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.
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PubMed
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Article
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 KBAdditional 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 |
---|---|---|
106682 | gh1 | expression of gRNA targeting GALNT1 |
106683 | gh10 | expression of gRNA targeting GALNT10 |
106684 | gh11 | expression of gRNA targeting GALNT11 |
106685 | gh12 | expression of gRNA targeting GALNT12 |
106686 | gh2 | expression of gRNA targeting GALNT2 |
106687 | gh3 | expression of gRNA targeting GALNT3 |
106688 | gh4 | expression of gRNA targeting GALNT4 |
106689 | gh5 | expression of gRNA targeting GALNT5 |
106690 | gh6 | expression of gRNA targeting GALNT6 |
106692 | gh8 | expression of gRNA targeting GALNT8 |
106693 | gh9 | expression of gRNA targeting GALNT9 |
106694 | gh13 | expression of gRNA targeting GALNT13 |
106695 | gh22 | expression of gRNA targeting COSMC |
106696 | gh23 | expression of gRNA targeting POMT1, Cas9, and GFP |
106697 | gh24 | expression of gRNA targeting POMT2 |
106698 | gh14 | expression of gRNA targeting GALNT14 |
106699 | gh15 | expression of gRNA targeting GALNT15 |
106700 | gh16 | expression of gRNA targeting GALNT16 |
106701 | gh17 | expression of gRNA targeting GALNT17 |
106702 | gh18 | expression of gRNA targeting GALNT18 |
106703 | gh19 | expression of gRNA targeting GALNT19 |
106704 | gh20 | expression of gRNA targeting GALNT20 |
106705 | gh21 | expression of gRNA targeting C1GALT1 |
106706 | gh25 | expression of gRNA targeting POMGNT1 |
106707 | gh44 | expression of gRNA targeting ST6GALNAC3 |
106708 | gh45 | expression of gRNA targeting ST6GALNAC4 |
106709 | gh46 | expression of gRNA targeting ST6GALNAC5 |
106710 | gh33 | expression of gRNA targeting B4GALNT3 |
106711 | gh34 | expression of gRNA targeting B4GALNT4 |
106712 | gh38 | expression of gRNA targeting STT3A |
106713 | gh39 | expression of gRNA targeting STT3B |
106714 | gh40 | expression of gRNA targeting ST6GAL1 |
106715 | gh41 | expression of gRNA targeting ST6GAL2 |
106716 | gh42 | expression of gRNA targeting ST6GALNAC1 |
106717 | gh43 | expression of gRNA targeting ST6GALNAC2 |
106718 | gh47 | expression of gRNA targeting ST6GALNAC6 |
106720 | gh60 | expression of gRNA targeting B3GAT3 |
106722 | gh49 | expression of gRNA targeting ST3GAL1 |
106723 | gh50 | expression of gRNA targeting ST3GAL2 |
106724 | gh51 | expression of gRNA targeting ST3GAL3 |
106725 | gh52 | expression of gRNA targeting ST3GAL4 |
106726 | gh53 | expression of gRNA targeting ST3GAL5 |
106727 | gh54 | expression of gRNA targeting ST3GAL6 |
106728 | gh55 | expression of gRNA targeting B4GALT7 |
106730 | gh62 | expression of gRNA targeting B4GALT2 |
106731 | gh71 | expression of gRNA targeting B3GNT5 |
106732 | gh72 | expression of gRNA targeting B3GNT6 |
106733 | gh73 | expression of gRNA targeting B3GNT7 |
106734 | gh63 | expression of gRNA targeting B4GALT3 |
106735 | gh64 | expression of gRNA targeting B4GALT4 |
106736 | gh65 | expression of gRNA targeting B4GALT5 |
106737 | gh66 | expression of gRNA targeting B4GALT6 |
106738 | gh67 | expression of gRNA targeting B3GNT1 (B4GAT1) |
106739 | gh68 | expression of gRNA targeting B3GNT2 |
106740 | gh69 | expression of gRNA targeting B3GNT3 |
106741 | gh70 | expression of gRNA targeting B3GNT4 |
