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A validated gRNA library for CRISPR/Cas9 targeting of the human glycosyltransferase genome.

Narimatsu Y, Joshi HJ, Zhang Y, Gomes C, Chen YH, Lorenzetti F, Furukawa S, Schjoldager K, Hansen L, Clausen H, Bennett EP, Wandall HH
Glycobiology. 2018 Jan 5. pii: 4791732. doi: 10.1093/glycob/cwx101. (Link opens in a new window) PubMed (Link opens in a new window) Article

GlycoCRISPR collection

More than 200 glycosyltransferases are involved in biosynthesis of the human glycome, which is comprised of all glycan structures found on different glycoconjugates in all cells. In short of the necessary analytic strategies that enable researchers to decipher the biological roles of any glycan with respect to structure and/or function, the biosynthetic roles of most of these enzymes have been defined. Thus, GlycoCRISPR provides the first collection of CRISPR/Cas9 gRNAs targeting all human glycosyltransferase encoded genes (glycogenes). The uniqueness of this collection becomes evident from the fact that all gRNA designs deposited in the collection have been experimentally validated by recently published methods (Lonowski et al., 2017) to give rise to >30% target cutting efficiency based on in depth indel profiling by IDAA (Yang et al., 2015). GlycoCRISPR gRNAs are made available as a sustainable plasmid format encoding U6 promoter driven gRNA expression. The promised high indel formation capacity of all glycogene gRNA designs enables easy genetic dissection of glycan biosynthetic pathways either as single and/or multiplex combinatorial targeting of the human glycosyltransferase genome.

Workflow for establishment of the GlycoCRISPR collection 205.3 KB

Additional References

  • Lonowski LA, Narimatsu Y, Riaz A, Delay CE, Yang Z, Niola F, Duda K, Ober EA, Clausen H, Wandall HH, Hansen SH, Bennett EP, Frödin M. 2017. Genome editing using FACS enrichment of nuclease-expressing cells and indel detection by amplicon analysis. Nat Protoc. Mar;12(3):581-603. PMID: 28207001
  • Yang Z, Steentoft C, Hauge C, Hansen L, Thomsen AL, Niola F, Vester-Christensen MB, Frödin M, Clausen H, Wandall HH, Bennett EP. 2015. Fast and sensitive detection of indels induced by precise gene targeting. Nucleic Acids Res. May 19;43(9):e59. PMID: 25753669

