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