H2-Ke6 CRISPR guide RNA, H2-K region expressed gene 6 CRISPR guide RNA[house mouse]

CRISPR gRNA for genome editing with WT SpCas9 vector or cas9 protein

The following gRNA sequences were designed by Feng Zhang’s laboratory at the Broad institute* to uniquely target the H2-Ke6 gene within the house mouse genome. These gRNA sequences are for use with WT SpCas9, or as crRNA for use with WT SpCas9 protein, to introduce a DSB for genome editing. These sgRNA sequences were validated in Sanjana N.E., Shalem O., Zhang F. Improved vectors and genome-wide libraries for CRISPR screening. Nat Methods. 2014 Aug;11(8):783-4.

• Description of H2-Ke6 CRISPR guide RNA

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  The H2-Ke6 CRISPR guide RNA sequences shown above were designed by the laboratory of Feng Zhang at the Broad Institute* in order to efficiently target the H2-Ke6 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.

• Price & Turnaround time of H2-Ke6 CRISPR guide RNA
  • $199.00/clone includes:
    • gRNA design services: Select from gRNAs above, which were designed by the Feng Zhang lab at the Broad Institute to target human and mouse genes with high specificity. Alternatively, our expert scientists will design guide RNA sequences according to your needs, for any species or genome editing strategy, when you use our online request form. We strongly recommend ordering at least 2 or more gRNAs for each target sequence.
    • gRNA synthesis and cloning into any vector of your choice, including free all-in-one vectors. The cost for gRNA constructs is $199.00/clone no matter which vector you select. Genscript offers several options for lentiviral or nonviral delivery, all suitable for use in any mammalian system. See our gRNA vector guide for details. If you prefer a different vector, simply mail an aliquot to us and we will perform subcloning at no additional cost.
  • GenCRISPR™ gRNA constructs are custom-synthesized and cloned into your choice of vector within 10 business days.
  • Our Ph.D.–level gene service representatives can answer any questions you have and help tailor our services to your needs. When you request a quote, we will contact you within 24 hours.

Our online SAM gRNA database is only available for human genes. We offer complimentary design of SAM guideRNA for mouse genes; please send email to gene@genscript.com and we will reply within 24 hours.

• Description of H2-Ke6 SAM guide RNA

  The H2-Ke6 SAM guide RNA sequences shown above will robustly activate transcription of the endogenous H2-Ke6 gene within the house mouse genome when delivered in conjunction with all required components of the SAM complex. When you order H2-Ke6 SAM gRNA constructs from GenScript, we deliver sequence-verified plasmid DNA containing your selected SAM gRNA sequences cloned into the pLenti_sgRNA(MS2)_zeo vector. You may choose to order at the same time the two other SAM plasmids: lenti dCas9-VP64_Blast and lenti MS2-P65-HSF1_Hygro.

  For complete details on the criteria and process for SAM guide RNA design and validation, please see: Konermann et al. Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature, doi:10.1038/nature14136.

• Price & Turnaround time of H2-Ke6 SAM guide RNA
  • $199.00 for each SAM gRNA sequence synthesized and cloned into the pLenti_sgRNA(MS2)_zeo vector
  • $50.00 each for lenti dCas9-VP64_Blast and lenti MS2-P65-HSF1_Hygro, which can be ordered at the same time.
  • Total price for the three-plasmid system: $299.00; additional gRNA are $199.00 each
  • 10-day turnaround time

  Our Ph.D.–level gene service representatives can answer any questions you have and help tailor our services to your needs. When you request a quote, we will contact you within 24 hours.