Plasmids from Article

ID Plasmid Purpose
106682gh1expression of gRNA targeting GALNT1
106683gh10expression of gRNA targeting GALNT10
106684gh11expression of gRNA targeting GALNT11
106685gh12expression of gRNA targeting GALNT12
106686gh2expression of gRNA targeting GALNT2
106687gh3expression of gRNA targeting GALNT3
106688gh4expression of gRNA targeting GALNT4
106689gh5expression of gRNA targeting GALNT5
106690gh6expression of gRNA targeting GALNT6
106692gh8expression of gRNA targeting GALNT8
106693gh9expression of gRNA targeting GALNT9
106694gh13expression of gRNA targeting GALNT13
106695gh22expression of gRNA targeting COSMC
106696gh23expression of gRNA targeting POMT1, Cas9, and GFP
106697gh24expression of gRNA targeting POMT2
106698gh14expression of gRNA targeting GALNT14
106699gh15expression of gRNA targeting GALNT15
106700gh16expression of gRNA targeting GALNT16
106701gh17expression of gRNA targeting GALNT17
106702gh18expression of gRNA targeting GALNT18
106703gh19expression of gRNA targeting GALNT19
106704gh20expression of gRNA targeting GALNT20
106705gh21expression of gRNA targeting C1GALT1
106706gh25expression of gRNA targeting POMGNT1
106707gh44expression of gRNA targeting ST6GALNAC3
106708gh45expression of gRNA targeting ST6GALNAC4
106709gh46expression of gRNA targeting ST6GALNAC5
106710gh33expression of gRNA targeting B4GALNT3
106711gh34expression of gRNA targeting B4GALNT4
106712gh38expression of gRNA targeting STT3A
106713gh39expression of gRNA targeting STT3B
106714gh40expression of gRNA targeting ST6GAL1
106715gh41expression of gRNA targeting ST6GAL2
106716gh42expression of gRNA targeting ST6GALNAC1
106717gh43expression of gRNA targeting ST6GALNAC2
106718gh47expression of gRNA targeting ST6GALNAC6
106720gh60expression of gRNA targeting B3GAT3
106722gh49expression of gRNA targeting ST3GAL1
106723gh50expression of gRNA targeting ST3GAL2
106724gh51expression of gRNA targeting ST3GAL3
106725gh52expression of gRNA targeting ST3GAL4
106726gh53expression of gRNA targeting ST3GAL5
106727gh54expression of gRNA targeting ST3GAL6
106728gh55expression of gRNA targeting B4GALT7
106730gh62expression of gRNA targeting B4GALT2
106731gh71expression of gRNA targeting B3GNT5
106732gh72expression of gRNA targeting B3GNT6
106733gh73expression of gRNA targeting B3GNT7
106734gh63expression of gRNA targeting B4GALT3
106735gh64expression of gRNA targeting B4GALT4
106736gh65expression of gRNA targeting B4GALT5
106737gh66expression of gRNA targeting B4GALT6
106738gh67expression of gRNA targeting B3GNT1 (B4GAT1)
106739gh68expression of gRNA targeting B3GNT2
106740gh69expression of gRNA targeting B3GNT3
106741gh70expression of gRNA targeting B3GNT4
106742gh74expression of gRNA targeting B3GNT8
106743gh83expression of gRNA targeting MGAT5
106744gh84expression of gRNA targeting MGAT5B
106745gh85expression of gRNA targeting FUT1
106746gh75expression of gRNA targeting B3GNT9
106747gh76expression of gRNA targeting MGAT1
106748gh77expression of gRNA targeting MGAT2
106749gh78expression of gRNA targeting MGAT3
106750gh79expression of gRNA targeting MGAT4A
106751gh80expression of gRNA targeting MGAT4B
106752gh81expression of gRNA targeting MGAT4C
106753gh82expression of gRNA targeting MGAT4D
106754gh86expression of gRNA targeting FUT2
106755gh95expression of gRNA targeting FUT11
106756gh96expression of gRNA targeting POFUT1
106757gh97expression of gRNA targeting POFUT2
106758gh87expression of gRNA targeting FUT3
106759gh88expression of gRNA targeting FUT4
106760gh89expression of gRNA targeting FUT5
106761gh90expression of gRNA targeting FUT6
106762gh91expression of gRNA targeting FUT7
106763gh92expression of gRNA targeting FUT8
106764gh93expression of gRNA targeting FUT9
106765gh94expression of gRNA targeting FUT10
106766gh98expression of gRNA targeting LFNG
106767gh108expression of gRNA targeting B3GALT5
106768gh109expression of gRNA targeting B3GALT6
106769gh110expression of gRNA targeting B3GALTL (B3GLCT)
106770gh99expression of gRNA targeting MFNG
106771gh100expression of gRNA targeting RFNG
106772gh101expression of gRNA targeting POGLUT1
106773gh103expression of gRNA targeting GXYLT2
106774gh104expression of gRNA targeting XXYLT1
106775gh105expression of gRNA targeting B3GALT1
