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THAT HAD TORTE I USUNA 20180016601A1ALTRA MANIAN LITERATURA UNA ( 19) United States (12 ) Patent Application Publication ( 10) Pub . No. : US 2018 / 0016601 A1 Qi et al. ( 43 ) Pub . Date: Jan . 18 , 2018 ( 54 ) METHODS FOR MODULATING GENOME C12N 15 / 10 (2006 .01 ) EDITING A6IK 38 / 46 ( 2006 .01 ) A61K 31 /365 (2006 .01 ) ( 71) Applicants: The Board of Trustees of the Leland A61K 31/ 63 ( 2006 .01 ) Stanford Junior University , Palo Alto , A61K 31 /513 ( 2006 . 01 ) CA (US ) ; The J . David Gladstone A61K 31 /505 ( 2006 .01 ) Institutes , a Testamentary Trust C12Q 1 / 68 ( 2006 .01 ) established under the Will of J . David C12N 9 / 22 ( 2006 . 01 ) Glads, San Francisco , CA ( US) ; The (52 ) U . S . CI. Regents of the University of CPC . .. - . C12N 15/ 907 ( 2013 .01 ) ; C12Q 1 /68 California , Oakland , CA (US ) ( 2013 . 01 ) ; C12Q 1 /44 ( 2013 .01 ) ; C12N 15 / 1024 (2013 .01 ) ; C12N 9 /22 ( 2013. 01 ) ; ( 72 ) Inventors : Lei S . Qi, Palo Alto , CA (US ) ; Sheng A61K 31/ 365 ( 2013 . 01) ; A61K 31 /63 Ding , Orinda , CA ( US ) ; Chen Yu , San ( 2013 .01 ) ; A61K 31/ 513 ( 2013 . 01 ) ; A61K Francisco , CA (US ) 31/ 505 ( 2013. 01 ) ; A61K 38 /465 (2013 . 01 ) (57 ) ABSTRACT ( 21 ) Appl. No. : 15 / 649, 304 Provided herein are methods and kits for modulating (22 ) Filed : Jul. 13 , 2017 genome editing of target DNA . The invention includes using small molecules that enhance or repress homology - directed Related U . S . Application Data repair (HDR ) and / or nonhomologous end joining (NHEJ ) (63 ) Continuation of application No . PCT /US2016 / repair of double - strand breaks in a target DNA sequence . 013375 , filed on Jan . 14 , 2016 . Also provided herein are methods for preventing or treating (60 ) Provisional application No . 62 / 104, 035 , filed on Jan . a genetic disease in a subject by enhancing precise genome 15 , 2015 . editing to correct a mutation in a target gene associated with the genetic disease . Further provided herein are systems and Publication Classification methods for screening small molecule libraries to identify (51 ) Int . Cl. novel modulators of genome editing . The present invention C12N 15 / 90 (2006 .01 ) can be used with any cell type and at any gene locus that is C12Q 1 /44 ( 2006 .01 ) amenable to nuclease -mediated genome editing technology . Patent Application Publication Jan . 18 , 2018 Sheet 1 of 13 US 2018 /0016601 A1 SgNanog 0% template 0.05%. 000000000000000000000000000000000000000 FIG.10 *Caso SgNanog 16.79% iFi SgNanog template 0.26% ensdeconsumoosobnossono * wwwwwww????? FIG.1B ZZZZ X2404040404oco440000000000ococo400000000000ococo40000004cococo Vzd Nanog3'HA neprimerset1 Primersat2 togta5: ascati G winniin SORNAtargetsite www SCHNLO2A- Loftarm(1.8kb)Mightam24 STOFFPLS2A- ML 34horas HOR WOWILS 230 2)DI Mathimit PAM Nanog5'HA SfGFP ECIOCACCAGOR3GAGGTGONDOL insomethinnainen FIG.1A Nanog I. Nanog CCACAAGCCTTGGAATTATTCCTGAACTACTCTGTGACTCCACCAGGTGAAATAGCTACTAACTTCAGCCTGCTGA GAGTTTGTAACTGCTGCTGGGATTACACATGGCATGGATGAGCTCTACAAATGAGACTTACGCAACATCTGGGCITZZZZZZZZZZZZZZZZZZZZZZZZ Patent Application Publication Jan . 