https://m2.mtmt.hu/ hun 2026-09-22 07:55 JournalArticle 31236413 APPROVED true Institute of Enzymology, Research Centre for Natural Sciences of the Hungarian Academy of Sciences, Budapest, H-1117, Hungary Institute of Biochemistry, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, H-6726, Hungary Doctoral School of Multidisciplinary Medical Science, University of Szeged, Szeged, H-6720, Hungary School of Ph.D. Studies, Semmelweis University, Budapest, H-1085, Hungary Gene Design Ltd, Szeged, H-6726, Hungary Biospiral-2006 Ltd, Szeged, H-6726, Hungary Cited By :4 Export Date: 26 January 2022 Correspondence Address: Kulcsár, P.I.; Institute of Enzymology, Hungary; email: kulcsar.peter.istvan@ttk.mta.hu 0 false 2026-09-11T13:31:51.786+0000 2025-02-25T21:17:10.234+0000 2020-03-10T09:56:25.911+0000 true 10038471 Kulcsár Péter István /api/author/10038471 Author Kulcsár Péter István (Molekuláris biológia) true 10038471 2025-02-25T21:17:10.538+0000 2025-02-25T21:17:10.538+0000 true false true 24 24 /api/publicationtype/24 PublicationType Folyóiratcikk 1 true 24 JournalArticle true 10000059 Article true 24 24 /api/publicationtype/24 PublicationType Folyóiratcikk 1 true 24 JournalArticle /api/subtype/10000059 Szakcikk SubType Szakcikk (Folyóiratcikk) 101 true 10000059 true 1 /api/category/1 Category Tudományos true 1 Kulcsár, Péter István true true /api/journal/10015678 REVIEWED NATURE COMMUNICATIONS 2041-1723 true false 10015678 false 10015678 true 2041-1723 Journal FOREIGN 11 1 1223 1223 14 2020 Increased fidelity mutants of the SpCas9 nuclease constitute the most promising approach to mitigating its off-target effects. However, these variants are effective only in a restricted target space, and many of them are reported to work less efficiently when applied in clinically relevant, pre-assembled, ribonucleoprotein forms. The low tolerance to 5′-extended, 21G-sgRNAs contributes, to a great extent, to their decreased performance. Here, we report the generation of Blackjack SpCas9 variant that shows increased fidelity yet remain effective with 21G-sgRNAs. Introducing Blackjack mutations into previously reported increased fidelity variants make them effective with 21G-sgRNAs and increases their fidelity. Two “Blackjack” nucleases, eSpCas9<jats:italic>-plus</jats:italic> and SpCas9-HF1<jats:italic>-plus</jats:italic> are superior variants of eSpCas9 and SpCas9-HF1, respectively, possessing matching on-target activity and fidelity but retaining activity with 21G-sgRNAs. They facilitate the use of existing pooled sgRNA libraries with higher specificity and show similar activities whether delivered as plasmids or as pre-assembled ribonucleoproteins. true 2020 true true true false true false GOLD 2026-09-11 true https://doi.org/10.1038/s41467-020-15021-5 40 0 40 33 6 40 40 39 39 40 40 33 33 33 7 0 40 33 40 33 0 3 3 Folyóiratcikk Journal Article 24 39 Könyvrészlet Chapter in Book 25 1 Könyv Book 23 0 Egyéb konferenciaközlemény Conference paper 31 0 Egyéb konferenciakötet Conference proceedings 32 0 Oltalmi formák Protection forms 26 0 Disszertáció Thesis 28 0 Egyéb Miscellaneous 29 0 Alkotás Achievement 22 0 Kutatási adat Research data 33 0 Folyóiratcikk Journal Article 24 0 0 0 Könyvrészlet Chapter in Book 25 0 0 0 Könyv Book 23 0 0 0 Egyéb konferenciaközlemény Conference paper 31 0 0 0 Egyéb konferenciakötet Conference proceedings 32 0 0 0 Oltalmi formák Protection forms 26 0 0 0 Disszertáció Thesis 28 0 0 0 Egyéb Miscellaneous 29 0 0 0 Alkotás Achievement 22 0 0 0 Kutatási adat Research data 33 0 0 0 2020 0 2 2 2 2 2021 0 2 0 2 0 2022 0 10 8 10 8 2023 0 9 8 9 8 2024 0 7 7 7 7 2025 0 5 5 5 5 2026 0 5 3 5 3 D1 A nemzetközi false 10020968,10001345,10055848,10038471 false false false 2020-06-08T08:56:26.776+0000 Biokémiai Intézet (HRN SZBK); Doktori Iskola (SE); Enzimológiai Intézet (TTK); Multidiszciplináris Orvostudományok Doktori Iskola (SZTE / DI) 6 30 4 7 0 true Language 10002 /api/language/10002 