106776gh106expression of gRNA targeting B3GALT2
106777gh107expression of gRNA targeting B3GALT4
106778gh111expression of gRNA targeting B3GALNT1
106779gh120expression of gRNA targeting GCNT7
106780gh121expression of gRNA targeting ST8SIA1
106781gh122expression of gRNA targeting ST8SIA2
106782gh112expression of gRNA targeting B3GALNT2
106783gh113expression of gRNA targeting B4GALNT1
106784gh114expression of gRNA targeting B4GALNT2
106785gh115expression of gRNA targeting GCNT1
106786gh116expression of gRNA targeting GCNT2
106787gh117expression of gRNA targeting GCNT3
106788gh118expression of gRNA targeting GCNT4
106789gh119expression of gRNA targeting GCNT6
106790gh123expression of gRNA targeting ST8SIA3
106791gh132expression of gRNA targeting CHPF2
106792gh133expression of gRNA targeting CHSY1
106793gh134expression of gRNA targeting CHSY3
106794gh124expression of gRNA targeting ST8SIA4
106795gh125expression of gRNA targeting ST8SIA5
106796gh126expression of gRNA targeting ST8SIA6
106797gh127expression of gRNA targeting XYLT1
106798gh128expression of gRNA targeting XYLT2
106799gh129expression of gRNA targeting CSGALNACT1
106800gh130expression of gRNA targeting CSGALNACT2
106801gh131expression of gRNA targeting CHPF
106802gh135expression of gRNA targeting EXT1
106803gh144expression of gRNA targeting LARGE
106804gh145expression of gRNA targeting FKRP
106805gh146expression of gRNA targeting FKTN
106806gh136expression of gRNA targeting EXT2
106807gh137expression of gRNA targeting EXTL1
106808gh138expression of gRNA targeting EXTL2
106809gh139expression of gRNA targeting EXTL3
106810gh140expression of gRNA targeting GBGT1
106811gh141expression of gRNA targeting OGT
106812gh142expression of gRNA targeting EOGT
106813gh143expression of gRNA targeting POMGNT2
106814gh147expression of gRNA targeting KDELC1
106815gh162expression of gRNA targeting TMTC3
106816gh163expression of gRNA targeting TMTC4
106817gh166expression of gRNA targeting A3GALT2
106818gh148expression of gRNA targeting KDELC2
106819gh149expression of gRNA targeting GLT1D1
106820gh150expression of gRNA targeting GTDC1
106821gh151expression of gRNA targeting GLT8D1
106822gh152expression of gRNA targeting GLT8D2
106823gh156expression of gRNA targeting GYLT1B (LARGE2)
106824gh160expression of gRNA targeting TMTC1
106825gh161expression of gRNA targeting TMTC2
106826gh167expression of gRNA targeting A4GALT
106827gh177expression of gRNA targeting HAS3
106828gh178expression of gRNA targeting GYS1
106830gh168expression of gRNA targeting A4GNT
106831gh169expression of gRNA targeting ABO
106832gh171expression of gRNA targeting UGCG
106833gh172expression of gRNA targeting UGGT1
106834gh173expression of gRNA targeting UGGT2
106835gh174expression of gRNA targeting UGT8
106836gh175expression of gRNA targeting HAS1
106837gh176expression of gRNA targeting HAS2
106838gh180expression of gRNA targeting GYG1
106839gh236expression of gRNA targeting DPY19L3
106840gh237expression of gRNA targeting DPY19L4
106841gh238expression of gRNA targeting DPM1
106842gh181expression of gRNA targeting GYG2
106843gh182expression of gRNA targeting GLT6D1
106844gh183expression of gRNA targeting CERCAM
106845gh184expression of gRNA targeting COLGALT1
106846gh185expression of gRNA targeting COLGALT2
106847gh186expression of gRNA targeting B3GNTL1
106848gh234expression of gRNA targeting DPY19L1
106849gh235expression of gRNA targeting DPY19L2
106850gh242expression of gRNA targeting PIGA
106851gh260expression of gRNA targeting ALG6
106852gh261expression of gRNA targeting ALG8
106853gh262expression of gRNA targeting ALG9
106854gh243expression of gRNA targeting PIGB
106855gh246expression of gRNA targeting PIGM
106856gh249expression of gRNA targeting PIGV
106857gh250expression of gRNA targeting PIGZ
106859gh257expression of gRNA targeting ALG2
106860gh258expression of gRNA targeting ALG3
106861gh259expression of gRNA targeting ALG5
106862gh263expression of gRNA targeting ALG10
106863gh264expression of gRNA targeting ALG10B
106864gh265expression of gRNA targeting ALG11
106865gh266expression of gRNA targeting ALG12
106866gh267expression of gRNA targeting ALG13
106867gh268expression of gRNA targeting ALG14

Antibodies from Article