18 , 2018 Sheet 2 of 13 US 2018 /0016601 A1 5.9ko5.0kb 3.4kb Jadua: 4,000 96 2 Mean 884 AZT AETA ABrefeldin '. TUE 185241|1 L755507\ Statisticsofsequencedalleles 19296 2,000 Jayquapipunodwoo DMSO 614 0 98 Brefeldinu .6756507 primerset1 primerset2 Chr19 Correct{nsert Alleles Sequenced 1F.FIG 08 OC OLO Bonu + d29 % 6.78% TFT 6,05% cellsCulture w Valdatein12wellplates Seedcells Scanandanalyze + Dectroporation 33.3% BrefeldinA 27.2% L765507 000000000 w 177%- Escels 0% 384-wellplate DMSO Addcompounds Veveria Www SIGFP 00000000000 Counts. ,FIG.1D H FIG.1E Patent Application Publication Jan . 18 , 2018 Sheet 3 of 13 US 2018 / 0016601 A1 www OH ww. Concentration(M) 5.1001 W MO wWww wWw wWw w 0.25 * * * * Mais * * Concentration(UM) 5.1001 * ome * H * 1.00-* ullall0.75 0.50 0.25 Concentration(UM) ho BrefeldinA 001010.105' 2.5 Hey hey 1,5 02 bogen-1. Concentration(M) t755507Alt 0.0115 FIG.1G OSWO 01 paziewou Sie d0 % Patent Application Publication Jan . 18 , 2018 Sheet 4 of 13 US 2018 / 0016601 A1 FIG . 2A PAM SORNA target site 5 * GAAGccGGGccTTCCATTGTCCACCGCAAATGCT 3 * *3 ' CTTCGGCCCGGAAGGTAACAGGTGGCGTTTACGA 5 ! - 8 - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - - . ????2 12A - Venus Left arm (750 bp ) Right arm (675 bp ) HDR t2A - Venus - B 9 ACTA2 Patent Application Publication Jan . 18 , 2018 Sheet 5 of 13 US 2018 / 0016601 A1 ?????????????????? t2A ACTA23'HA E????????????????????????????????. ?????????????????????????????????GCCGCCGGGATCACTUTCGCCATGGACGAGCTOTACAAGTAATAGGGTACCACTITCCTOc?? FIG.2B ACTA25'HA Venus GGGCTCCATTGTCCACCGCAAATGCTTCGGTACCAGCGGCAGCGGAGAGGGCAGAGGAAGTCTTCTAACATGCGGTGACGTGGAGGAGAATCCC . Patent Application Publication Jan . 18 , 2018 Sheet 6 of 13 US 2018 /0016601 A1 PAMSORNAtargetsite wildtype5:GAAGGCCGZETEICOLELAGCSEREGGCC3*SOD13'CTTCCGGCAACCORDANODOSLOGG5* FIG,2D SSOON template WW A4VmutationGTC wildtypeSOD1GCC DMSODMSQ coilstem neural 01755507 X drived-hESCs HUVEC * Fibroblast)2097 -CRL ( K562 FIG.2E FIG,2C Hela SNURA * * ** * * * * * * * * * * * * CCTCGGAACCAGGACCTCGGCGTGGCCTAGCGAGTTATGGCGACGAAGGTCGTGTGCGTGCTGAAGGGCGACGGCCCAGTGCAGGGCATCATCAAT Patent Application Publication Jan . 18 , 2018 Sheet 7 of 13 US 2018 /0016601 A1 AZT 4. * * * * * * * * * * * * * * * * * * * * * ** * * * ** * * ** * * * * * * * ** ** * * ** * ** * * * * * * * * * * * * * * * * * ** * * Eddoos Sexy 1755507 2987 %260 Cd1905 44444 wwwwwwwwwwwwww 4 - 6 - 764444444 - X , With 2-300I DMSO W0 . 90 999999999 , , , , , , , , 44444444444444444444444 wwwwwwwwwwwwwwwwww FL2 1908 Zdoos Ed 90s 1904 ????????????????????????????? Nanog-sfGFP EScells Nanog 0.41% FIG.2G chools SOGAL4 0.35%(1/288) 3.13%(6/192) 33.3%(32/96) 31.9%(92/288) 27.6%(53/192) 4 40 frequency(26) 3 Indetallelefrequency(%) 30 44Vmutantallele 2 20 1 10 0 0 = {? ????? FIG.2F L755507 Notemplate 209994 }{q Patent Application Publication Jan . 18 , 2018 Sheet 8 of 13 US 2018/ 0016601 A1 Validateusing12-wellplates ? "4 -4 ?? " - INCellAnalyzer* - ? Achemicalscreeningplatformforgenomeinsertion 3daysofculture 1 AddcompoundsSeedcells(2,000/well)Scanandanalyze "h Cas9+SERNAtemplate Electroporation "?? ) cellsEsmouse3x10 ?????????447hh BrefeldinA 3rFle. "4h h ? - « ?F NanogHAS 101102103104 ??????????? -??? 77-44 { ??????????? 2448hh ????????????? -*}{at 101102103104 ??- FIG,3c ? ? ? ? FIG.3A {} ??? FF Patent Application Publication Jan . 