Angol Angol English true 2 true PersonAuthorship 91042177 /api/authorship/91042177 Kulcsár, Péter István ✉ [Kulcsár, Péter István (Molekuláris biológia), szerző] Biokémiai Intézet (HRN SZBK); Multidiszciplináris Orvostudományok Doktori Iskola (SZTE / DI) 1 0.14285715 true false true Author 10038471 /api/author/10038471 Kulcsár Péter István (Molekuláris biológia) Kulcsár Péter István true 10038471 true Kulcsár Péter István true false false AuthorshipType 1 /api/authorshiptype/1 Szerző 0 true 0 true false true PersonAuthorship 91042178 /api/authorship/91042178 Tálas, András [Tálas, András (Molekuláris Biológus), szerző] Enzimológiai Intézet (TTK); Doktori Iskola (SE) 2 0.14285715 false false false Author 10055848 /api/author/10055848 Tálas András (Molekuláris Biológus) Tálas András true 10055848 true Tálas András true false false AuthorshipType 1 /api/authorshiptype/1 Szerző 0 true 0 true false true PersonAuthorship 91042179 /api/authorship/91042179 Tóth, Eszter [Tóth, Eszter (molekuláris biológia), szerző] 3 0.14285715 false false false Author 10020968 /api/author/10020968 Tóth Eszter (molekuláris biológia) Tóth Eszter true 10020968 true Tóth Eszter true false false AuthorshipType 1 /api/authorshiptype/1 Szerző 0 true 0 true false true PersonAuthorship 91042180 /api/authorship/91042180 Nyeste, Antal [Nyeste, Antal (Biológus), szerző] 4 0.14285715 false false false Author 10039251 /api/author/10039251 Nyeste Antal (Biológus) Nyeste Antal true 10039251 true Nyeste Antal true false false AuthorshipType 1 /api/authorshiptype/1 Szerző 0 true 0 true false true PersonAuthorship 91042181 /api/authorship/91042181 Ligeti, Zoltán [Ligeti, Zoltán Mihály (Biológia), szerző] 5 0.14285715 false false false Author 10069721 /api/author/10069721 Ligeti Zoltán Mihály (Biológia) Ligeti Zoltán Mihály true true Ligeti Zoltán true false false AuthorshipType 1 /api/authorshiptype/1 Szerző 0 true 0 true false true PersonAuthorship 91042182 /api/authorship/91042182 Welker, Zsombor 6 0.14285715 false false false Welker Zsombor true false false AuthorshipType 1 /api/authorshiptype/1 Szerző 0 true 0 true false true PersonAuthorship 91042183 /api/authorship/91042183 Welker, Ervin [Welker, Ervin (Fehérje feltekeredés), szerző] Biokémiai Intézet (HRN SZBK); Enzimológiai Intézet (TTK) 7 0.14285715 false true false Author 10001345 /api/author/10001345 Welker Ervin (Fehérje feltekeredés) Welker Ervin true 10001345 true Welker Ervin true false false AuthorshipType 1 /api/authorshiptype/1 Szerző 0 true 0 true false true PublicationIdentifier 16745496 /api/publicationidentifier/16745496 DOI: 10.1038/s41467-020-15021-5 PlainSource 6 /api/publicationsource/6 DOI PublicationSourceType 10001 /api/publicationsourcetype/10001 DOI true true true DOI DOI https://doi.org/@@@ true true 6 true GOLD true IDENTICAL 10.1038/s41467-020-15021-5 https://doi.org/10.1038/s41467-020-15021-5 false true PublicationIdentifier 16755217 /api/publicationidentifier/16755217 REAL: 107176 SwordSource 36 /api/publicationsource/36 REAL PublicationSourceType 10007 /api/publicationsourcetype/10007 Repozitórium true true true REAL REAL https://real.mtak.hu/@@@ true true 36 true GREEN true 107176 https://real.mtak.hu/107176 false true PublicationIdentifier 17078832 /api/publicationidentifier/17078832 WoS: 000546777300001 PlainSource 1 /api/publicationsource/1 WoS PublicationSourceType 10003 /api/publicationsourcetype/10003 Indexelő adatbázis false true true WoS WoS https://www.webofscience.com/wos/woscc/full-record/@@@ true true 1 true IDENTICAL 000546777300001 https://www.webofscience.com/wos/woscc/full-record/000546777300001 false true PublicationIdentifier 17047209 /api/publicationidentifier/17047209 Scopus: 85081530340 PlainSource 3 /api/publicationsource/3 Scopus PublicationSourceType 10003 /api/publicationsourcetype/10003 Indexelő adatbázis false true true Scopus Scopus http://www.scopus.com/record/display.url?origin=inward&eid=2-s2.0-@@@ true true 3 true IDENTICAL 85081530340 http://www.scopus.com/record/display.url?origin=inward&eid=2-s2.0-85081530340 false true PublicationIdentifier 16754993 /api/publicationidentifier/16754993 PubMed: 32144253 PlainSource 17 /api/publicationsource/17 PubMed PublicationSourceType 10003 /api/publicationsourcetype/10003 Indexelő adatbázis false true true PubMed PubMed http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=@@@&dopt=Abstract true true 17 true IDENTICAL 32144253 http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=32144253&dopt=Abstract false true MtaRating 11551153 /api/mtarating/11551153 IX. Regionális Tudományok Bizottsága:A nemzetközi NATURE COMMUNICATIONS 2041-1723 RatingType 10010 /api/ratingtype/10010 IX. Regionális Tudományok Bizottsága RegTB Institute 14100 /api/institute/14100 Regionális Tudományok Bizottsága IXGJO RTB [1901-] 0 false true false Regionális Tudományok Bizottsága Committee on Regional Studies true 14100 true true true A nemzetközi false true SjrRating 10751794 /api/sjrrating/10751794 sjr:D1 (2020) Scopus - Biochemistry, Genetics and Molecular Biology (miscellaneous) NATURE COMMUNICATIONS 2041-1723 11 0.1 journal RatingType 10002 /api/ratingtype/10002 sjr sjr true true ClassificationExternal 1301 /api/classificationexternal/1301 Scopus - Biochemistry, Genetics and Molecular Biology (miscellaneous) true 1301 true D1 DIRECT true true Reference 2604402 /api/reference/2604402 1. Jinek, M. et al. A programmable dual-RNA-guided DNA endonuclease in adaptive bacterial immunity. Science 337, 816–821 (2012)., DOI: 10.1126/science.1225829 1 10.1126/science.1225829 false true Reference 2604403 /api/reference/2604403 2. Mali, P., Esvelt, K. M. & Church, G. M. Cas9 as a versatile tool for engineering biology. Nat. Methods 10, 957–963 (2013)., DOI: 10.1038/nmeth.2649 2 10.1038/nmeth.2649 false true Reference 2604404 /api/reference/2604404 3. Jinek, M. et al. RNA-programmed genome editing in human cells. elife 2, e00471 (2013)., DOI: 10.7554/eLife.00471 3 10.7554/eLife.00471 false true Reference 2604405 /api/reference/2604405 4. Mali, P. et al. RNA-guided human genome engineering via Cas9. Science 339, 823–826 (2013)., DOI: 10.1126/science.1232033 4 10.1126/science.1232033 false true Reference 2604406 /api/reference/2604406 5. Cong, L. et al. Multiplex genome engineering using CRISPR/Cas systems. Science 339, 819–823 (2013)., DOI: 10.1126/science.1231143 5 10.1126/science.1231143 false true Reference 2604407 /api/reference/2604407 6. Cho, S. W., Kim, S., Kim, J. M. & Kim, J. S. Targeted genome engineering in human cells with the Cas9 RNA-guided endonuclease. Nat. Biotechnol. 31, 230–232 (2013)., DOI: 10.1038/nbt.2507 6 10.1038/nbt.2507 false true Reference 2604408 /api/reference/2604408 7. Hsu, P. D., Lander, E. S. & Zhang, F. Development and applications of CRISPR-Cas9 for genome engineering. Cell 157, 1262–1278 (2014)., DOI: 10.1016/j.cell.2014.05.010 7 10.1016/j.cell.2014.05.010 false true Reference 2604409 /api/reference/2604409 8. Sander, J. D. & Joung, J. K. CRISPR-Cas systems for editing, regulating and targeting genomes. Nat. Biotechnol. 32, 347–355 (2014)., DOI: 10.1038/nbt.2842 8 10.1038/nbt.2842 false true Reference 2604410 /api/reference/2604410 9. Tanenbaum, M. E., Gilbert, L. A., Qi, L. S., Weissman, J. S. & Vale, R. D. A protein-tagging system for signal amplification in gene expression and fluorescence imaging. Cell 159, 635–646 (2014)., DOI: 10.1016/j.cell.2014.09.039 9 10.1016/j.cell.2014.09.039 false true Reference 2604411 /api/reference/2604411 10. Zuris, J. A. et al. Cationic lipid-mediated delivery of proteins enables efficient protein-based genome editing in vitro and in vivo. Nat. Biotechnol. 