106742 | gh74 | expression of gRNA targeting B3GNT8 |
106743 | gh83 | expression of gRNA targeting MGAT5 |
106744 | gh84 | expression of gRNA targeting MGAT5B |
106745 | gh85 | expression of gRNA targeting FUT1 |
106746 | gh75 | expression of gRNA targeting B3GNT9 |
106747 | gh76 | expression of gRNA targeting MGAT1 |
106748 | gh77 | expression of gRNA targeting MGAT2 |
106749 | gh78 | expression of gRNA targeting MGAT3 |
106750 | gh79 | expression of gRNA targeting MGAT4A |
106751 | gh80 | expression of gRNA targeting MGAT4B |
106752 | gh81 | expression of gRNA targeting MGAT4C |
106753 | gh82 | expression of gRNA targeting MGAT4D |
106754 | gh86 | expression of gRNA targeting FUT2 |
106755 | gh95 | expression of gRNA targeting FUT11 |
106756 | gh96 | expression of gRNA targeting POFUT1 |
106757 | gh97 | expression of gRNA targeting POFUT2 |
106758 | gh87 | expression of gRNA targeting FUT3 |
106759 | gh88 | expression of gRNA targeting FUT4 |
106760 | gh89 | expression of gRNA targeting FUT5 |
106761 | gh90 | expression of gRNA targeting FUT6 |
106762 | gh91 | expression of gRNA targeting FUT7 |
106763 | gh92 | expression of gRNA targeting FUT8 |
106764 | gh93 | expression of gRNA targeting FUT9 |
106765 | gh94 | expression of gRNA targeting FUT10 |
106766 | gh98 | expression of gRNA targeting LFNG |
106767 | gh108 | expression of gRNA targeting B3GALT5 |
106768 | gh109 | expression of gRNA targeting B3GALT6 |
106769 | gh110 | expression of gRNA targeting B3GALTL (B3GLCT) |
106770 | gh99 | expression of gRNA targeting MFNG |
106771 | gh100 | expression of gRNA targeting RFNG |
106772 | gh101 | expression of gRNA targeting POGLUT1 |
106773 | gh103 | expression of gRNA targeting GXYLT2 |
106774 | gh104 | expression of gRNA targeting XXYLT1 |
106775 | gh105 | expression of gRNA targeting B3GALT1 |
106776 | gh106 | expression of gRNA targeting B3GALT2 |
106777 | gh107 | expression of gRNA targeting B3GALT4 |
106778 | gh111 | expression of gRNA targeting B3GALNT1 |
106779 | gh120 | expression of gRNA targeting GCNT7 |
106780 | gh121 | expression of gRNA targeting ST8SIA1 |
106781 | gh122 | expression of gRNA targeting ST8SIA2 |
106782 | gh112 | expression of gRNA targeting B3GALNT2 |
106783 | gh113 | expression of gRNA targeting B4GALNT1 |
106784 | gh114 | expression of gRNA targeting B4GALNT2 |
106785 | gh115 | expression of gRNA targeting GCNT1 |
106786 | gh116 | expression of gRNA targeting GCNT2 |
106787 | gh117 | expression of gRNA targeting GCNT3 |
106788 | gh118 | expression of gRNA targeting GCNT4 |
106789 | gh119 | expression of gRNA targeting GCNT6 |
106790 | gh123 | expression of gRNA targeting ST8SIA3 |
106791 | gh132 | expression of gRNA targeting CHPF2 |
106792 | gh133 | expression of gRNA targeting CHSY1 |
106793 | gh134 | expression of gRNA targeting CHSY3 |
106794 | gh124 | expression of gRNA targeting ST8SIA4 |
106795 | gh125 | expression of gRNA targeting ST8SIA5 |
106796 | gh126 | expression of gRNA targeting ST8SIA6 |
106797 | gh127 | expression of gRNA targeting XYLT1 |
106798 | gh128 | expression of gRNA targeting XYLT2 |
106799 | gh129 | expression of gRNA targeting CSGALNACT1 |
106800 | gh130 | expression of gRNA targeting CSGALNACT2 |
106801 | gh131 | expression of gRNA targeting CHPF |
106802 | gh135 | expression of gRNA targeting EXT1 |
106803 | gh144 | expression of gRNA targeting LARGE |