18 , 2018 Sheet 9 of 13 US 2018 /0016601 A1 TFT AZT ho L755507DMSO** template*No*** * Cells E14 to normalized m Viability Cell % FIG.3E TFT AZThtttt ABrefeldin11 L755507DMSO oo om templateNo 6 )' 10 * ( Number Cell FIG.3D Patent Application Publication Jan . 18 , 2018 Sheet 10 of 13 US 2018 /0016601 A1 yurish 5:8Kb 3.4kb ngguna104 cellsES 101102103104 103 84.52% 755507 8424% sfGFP-Nanog OMSO Swapumpunganpe101102 cellsES 104 103 Isotype *107102103104 AZT 85.11 wayanngayongtao101102 FIG.4B Primerset1 Primerset2 Nanog FIG.4D oooooooooooooooooooooooooooooooooooooo Count DAPI Resistant Rightarm 2000000000000000000000000000000000000000000000000 Sox2 1 02AWL6SICFO comam 00000 DAPI 2. 29 Sena Oct4 Vanogallele1 Nanogallele2 Primerset2 AZT FIG.4A 000000000000000000000000000000000 FIG.4C L755507 X900 Patent Application Publication Jan . 18 , 2018 Sheet 11 of 13 US 2018 /0016601 A1 Merge DAP! SGFP Nanog DMSO L755507 FIG.44 AZT Merge DAPI SGFP Nanog AZT cels ES 1 SGGFP 2 - SgGFP 3- SgGFP FIG.4E sfGFP- Nanog FIG.4G Patent Application Publication Jan . 18 , 2018 Sheet 12 of 13 US 2018 /0016601 A1 FIG . 5 OMO L755507 ZT winninneneinaniniwinteiiniiniiiiiiiiiiiiiiiiiiiiiiiiiiiiiiiindet mutaion Total reads 24. 998149 ,211 21. 313: 57 202 49 ,067 /63 , 075 Indel 38 50 . 80 % 47 74 % maininnnnnnnnnn ws un59 ,06 % Patent Application Publication Jan . 18 , 2018 Sheet 13 of 13 US 2018 /0016601 A1 No HR No compound * * * * * * * * * * * dog amentinin n ??????????????????????????????????????????????????? 9994444444444 0 . 19 + 0 .09 wanamtamatondobo 9 . 16 # 1 .85 whiratininwinninanaiskonsantren pratamata Winning L755507 SCR7a L755507 + SCR7a : Meeresbian ???????????????????? : www : OSS 14 .67 + 0 .42 14 .7310 .40 melder 21. 90 + 1 .67 hommes times lacouturistiwatiwww itulacionsionistiwww down phomanswww whatsside GFP FIG . 6 US 2018 / 0016601 A1 Jan . 18 , 2018 METHODS FOR MODULATING GENOME BRIEF SUMMARY OF THE INVENTION EDITING 0005 ] The present invention provides methods and kits for modulating genome editing of target DNA . The inven CROSS -REFERENCES TO RELATED tion includes using small molecules that enhance or repress APPLICATIONS homology - directed repair (HDR ) and / or nonhomologous [ 0001 ] The present application is a Continuation of PCT/ end joining (NHEJ ) repair of double - strand breaks in a target US2016 /013375 filed Jan . 14 , 2016 ; which claims priority to DNA sequence . The present invention also provides meth ods for preventing or treating a disease in a subject by U . S . Provisional Patent Application No . 62 / 104 ,035 filed enhancing precise genome editing to correct a mutation in a Jan . 15 , 2015 ; the disclosures which are hereby incorporated target gene associated with the disease . The present inven by reference in their entirety for all purposes . tion further provides systems and methods for screening small molecule libraries to identify novel modulators of STATEMENT AS TO RIGHTS TO INVENTIONS genome editing . The present invention can be used with any MADE
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    Clenbuterol Elisa Kit Instructions Product #101219 & 101216 Forensic Use Only