33, 73–80 (2015)., DOI: 10.1038/nbt.3081 10 10.1038/nbt.3081 false true Reference 2604412 /api/reference/2604412 11. Vora, S., Tuttle, M., Cheng, J. & Church, G. Next stop for the CRISPR revolution: RNA-guided epigenetic regulators. FEBS J. 283, 3181–3193 (2016)., DOI: 10.1111/febs.13768 11 10.1111/febs.13768 false true Reference 2604413 /api/reference/2604413 12. Koonin, E. V., Makarova, K. S. & Zhang, F. Diversity, classification and evolution of CRISPR-Cas systems. Curr. Opin. Microbiol. 37, 67–78 (2017)., DOI: 10.1016/j.mib.2017.05.008 12 10.1016/j.mib.2017.05.008 false true Reference 2604414 /api/reference/2604414 13. Komor, A. C., Badran, A. H. & Liu, D. R. CRISPR-based technologies for the manipulation of eukaryotic genomes. Cell 169, 559 (2017)., DOI: 10.1016/j.cell.2017.04.005 13 10.1016/j.cell.2017.04.005 false true Reference 2604415 /api/reference/2604415 14. Wu, W. Y., Lebbink, J. H. G., Kanaar, R., Geijsen, N. & van der Oost, J. Genome editing by natural and engineered CRISPR-associated nucleases. Nat. Chem. Biol. 14, 642–651 (2018)., DOI: 10.1038/s41589-018-0080-x 14 10.1038/s41589-018-0080-x false true Reference 2604416 /api/reference/2604416 15. Mali, P. et al. CAS9 transcriptional activators for target specificity screening and paired nickases for cooperative genome engineering. Nat. Biotechnol. 31, 833–838 (2013)., DOI: 10.1038/nbt.2675 15 10.1038/nbt.2675 false true Reference 2604417 /api/reference/2604417 16. Guilinger, J. P., Thompson, D. B. & Liu, D. R. Fusion of catalytically inactive Cas9 to FokI nuclease improves the specificity of genome modification. Nat. Biotechnol. 32, 577–582 (2014)., DOI: 10.1038/nbt.2909 16 10.1038/nbt.2909 false true Reference 2604418 /api/reference/2604418 17. Tsai, S. Q. et al. Dimeric CRISPR RNA-guided FokI nucleases for highly specific genome editing. Nat. Biotechnol. 32, 569–576 (2014)., DOI: 10.1038/nbt.2908 17 10.1038/nbt.2908 false true Reference 2604419 /api/reference/2604419 18. Wyvekens, N., Topkar, V. V., Khayter, C., Joung, J. K. & Tsai, S. Q. Dimeric CRISPR RNA-guided FokI-dCas9 nucleases directed by truncated gRNAs for highly specific genome editing. Hum. Gene Ther. 26, 425–431 (2015)., DOI: 10.1089/hum.2015.084 18 10.1089/hum.2015.084 false true Reference 2604420 /api/reference/2604420 19. Ran, F. A. et al. Double nicking by RNA-guided CRISPR Cas9 for enhanced genome editing specificity. Cell 154, 1380–1389 (2013)., DOI: 10.1016/j.cell.2013.08.021 19 10.1016/j.cell.2013.08.021 false true Reference 2604421 /api/reference/2604421 20. Cho, S. W. et al. Analysis of off-target effects of CRISPR/Cas-derived RNA-guided endonucleases and nickases. Genome Res 24, 132–141 (2014)., DOI: 10.1101/gr.162339.113 20 10.1101/gr.162339.113 false true Reference 2604422 /api/reference/2604422 21. Fu, Y., Sander, J. D., Reyon, D., Cascio, V. M. & Joung, J. K. Improving CRISPR-Cas nuclease specificity using truncated guide RNAs. Nat. Biotechnol. 32, 279–284 (2014)., DOI: 10.1038/nbt.2808 21 10.1038/nbt.2808 false true Reference 2604423 /api/reference/2604423 22. Fu, Y., Reyon, D. & Joung, J. K. Targeted genome editing in human cells using CRISPR/Cas nucleases and truncated guide RNAs. Methods Enzymol. 546, 21–45 (2014)., DOI: 10.1016/B978-0-12-801185-0.00002-7 22 10.1016/B978-0-12-801185-0.00002-7 false true Reference 2604424 /api/reference/2604424 23. Tsai, S. Q. et al. GUIDE-seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases. Nat. Biotechnol. 33, 187–197 (2015)., DOI: 10.1038/nbt.3117 23 10.1038/nbt.3117 false true Reference 2604425 /api/reference/2604425 24. Slaymaker, I. M. et al. Rationally engineered Cas9 nucleases with improved specificity. Science 351, 84–88 (2016)., DOI: 10.1126/science.aad5227 24 10.1126/science.aad5227 false true Reference 2604426 /api/reference/2604426 25. Kleinstiver, B. P. et al. High-fidelity CRISPR-Cas9 nucleases with no detectable genome-wide off-target effects. Nature 529, 490–495 (2016)., DOI: 10.1038/nature16526 25 10.1038/nature16526 false true Reference 2604427 /api/reference/2604427 26. Chen, J. S. et al. Enhanced proofreading governs CRISPR-Cas9 targeting accuracy. Nature 550, 407–410 (2017)., DOI: 10.1038/nature24268 26 10.1038/nature24268 false true Reference 2604428 /api/reference/2604428 27. Casini, A. et al. A highly specific SpCas9 variant is identified by in vivo screening in yeast. Nat. Biotechnol. 