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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  
106682gh1expresssion of gRNA targeting GALNT1
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106683gh10expresssion of gRNA targeting GALNT10
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106684gh11expresssion of gRNA targeting GALNT11
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106685gh12expresssion of gRNA targeting GALNT12
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106686gh2expresssion of gRNA targeting GALNT2
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106687gh3expresssion of gRNA targeting GALNT3
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106688gh4expresssion of gRNA targeting GALNT4
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106689gh5expresssion of gRNA targeting GALNT5
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106690gh6expresssion of gRNA targeting GALNT6
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106692gh8expresssion of gRNA targeting GALNT8
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106693gh9expresssion of gRNA targeting GALNT9
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106694gh13expresssion of gRNA targeting GALNT13
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106695gh22expresssion of gRNA targeting COSMC
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106696gh23expresssion of gRNA targeting POMT1, Cas9, and GFP
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106697gh24expresssion of gRNA targeting POMT2
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106698gh14expresssion of gRNA targeting GALNT14
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106699gh15expresssion of gRNA targeting GALNT15
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106700gh16expression of gRNA targeting GALNT16
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106701gh17expresssion of gRNA targeting GALNT17
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106702gh18expresssion of gRNA targeting GALNT18
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106703gh19expresssion of gRNA targeting GALNT19
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106704gh20expresssion of gRNA targeting GALNT20
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106705gh21expresssion of gRNA targeting C1GALT1
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106706gh25expresssion of gRNA targeting POMGNT1
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106707gh44expresssion of gRNA targeting ST6GALNAC3
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106708gh45expresssion of gRNA targeting ST6GALNAC4
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106709gh46expresssion of gRNA targeting ST6GALNAC5
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106710gh33expresssion of gRNA targeting B4GALNT3
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106711gh34expresssion of gRNA targeting B4GALNT4
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106712gh38expresssion of gRNA targeting STT3A
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106713gh39expresssion of gRNA targeting STT3B
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106714gh40expresssion of gRNA targeting ST6GAL1
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106715gh41expresssion of gRNA targeting ST6GAL2
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106716gh42expresssion of gRNA targeting ST6GALNAC1
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106717gh43expresssion of gRNA targeting ST6GALNAC2
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106718gh47expresssion of gRNA targeting ST6GALNAC6
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106720gh60expresssion of gRNA targeting B3GAT3
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106722gh49expresssion of gRNA targeting ST3GAL1
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106723gh50expresssion of gRNA targeting ST3GAL2
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106724gh51expresssion of gRNA targeting ST3GAL3
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106725gh52expresssion of gRNA targeting ST3GAL4
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106726gh53expresssion of gRNA targeting ST3GAL5
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106727gh54expresssion of gRNA targeting ST3GAL6
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106728gh55expresssion of gRNA targeting B4GALT7
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106730gh62expresssion of gRNA targeting B4GALT2
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106731gh71expresssion of gRNA targeting B3GNT5
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106732gh72expresssion of gRNA targeting B3GNT6
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106733gh73expresssion of gRNA targeting B3GNT7
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106734gh63expresssion of gRNA targeting B4GALT3
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106735gh64expression of gRNA targeting B4GALT4
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106736gh65expresssion of gRNA targeting B4GALT5
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106737gh66expresssion of gRNA targeting B4GALT6
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106738gh67expresssion of gRNA targeting B3GNT1 (B4GAT1)
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106739gh68expresssion of gRNA targeting B3GNT2
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106740gh69expresssion of gRNA targeting B3GNT3
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106741gh70expresssion of gRNA targeting B3GNT4
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106742gh74expresssion of gRNA targeting B3GNT8
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106743gh83expresssion of gRNA targeting MGAT5
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106744gh84expresssion of gRNA targeting MGAT5B
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106745gh85expresssion of gRNA targeting FUT1
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106746gh75expresssion of gRNA targeting B3GNT9
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106747gh76expresssion of gRNA targeting MGAT1
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106748gh77expresssion of gRNA targeting MGAT2
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106749gh78expresssion of gRNA targeting MGAT3
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106750gh79expresssion of gRNA targeting MGAT4A
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106751gh80expresssion of gRNA targeting MGAT4B
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106752gh81expresssion of gRNA targeting MGAT4C
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106753gh82expresssion of gRNA targeting MGAT4D
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106754gh86expresssion of gRNA targeting FUT2
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106755gh95expresssion of gRNA targeting FUT11
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106756gh96expresssion of gRNA targeting POFUT1
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106757gh97expresssion of gRNA targeting POFUT2
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106758gh87expresssion of gRNA targeting FUT3
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106759gh88expresssion of gRNA targeting FUT4
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106760gh89expresssion of gRNA targeting FUT5
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106761gh90expresssion of gRNA targeting FUT6
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106762gh91expresssion of gRNA targeting FUT7
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106763gh92expresssion of gRNA targeting FUT8
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106764gh93expresssion of gRNA targeting FUT9
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106765gh94expresssion of gRNA targeting FUT10
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106766gh98expresssion of gRNA targeting LFNG
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106767gh108expresssion of gRNA targeting B3GALT5
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106768gh109expresssion of gRNA targeting B3GALT6
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106769gh110expresssion of gRNA targeting B3GALTL (B3GLCT)
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106770gh99expresssion of gRNA targeting MFNG
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106771gh100expresssion of gRNA targeting RFNG
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106772gh101expresssion of gRNA targeting POGLUT1
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106773gh103expresssion of gRNA targeting GXYLT2
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106774gh104expresssion of gRNA targeting XXYLT1
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106775gh105expresssion of gRNA targeting B3GALT1