    Neogen Corporation 944 Nandino Blvd., Lexington KY 40511 USA 800/477-8201 USA/Canada | 859/254-1221 Fax: 859/255-5532 | E-mail: [email protected] | Web: www.neogen.com/Toxicology CLENBUTEROL ELISA KIT INSTRUCTIONS PRODUCT #101219 & 101216 FORENSIC USE ONLY INTENDED USE: For the determination of trace quantities of Clenbuterol and/or other metabolites in human urine, blood, oral fluid. DESCRIPTION Neogen Corporation’s Clenbuterol ELISA (Enzyme-Linked ImmunoSorbent Assay) test kit is a qualitative one-step kit designed for use as a screening device for the detection of drugs and/or their metabolites. The kit was designed for screening purposes and is intended for forensic use only. It is recommended that all suspect samples be confirmed by a quantitative method such as gas chromatography/mass spectrometry (GC/MS). ASSAY PRINCIPLES Neogen Corporation’s test kit operates on the basis of competition between the drug or its metabolite in the sample and the drug- enzyme conjugate for a limited number of antibody binding sites. First, the sample or control is added to the microplate. Next, the diluted drug-enzyme conjugate is added and the mixture is incubated at room temperature. During this incubation, the drug in the sample or the drug-enzyme conjugate binds to antibody immobilized in the microplate wells. After incubation, the plate is washed 3 times to remove any unbound sample or drug-enzyme conjugate. The presence of bound drug-enzyme conjugate is recognized by the addition of K-Blue® Substrate (TMB). After a 30 minute substrate incubation, the reaction is halted with the addition of Red Stop Solution.
  • The Use of Stems in the Selection of International Nonproprietary Names (INN) for Pharmaceutical Substances

    The Use of Stems in the Selection of International Nonproprietary Names (INN) for Pharmaceutical Substances

    WHO/PSM/QSM/2006.3 The use of stems in the selection of International Nonproprietary Names (INN) for pharmaceutical substances 2006 Programme on International Nonproprietary Names (INN) Quality Assurance and Safety: Medicines Medicines Policy and Standards The use of stems in the selection of International Nonproprietary Names (INN) for pharmaceutical substances FORMER DOCUMENT NUMBER: WHO/PHARM S/NOM 15 © World Health Organization 2006 All rights reserved. Publications of the World Health Organization can be obtained from WHO Press, World Health Organization, 20 Avenue Appia, 1211 Geneva 27, Switzerland (tel.: +41 22 791 3264; fax: +41 22 791 4857; e-mail: [email protected]). Requests for permission to reproduce or translate WHO publications – whether for sale or for noncommercial distribution – should be addressed to WHO Press, at the above address (fax: +41 22 791 4806; e-mail: [email protected]). The designations employed and the presentation of the material in this publication do not imply the expression of any opinion whatsoever on the part of the World Health Organization concerning the legal status of any country, territory, city or area or of its authorities, or concerning the delimitation of its frontiers or boundaries. Dotted lines on maps represent approximate border lines for which there may not yet be full agreement. The mention of specific companies or of certain manufacturers’ products does not imply that they are endorsed or recommended by the World Health Organization in preference to others of a similar nature that are not mentioned. Errors and omissions excepted, the names of proprietary products are distinguished by initial capital letters.