36, 265–271 (2018)., DOI: 10.1038/nbt.4066 27 10.1038/nbt.4066 false true Reference 2604429 /api/reference/2604429 28. Kulcsar, P. I. et al. Crossing enhanced and high fidelity SpCas9 nucleases to optimize specificity and cleavage. Genome Biol. 18, 190 (2017)., DOI: 10.1186/s13059-017-1318-8 28 10.1186/s13059-017-1318-8 false true Reference 2604430 /api/reference/2604430 29. Kim, S., Bae, T., Hwang, J. & Kim, J. S. Rescue of high-specificity Cas9 variants using sgRNAs with matched 5′ nucleotides. Genome Biol. 18, 218 (2017)., DOI: 10.1186/s13059-017-1355-3 29 10.1186/s13059-017-1355-3 false true Reference 2604431 /api/reference/2604431 30. Zhang, D. et al. Perfectly matched 20-nucleotide guide RNA sequences enable robust genome editing using high-fidelity SpCas9 nucleases. Genome Biol. 18, 191 (2017)., DOI: 10.1186/s13059-017-1325-9 30 10.1186/s13059-017-1325-9 false true Reference 2604432 /api/reference/2604432 31. Anders, C., Niewoehner, O., Duerst, A. & Jinek, M. Structural basis of PAM-dependent target DNA recognition by the Cas9 endonuclease. Nature 513, 569–573 (2014)., DOI: 10.1038/nature13579 31 10.1038/nature13579 false true Reference 2604433 /api/reference/2604433 32. Jinek, M. et al. Structures of Cas9 endonucleases reveal RNA-mediated conformational activation. Science 343, 1247997 (2014)., DOI: 10.1126/science.1247997 32 10.1126/science.1247997 false true Reference 2604434 /api/reference/2604434 33. Kocak, D.D. et al. Increasing the specificity of CRISPR systems with engineered RNA secondary structures. Nat. Biotechnol. 37, 657–666 (2019)., DOI: 10.1038/s41587-019-0095-1 33 10.1038/s41587-019-0095-1 false true Reference 2604435 /api/reference/2604435 34. Goomer, R. S. & Kunkel, G. R. The transcriptional start site for a human U6 small nuclear RNA gene is dictated by a compound promoter element consisting of the PSE and the TATA box. Nucleic Acids Res. 20, 4903–4912 (1992)., DOI: 10.1093/nar/20.18.4903 34 10.1093/nar/20.18.4903 false true Reference 2604436 /api/reference/2604436 35. Milligan, J. F., Groebe, D. R., Witherell, G. W. & Uhlenbeck, O. C. Oligoribonucleotide synthesis using T7 RNA polymerase and synthetic DNA templates. Nucleic Acids Res. 15, 8783–8798 (1987)., DOI: 10.1093/nar/15.21.8783 35 10.1093/nar/15.21.8783 false true Reference 2604437 /api/reference/2604437 36. Beckert, B. & Masquida, B. Synthesis of RNA by in vitro transcription. Methods Mol. Biol. 703, 29–41 (2011)., DOI: 10.1007/978-1-59745-248-9_3 36 10.1007/978-1-59745-248-9_3 false true Reference 2604438 /api/reference/2604438 37. Moreno-Mateos, M. A. et al. CRISPRscan: designing highly efficient sgRNAs for CRISPR-Cas9 targeting in vivo. Nat. Methods 12, 982–988 (2015)., DOI: 10.1038/nmeth.3543 37 10.1038/nmeth.3543 false true Reference 2604439 /api/reference/2604439 38. Nowak, C. M., Lawson, S., Zerez, M., Bleris, L. & Guide, R. N. A. engineering for versatile Cas9 functionality. Nucleic Acids Res 44, 9555–9564 (2016). 38 false true Reference 2604440 /api/reference/2604440 39. Gao, Y. & Zhao, Y. Self-processing of ribozyme-flanked RNAs into guide RNAs in vitro and in vivo for CRISPR-mediated genome editing. J. Integr. Plant Biol. 56, 343–349 (2014)., DOI: 10.1111/jipb.12152 39 10.1111/jipb.12152 false true Reference 2604441 /api/reference/2604441 40. Nissim, L., Perli, S. D., Fridkin, A., Perez-Pinera, P. & Lu, T. K. Multiplexed and programmable regulation of gene networks with an integrated RNA and CRISPR/Cas toolkit in human cells. Mol. Cell 54, 698–710 (2014)., DOI: 10.1016/j.molcel.2014.04.022 40 10.1016/j.molcel.2014.04.022 false true Reference 2604442 /api/reference/2604442 41. Xie, K., Minkenberg, B. & Yang, Y. Boosting CRISPR/Cas9 multiplex editing capability with the endogenous tRNA-processing system. Proc. Natl