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106776gh106expresssion of gRNA targeting B3GALT2
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106777gh107expresssion of gRNA targeting B3GALT4
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106778gh111expresssion of gRNA targeting B3GALNT1
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106779gh120expresssion of gRNA targeting GCNT7
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106780gh121expresssion of gRNA targeting ST8SIA1
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106781gh122expresssion of gRNA targeting ST8SIA2
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106782gh112expresssion of gRNA targeting B3GALNT2
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106783gh113expresssion of gRNA targeting B4GALNT1
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106784gh114expresssion of gRNA targeting B4GALNT2
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106785gh115expresssion of gRNA targeting GCNT1
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106786gh116expression of gRNA targeting GCNT2
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106787gh117expresssion of gRNA targeting GCNT3
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106788gh118expresssion of gRNA targeting GCNT4
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106789gh119expresssion of gRNA targeting GCNT6
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106790gh123expresssion of gRNA targeting ST8SIA3
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106791gh132expresssion of gRNA targeting CHPF2
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106792gh133expresssion of gRNA targeting CHSY1
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106793gh134expresssion of gRNA targeting CHSY3
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106794gh124expresssion of gRNA targeting ST8SIA4
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106795gh125expresssion of gRNA targeting ST8SIA5
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106796gh126expresssion of gRNA targeting ST8SIA6
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106797gh127expresssion of gRNA targeting XYLT1
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106798gh128expresssion of gRNA targeting XYLT2
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106799gh129expresssion of gRNA targeting CSGALNACT1
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106800gh130expresssion of gRNA targeting CSGALNACT2
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106801gh131expresssion of gRNA targeting CHPF
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106802gh135expresssion of gRNA targeting EXT1
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106803gh144expresssion of gRNA targeting LARGE
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106804gh145expresssion of gRNA targeting FKRP
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106805gh146expresssion of gRNA targeting FKTN
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106806gh136expresssion of gRNA targeting EXT2
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106807gh137expresssion of gRNA targeting EXTL1
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106808gh138expresssion of gRNA targeting EXTL2
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106809gh139expresssion of gRNA targeting EXTL3
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106810gh140expresssion of gRNA targeting GBGT1
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106811gh141expresssion of gRNA targeting OGT
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106812gh142expresssion of gRNA targeting EOGT
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106813gh143expresssion of gRNA targeting POMGNT2
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106814gh147expresssion of gRNA targeting KDELC1
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106815gh162expresssion of gRNA targeting TMTC3
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106816gh163expresssion of gRNA targeting TMTC4
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106817gh166expresssion of gRNA targeting A3GALT2
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106818gh148expresssion of gRNA targeting KDELC2
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106819gh149expresssion of gRNA targeting GLT1D1
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106820gh150expresssion of gRNA targeting GTDC1
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106821gh151expresssion of gRNA targeting GLT8D1
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106822gh152expresssion of gRNA targeting GLT8D2
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106823gh156expresssion of gRNA targeting GYLT1B (LARGE2)
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106824gh160expresssion of gRNA targeting TMTC1
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106825gh161expresssion of gRNA targeting TMTC2
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106826gh167expresssion of gRNA targeting A4GALT
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106827gh177expresssion of gRNA targeting HAS3
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106828gh178expresssion of gRNA targeting GYS1
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106830gh168expresssion of gRNA targeting A4GNT
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106831gh169expresssion of gRNA targeting ABO
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106832gh171expresssion of gRNA targeting UGCG
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106833gh172expresssion of gRNA targeting UGGT1
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106834gh173expresssion of gRNA targeting UGGT2
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106835gh174expresssion of gRNA targeting UGT8
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106836gh175expresssion of gRNA targeting HAS1
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106837gh176expresssion of gRNA targeting HAS2
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106838gh180expresssion of gRNA targeting GYG1
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106839gh236expresssion of gRNA targeting DPY19L3
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106840gh237expresssion of gRNA targeting DPY19L4
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106841gh238expresssion of gRNA targeting DPM1
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106842gh181expresssion of gRNA targeting GYG2
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106843gh182expresssion of gRNA targeting GLT6D1
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106844gh183expresssion of gRNA targeting CERCAM
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106845gh184expresssion of gRNA targeting COLGALT1
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106846gh185expresssion of gRNA targeting COLGALT2
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106847gh186expresssion of gRNA targeting B3GNTL1
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106848gh234expresssion of gRNA targeting DPY19L1
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106849gh235expresssion of gRNA targeting DPY19L2
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106850gh242expresssion of gRNA targeting PIGA
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106851gh260expresssion of gRNA targeting ALG6
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106852gh261expresssion of gRNA targeting ALG8
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106853gh262expresssion of gRNA targeting ALG9
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106854gh243expresssion of gRNA targeting PIGB
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106855gh246expresssion of gRNA targeting PIGM
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106856gh249expresssion of gRNA targeting PIGV
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106857gh250expresssion of gRNA targeting PIGZ
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106859gh257expresssion of gRNA targeting ALG2
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106860gh258expresssion of gRNA targeting ALG3
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106861gh259expresssion of gRNA targeting ALG5
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106862gh263expresssion of gRNA targeting ALG10
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106863gh264expresssion of gRNA targeting ALG10B
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106864gh265expresssion of gRNA targeting ALG11
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106865gh266expresssion of gRNA targeting ALG12
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106866gh267expresssion of gRNA targeting ALG13
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106867gh268expresssion of gRNA targeting ALG14
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Recombinant Antibodies from Article