  • CATO佳途科技- Cato Research Chemicals Inc

    CATO佳途科技- Cato Research Chemicals Inc

    CATO 现货标准品产品清单 Cato Research Chemicals Inc. 广州佳途科技股份有限公司 CATO Research Chemicals Inc.专注于标准品研发、生产、服务及销 售，率先在中国建立亚洲技术中心实验室，并已通过ISO17034、 药物杂质标准品联系人：黄舒琦 ISO9001体系认证。CATO中国95%以上员工均为本科以上学历，具有高 电话：020-81215950 度专业的化工化学行业经验。 非药物标准品联系人：叶淑明 目前为止，CATO中国已服务超1万家客户，提供的标准品覆盖工业 电话：020-81960175 品、食品、农药残留、兽药残留、环境、药物杂质、天然提取物等领域， E-mail：[email protected] 同时深度支持定制合成。 “扫一扫” 地址：广州市荔湾区西增路63号自编E1-A101A 我们相信，CATO能给您更好的产品及服务！ 关注CATO微信公众号 官网：http://www.cato-chem.com No. 货号 中文名 英文名 CAS号 药物杂质标准品 1 C3D-1016 2-氨基苯酚 2-Aminophenol 95-55-6 2 C3D-1331 门冬氨酸缩合物 (S)-2-((S)-3-Amino-3-carboxypropanamido)succinic acid 60079-22-3 3 C3D-1350 4-氯苯甲酸 4-Chlorobenzoic acid 74-11-3 4 C3D-1447 戊乙奎醚杂质4 Penehyclidine Impurity 4 5422-88-8 5 C3D-1481 氮卓斯汀EP杂质A Azelastine EP Impurity A 613-94-5 6 C3D-1593 氟西汀EP杂质A(盐酸托莫西汀相关化合物A) Fluoxetine EP Impurity A(Atomoxetine Related Compound A) 42142-52-9 7 C3D-1598 瑞巴派特 Rebamipide 90098-04-7 8 C3D-1636 4-甲基苯磺酸异丙酯 Isopropyl 4-Methylbenzenesulfonate 2307-69-9 9 C3D-1724 Methyl (R)-2-amino-2-phenylacetate hydrochloride Methyl (R)-2-Amino-2-Phenylacetate Hydrochloride 19883-41-1 10 C3D-1792 睾酮 Testosterone 58-22-0 11 C3D-1804 1-羟基苯并三唑 1-Hydroxybenzotriazole 2592-95-2 12 C3D-1808 苯甲醇 Benzyl alcohol 100-51-6 13 C3D-1823 2- [4-(溴甲基)苯基]丙酸 2-[4-(Bromomethyl)phenyl]propionic Acid 111128-12-2 14 C3D-1824 甲基-2-氧代环戊烷羧酸 Methyl 2-Oxocyclopentanecarboxylate 10472-24-9 15 C3D-1831 N-(2-甲基-5-硝基苯基)-4-(吡啶-3-基)嘧啶-2-胺 N-(2-Methyl-5-nitrophenyl)-4-(pyridin-3-yl)pyrimidin-2-amine 152460-09-8 16 C3D-1837 L-丙氨酰-L-谷氨酰胺 L-Alanyl-L-glutamine 39537-23-0 17 C3D-1848 1咪唑乙酸