Acad. Sci. USA 112, 3570–3575 (2015)., DOI: 10.1073/pnas.1420294112 41 10.1073/pnas.1420294112 false true Reference 2604443 /api/reference/2604443 42. Vakulskas, C. A. et al. A high-fidelity Cas9 mutant delivered as a ribonucleoprotein complex enables efficient gene editing in human hematopoietic stem and progenitor cells. Nat. Med. 24, 1216–1224 (2018)., DOI: 10.1038/s41591-018-0137-0 42 10.1038/s41591-018-0137-0 false true Reference 2604444 /api/reference/2604444 43. Jiang, F. et al. Structures of a CRISPR-Cas9 R-loop complex primed for DNA cleavage. Science 351, 867–871 (2016)., DOI: 10.1126/science.aad8282 43 10.1126/science.aad8282 false true Reference 2604445 /api/reference/2604445 44. Nishimasu, H. et al. Crystal structure of Cas9 in complex with guide RNA and target DNA. Cell 156, 935–949 (2014)., DOI: 10.1016/j.cell.2014.02.001 44 10.1016/j.cell.2014.02.001 false true Reference 2604446 /api/reference/2604446 45. Lee, J. K. et al. Directed evolution of CRISPR-Cas9 to increase its specificity. Nat. Commun. 9, 3048 (2018)., DOI: 10.1038/s41467-018-05477-x 45 10.1038/s41467-018-05477-x false true Reference 2604447 /api/reference/2604447 46. Dong, F., Xie, K., Chen, Y., Yang, Y. & Mao, Y. Polycistronic tRNA and CRISPR guide-RNA enables highly efficient multiplexed genome engineering in human cells. Biochem. Biophys. Res. Commun. 482, 889–895 (2017)., DOI: 10.1016/j.bbrc.2016.11.129 46 10.1016/j.bbrc.2016.11.129 false true Reference 2604448 /api/reference/2604448 47. Lee, R. T., Ng, A. S. & Ingham, P. W. Ribozyme mediated gRNA generation for in vitro and in vivo CRISPR/Cas9 mutagenesis. PLoS ONE 11, e0166020 (2016)., DOI: 10.1371/journal.pone.0166020 47 10.1371/journal.pone.0166020 false true Reference 2604449 /api/reference/2604449 48. Talas, A. et al. A convenient method to pre-screen candidate guide RNAs for CRISPR/Cas9 gene editing by NHEJ-mediated integration of a ‘self-cleaving’ GFP-expression plasmid. DNA Res. 24, 609–621 (2017)., DOI: 10.1093/dnares/dsx029 48 10.1093/dnares/dsx029 false true Reference 2604450 /api/reference/2604450 49. Nishimasu, H. et al. Engineered CRISPR-Cas9 nuclease with expanded targeting space. Science 361, 1259–1262 (2018)., DOI: 10.1126/science.aas9129 49 10.1126/science.aas9129 false true Reference 2604451 /api/reference/2604451 50. Hu, J. H. et al. Evolved Cas9 variants with broad PAM compatibility and high DNA specificity. Nature 556, 57–63 (2018)., DOI: 10.1038/nature26155 50 10.1038/nature26155 false true Reference 2604452 /api/reference/2604452 51. Engler, C., Kandzia, R. & Marillonnet, S. A one pot, one step, precision cloning method with high throughput capability. PloS ONE 3, e3647 (2008)., DOI: 10.1371/journal.pone.0003647 51 10.1371/journal.pone.0003647 false true Reference 2604453 /api/reference/2604453 52. Kostylev, M., Otwell, A.E., Richardson, R.E. & Suzuki, Y. Cloning should be simple: Escherichia coli DH5 alpha-mediated assembly of multiple DNA fragments with short end homologies. PLoS ONE 10, e0137466 (2015)., DOI: 10.1371/journal.pone.0137466 52 10.1371/journal.pone.0137466 false true Reference 2604454 /api/reference/2604454 53. Tóth, E. et al. Restriction enzyme body doubles and PCR cloning: on the general use of type IIs restriction enzymes for cloning. PloS ONE 9, e90896 (2014)., DOI: 10.1371/journal.pone.0090896 53 10.1371/journal.pone.0090896 false true Reference 2604455 /api/reference/2604455 54. Konermann, S. et al. Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature 517, 583–588 (2015)., DOI: 10.1038/nature14136 54 10.1038/nature14136 false true Reference 2604456 /api/reference/2604456 55. Iwamoto, M., Bjorklund, T., Lundberg, C., Kirik, D. & Wandless, T. J. A general chemical method to regulate protein stability in the mammalian central nervous system. Chem. Biol. 17, 981–988 (2010)., DOI: 10.1016/j.chembiol.2010.07.009 55 10.1016/j.chembiol.2010.07.009 false true Reference 2604457 /api/reference/2604457 56. Sanjana, N. E., Shalem, O. & Zhang, F. Improved vectors and genome-wide libraries for CRISPR screening. Nat. Methods 11, 783–784 (2014)., DOI: 10.1038/nmeth.3047 56 10.1038/nmeth.3047 false true Reference 2604458 /api/reference/2604458 57. Vriend, L. E., Jasin, M. & Krawczyk, P. M. Assaying break and nick-induced homologous recombination in mammalian cells using the DR-GFP reporter and Cas9 nucleases. Methods Enzymol. 