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Purpose
The GlycoGene CRISPR lentiviral library targets human genes involved in cellular glycosylation. These include glycosyltransferase genes involved in biosynthesis of several different types of glycoconjugates, including: N-glycans, O-glycans, glycolipids, glycosaminoglycans, GPI-anchored carbohydrates, O-fucose, O-GlcNAc, O-Glucose, O-Mannose, and EGF domain specific O-linked GlcNAcTs (EOGTs).
This library also targets proteins involved in intracellular protein transport, sulfotransferases, enzymes involved in nucleotide-sugar biosynthesis, nucleotide-sugar transporters and glycosidases.
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Vector Backbone
SiC-V2-Cas9G7 - does not express Cas9
This vector was modified as described in the associated publication. The mCerulean gene has been replaced with TagBFP. For library construction the sgRNA cloning region of lentiCRISPR V2 was cloned downstream of the U6 promoter.
The full backbone vector sequence can be found in the Resource Information section.
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Depositing Labs
Ordering
| Item | Catalog # | Description | Quantity | Price (USD) | ||
|---|---|---|---|---|---|---|
| Pooled Library | 140961 | Human GlycoGene CRISPR pooled library† | 1 | $ 380 | Add to Cart | |
† A Cas9 plasmid is NOT included with this item and will have to be ordered separately. The library can be used in conjunction with any SpCas9 expressing plasmid, such as lentiCas9-Blast (Addgene #52962) or SiC-V1 (Addgene #133041). Alternatively, cell lines expressing SpCas9 or cells electroporated with Cas9 protein can be used for screening.
Library Details
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SpeciesHuman
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Genes targeted347
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gRNAs3637
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ControlsNon-targeting gRNAs
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Lentiviral Generation3rd
Library Shipping
This library is delivered as suspended DNA in a microcentrifuge tube on blue ice. The tube's contents will not necessarily be frozen. For best results, minimize freeze-thaws.
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Volume∼10 µL
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Concentration10 ng/µL
Resource Information
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Protocols
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Depositor Data
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Terms and Licenses
Depositor Comments
The depositing lab has created a web resource to facilitate use of the GlycoGene CRISPR Library: GlycoGene CRISPR Library Viewer
They recommend using MAGeCK/MAGeCK-VISPR algorithms for analysis.
These pooled libraries were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.
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For your Materials & Methods section:
Human GlycoGene CRISPR library was a gift from Sriram Neelamegham (Addgene #140961) -
For your References section:
A GlycoGene CRISPR-Cas9 lentiviral library to study lectin binding and human glycan biosynthesis pathways. Zhu Y, Groth T, Kelkar A, Zhou Y, Neelamegham S. Glycobiology. 2020 Aug 8. pii: 5889976. doi: 10.1093/glycob/cwaa074. PubMed 32776087