PIKFYVE CRISPR guide RNA, phosphoinositide kinase, FYVE-type zinc finger containing CRISPR guide RNA[human]
gRNA/crRNA 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 PIKFYVE gene within the human 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 PIKFYVE CRISPR guide RNA
The PIKFYVE CRISPR guide RNA sequences shown above were designed by the laboratory of Feng Zhang at the Broad Institute* in order to efficiently target the PIKFYVE 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 PIKFYVE 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.
gRNA for transcription activation with SAM
The following gRNA sequences were designed by Feng Zhang’s laboratory at the Broad institute* to uniquely and robustly activate transcription of the endogenous PIKFYVE gene within the human genome when used with the the CRISPR/Cas9 Synergistic Activation Mediators (SAM) complex. These gRNA specifically target the first 200 bp upstream of the transcription start site (TSS). These validated sgRNA sequences were published in Konermann S et al. Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature, 2015 Jan 29;517(7536):583-8.
Description of PIKFYVE SAM guide RNA
The PIKFYVE SAM guide RNA sequences shown above will robustly activate transcription of the endogenous PIKFYVE gene within the human genome when delivered in conjunction with all required components of the SAM complex. When you order PIKFYVE 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 PIKFYVE 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.
Related Services of PIKFYVE CRISPR guide RNA
- GenCRISPR™ mammalian cell line service for fully-validated KO or KI cell lines.
- Gene Synthesis for knock-in plasmids.
- Codon optimization to optimize Cas9 for different host species
- Plasmid Prep to get the quantity and sterility your experiments require.
Related gene information of PIKFYVE CRISPR guide RNA
|Entre Gene ID||200576|
|Full Name||phosphoinositide kinase, FYVE-type zinc finger containing|
|Summary||Phosphorylated derivatives of phosphatidylinositol (PtdIns) regulate cytoskeletal functions, membrane trafficking, and receptor signaling by recruiting protein complexes to cell- and endosomal-membranes. Humans have multiple PtdIns proteins that differ by the degree and position of phosphorylation of the inositol ring. This gene encodes an enzyme (PIKfyve; also known as phosphatidylinositol-3-phosphate 5-kinase type III or PIPKIII) that phosphorylates the D-5 position in PtdIns and phosphatidylinositol-3-phosphate (PtdIns3P) to make PtdIns5P and PtdIns(3,5)biphosphate. The D-5 position also can be phosphorylated by type I PtdIns4P-5-kinases (PIP5Ks) that are encoded by distinct genes and preferentially phosphorylate D-4 phosphorylated PtdIns. In contrast, PIKfyve preferentially phosphorylates D-3 phosphorylated PtdIns. In addition to being a lipid kinase, PIKfyve also has protein kinase activity. PIKfyve regulates endomembrane homeostasis and plays a role in the biogenesis of endosome carrier vesicles from early endosomes. Mutations in this gene cause corneal fleck dystrophy (CFD); an autosomal dominant disorder characterized by numerous small white flecks present in all layers of the corneal stroma. Histologically, these flecks appear to be keratocytes distended with lipid and mucopolysaccharide filled intracytoplasmic vacuoles. Alternative splicing results in multiple transcript variants encoding distinct isoforms.[provided by RefSeq, May 2010].|
Related Services of PIKFYVE CRISPR guide RNA
Learn more about the CRISPR gRNA constucts.
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