libraries for CRISPR screening. Nat Methods. 2014 Aug;11(8):783-4. doi: 10.1038/nmeth.3047. Read the Full Text
Based on our experience using our design tool to create knock-out cell lines, a single gRNA construct is typically sufficient to knock-out your gene of interest, but to increase your chance of success, we recommend ordering at least two gRNA constructs per gene that you want to target. We recommend that you double-check the gRNA sequences against your target gene sequence of interest before ordering, especially if you are trying to target only one specific splice variant or a specific exon.
When you order gRNA clones from GenScript, we deliver a sequence-verified plasmid containing all elements required for gRNA expression and genome binding: the U6 promoter, spacer (target) sequence, gRNA scaffold, and terminator. You may select from vectors that include selection markers. We guarantee sequence accuracy for gRNA clones we deliver; however, given the complexity of creating genomically edited cell lines, we cannot guarantee the outcome of experiments using our gRNA constructs. If you prefer to receive sequence-validated KO or KI cell lines created using CRISPR technology, please refer to our GenCRISPR™ mammalian cell line service.