546, 175–191 (2014)., DOI: 10.1016/B978-0-12-801185-0.00009-X 57 10.1016/B978-0-12-801185-0.00009-X false true Reference 2604459 /api/reference/2604459 58. Tsai, S. Q., Topkar, V. V., Joung, J. K. & Aryee, M. J. Open-source guideseq software for analysis of GUIDE-seq data. Nat. Biotechnol. 34, 483 (2016)., DOI: 10.1038/nbt.3534 58 10.1038/nbt.3534 false true Reference 2604460 /api/reference/2604460 59. Brinkman, E. K., Chen, T., Amendola, M. & van Steensel, B. Easy quantitative assessment of genome editing by sequence trace decomposition. Nucleic Acids Res. 42, e168 (2014)., DOI: 10.1093/nar/gku936 59 10.1093/nar/gku936 false true Reference 2604461 /api/reference/2604461 60. Spitzer, M., Wildenhain, J., Rappsilber, J. & Tyers, M. BoxPlotR: a web tool for generation of box plots. Nat. Methods 11, 121–122 (2014)., DOI: 10.1038/nmeth.2811 60 10.1038/nmeth.2811 false true /api/publication/31236413 Kulcsár Péter István et al. Blackjack mutations improve the on-target activities of increased fidelity variants of SpCas9 with 5′G-extended sgRNAs. (2020) NATURE COMMUNICATIONS 2041-1723 11 1<div class="JournalArticle Publication long-list"> <div class="authors"> <img title="Forrásközlemény" style="float: left" src="/frontend/resources/grid/publication-core-icon.png"> <img title="Idézőközlemény" style="float: left" src="/frontend/resources/grid/publication-citation-icon.png"> <div class="autype autype0"> <span class="author-name" mtid="10038471"><a href="/gui2/?type=authors&mode=browse&sel=10038471" target="_blank">Kulcsár Péter István ✉ (<span class="authorship-author-name">Kulcsár Péter István</span> <span class="authorAux-mtmt"> Molekuláris biológia</span>) </a> </span> <span class="author-affil"><span title="Szegedi Biológiai Kutatóközpont">SZBK</span>/Biokémiai Intézet; <span title="Szegedi Tudományegyetem">SZTE</span>/<span title="Doktori Iskolák">DI</span>/Multidiszciplináris Orvostudományok Doktori Iskola</span> ;&nbsp;&nbsp;&nbsp; <span class="author-name" mtid="10055848"><a href="/gui2/?type=authors&mode=browse&sel=10055848" target="_blank">Tálas András (<span class="authorship-author-name">Tálas András</span> <span class="authorAux-mtmt"> Molekuláris Biológus</span>) </a> </span> <span class="author-affil"><span title="Természettudományi Kutatóközpont">TTK</span>/Enzimológiai Intézet; <span title="Semmelweis Egyetem">SE</span>/Doktori Iskola</span> ;&nbsp;&nbsp;&nbsp; <span class="author-name" mtid="10020968"><a href="/gui2/?type=authors&mode=browse&sel=10020968" target="_blank">Tóth Eszter (<span class="authorship-author-name">Tóth Eszter</span> <span class="authorAux-mtmt"> molekuláris biológia</span>) </a> </span> ;&nbsp;&nbsp;&nbsp; <span class="author-name" mtid="10039251"><a href="/gui2/?type=authors&mode=browse&sel=10039251" target="_blank">Nyeste Antal (<span class="authorship-author-name">Nyeste Antal</span> <span class="authorAux-mtmt"> Biológus</span>) </a> </span> ;&nbsp;&nbsp;&nbsp; <span class="author-name" mtid="10069721"><a href="/gui2/?type=authors&mode=browse&sel=10069721" target="_blank">Ligeti Zoltán (<span class="authorship-author-name">Ligeti Zoltán Mihály</span> <span class="authorAux-mtmt"> Biológia</span>) </a> </span> ;&nbsp;&nbsp;&nbsp; <span class="author-name" >Welker Zsombor </span> ;&nbsp;&nbsp;&nbsp; <span class="author-name" mtid="10001345"><a href="/gui2/?type=authors&mode=browse&sel=10001345" target="_blank">Welker Ervin (<span class="authorship-author-name">Welker Ervin</span> <span class="authorAux-mtmt"> Fehérje