106804 | gh145 | expression of gRNA targeting FKRP |
106805 | gh146 | expression of gRNA targeting FKTN |
106806 | gh136 | expression of gRNA targeting EXT2 |
106807 | gh137 | expression of gRNA targeting EXTL1 |
106808 | gh138 | expression of gRNA targeting EXTL2 |
106809 | gh139 | expression of gRNA targeting EXTL3 |
106810 | gh140 | expression of gRNA targeting GBGT1 |
106811 | gh141 | expression of gRNA targeting OGT |
106812 | gh142 | expression of gRNA targeting EOGT |
106813 | gh143 | expression of gRNA targeting POMGNT2 |
106814 | gh147 | expression of gRNA targeting KDELC1 |
106815 | gh162 | expression of gRNA targeting TMTC3 |
106816 | gh163 | expression of gRNA targeting TMTC4 |
106817 | gh166 | expression of gRNA targeting A3GALT2 |
106818 | gh148 | expression of gRNA targeting KDELC2 |
106819 | gh149 | expression of gRNA targeting GLT1D1 |
106820 | gh150 | expression of gRNA targeting GTDC1 |
106821 | gh151 | expression of gRNA targeting GLT8D1 |
106822 | gh152 | expression of gRNA targeting GLT8D2 |
106823 | gh156 | expression of gRNA targeting GYLT1B (LARGE2) |
106824 | gh160 | expression of gRNA targeting TMTC1 |
106825 | gh161 | expression of gRNA targeting TMTC2 |
106826 | gh167 | expression of gRNA targeting A4GALT |
106827 | gh177 | expression of gRNA targeting HAS3 |
106828 | gh178 | expression of gRNA targeting GYS1 |
106830 | gh168 | expression of gRNA targeting A4GNT |
106831 | gh169 | expression of gRNA targeting ABO |
106832 | gh171 | expression of gRNA targeting UGCG |
106833 | gh172 | expression of gRNA targeting UGGT1 |
106834 | gh173 | expression of gRNA targeting UGGT2 |
106835 | gh174 | expression of gRNA targeting UGT8 |
106836 | gh175 | expression of gRNA targeting HAS1 |
106837 | gh176 | expression of gRNA targeting HAS2 |
106838 | gh180 | expression of gRNA targeting GYG1 |
106839 | gh236 | expression of gRNA targeting DPY19L3 |
106840 | gh237 | expression of gRNA targeting DPY19L4 |
106841 | gh238 | expression of gRNA targeting DPM1 |
106842 | gh181 | expression of gRNA targeting GYG2 |
106843 | gh182 | expression of gRNA targeting GLT6D1 |
106844 | gh183 | expression of gRNA targeting CERCAM |
106845 | gh184 | expression of gRNA targeting COLGALT1 |
106846 | gh185 | expression of gRNA targeting COLGALT2 |
106847 | gh186 | expression of gRNA targeting B3GNTL1 |
106848 | gh234 | expression of gRNA targeting DPY19L1 |
106849 | gh235 | expression of gRNA targeting DPY19L2 |
106850 | gh242 | expression of gRNA targeting PIGA |
106851 | gh260 | expression of gRNA targeting ALG6 |
106852 | gh261 | expression of gRNA targeting ALG8 |
106853 | gh262 | expression of gRNA targeting ALG9 |
106854 | gh243 | expression of gRNA targeting PIGB |
106855 | gh246 | expression of gRNA targeting PIGM |
106856 | gh249 | expression of gRNA targeting PIGV |
106857 | gh250 | expression of gRNA targeting PIGZ |
106859 | gh257 | expression of gRNA targeting ALG2 |
106860 | gh258 | expression of gRNA targeting ALG3 |
106861 | gh259 | expression of gRNA targeting ALG5 |
106862 | gh263 | expression of gRNA targeting ALG10 |
106863 | gh264 | expression of gRNA targeting ALG10B |
106864 | gh265 | expression of gRNA targeting ALG11 |
106865 | gh266 | expression of gRNA targeting ALG12 |
106866 | gh267 | expression of gRNA targeting ALG13 |
106867 | gh268 | expression of gRNA targeting ALG14 |