feltekeredés</span>) </a> </span> <span class="author-affil"><span title="Szegedi Biológiai Kutatóközpont">SZBK</span>/Biokémiai Intézet; <span title="Természettudományi Kutatóközpont">TTK</span>/Enzimológiai Intézet</span> </div> </div> <div class="title"><a href="/gui2/?mode=browse&params=publication;31236413" target="_blank">Blackjack mutations improve the on-target activities of increased fidelity variants of SpCas9 with 5′G-extended sgRNAs</a></div> <div> <span class="journal-title">NATURE COMMUNICATIONS</span> <span class="journal-issn">( <a target="_blank" href="https://portal.issn.org/resource/ISSN/2041-1723">2041-1723</a>)</span>: <span class="journal-volume">11</span> <span class="journal-issue">1</span> <span class="page"> Paper 1223. 14 p. </span> <span class="year">(2020)</span> </div> <div class="pub-footer"> <span class="language" xmlns="http://www.w3.org/1999/html">Nyelv: Angol | </span> <span class="identifiers"> <span class="id identifier oa_GOLD" title=" Gold "> <a style="color:blue" title="10.1038/s41467-020-15021-5" target="_blank" href="https://doi.org/10.1038/s41467-020-15021-5"> DOI </a> </span> <span class="id identifier oa_GREEN" title=" Green "> <a style="color:black" title="107176" target="_blank" href="https://real.mtak.hu/107176"> REAL </a> </span> <span class="id identifier oa_none" title="none"> <a style="color:blue" title="000546777300001" target="_blank" href="https://www.webofscience.com/wos/woscc/full-record/000546777300001"> WoS </a> </span> <span class="id identifier oa_none" title="none"> <a style="color:blue" title="85081530340" target="_blank" href="http://www.scopus.com/record/display.url?origin=inward&eid=2-s2.0-85081530340"> Scopus </a> </span> <span class="id identifier oa_none" title="none"> <a style="color:blue" title="32144253" target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=32144253&dopt=Abstract"> PubMed </a> </span> </span> <OnlyViewableByAuthor><div class="ratings"> <div class="journal-subject">Folyóirat szakterülete: Scopus - Biochemistry, Genetics and Molecular Biology (miscellaneous)&nbsp;&nbsp;&nbsp;SJR indikátor:&nbsp;D1</div> <div class="journal-subject">Folyóirat szakterülete: Scopus - Chemistry (miscellaneous)&nbsp;&nbsp;&nbsp;SJR indikátor:&nbsp;D1</div> <div class="journal-subject">Folyóirat szakterülete: Scopus - Physics and Astronomy (miscellaneous)&nbsp;&nbsp;&nbsp;SJR indikátor:&nbsp;D1</div> <div class="journal-subject">Regionális Tudományok Bizottsága IXGJO RTB [1901-]&nbsp;&nbsp;&nbsp;A nemzetközi</div> </div></OnlyViewableByAuthor> <div class="publication-citation" style="margin-left: 0.5cm;"> <span title="Nyilvános idézőközlemények összesen, említések nélkül" class="citingPub-count">Nyilvános idéző összesen: 40</span> | Független: 33 | Függő: 7 | Nem jelölt: 0 | WoS jelölt: 39 | Scopus jelölt:&nbsp;39 | WoS/Scopus jelölt:&nbsp;40 | DOI jelölt:&nbsp;40 </div> <div class="publication-citation"> <a target="_blank" href="/api/publication?cond=citations.related;eq;31236413&sort=publishedYear,desc&sort=title"> Idézett közlemények száma: 3 </a> </div> <div class="mtid"><span class="long-pub-mtid">Közlemény: 31236413</span> | <span class="status-data status-APPROVED"> Nyilvános </span> Forrás Idéző | <span class="type-subtype">Folyóiratcikk ( Szakcikk ) </span> | <span class="pub-category">Tudományos</span> | <span class="publication-sourceOfData">kézi felvitel</span> </div> <div class="lastModified">Utolsó módosítás: 2025.02.25. 22:17 Ladányi Gusztáv (MTMT API user, admin) </div> <pre class="comment" style="margin-top: 0; margin-bottom: 0;"><u>Megjegyzés</u>: Institute of Enzymology, Research Centre for Natural Sciences of the Hungarian Academy of Sciences, Budapest, H-1117, Hungary Institute of Biochemistry, Biological Research Centre of the Hungarian Academy of Sciences, Szeged, H-6726, Hungary Doctoral School of Multidisciplinary Medical Science, University of Szeged, Szeged, H-6720, Hungary ...</pre> </div></div>