Title:
ARGONAUTE PROTEIN MUTANT AND USE THEREOF
Document Type and Number:
WIPO Patent Application WO/2019/169945
Kind Code:
A1
Abstract:
The present invention relates to a mutant of Argonaute protein lacking DNA cleavage activity but having DNA binding activity, the mutation of the mutant being located in the PIWI domain. The present invention also relates to use based on the protein mutant, especially in enrichment of target DNA and construction of sequencing libraries. Therefore, the present invention also relates to a method for enrichment of a target DNA, comprising the following steps: (a) designing a guide sequence for a specific sequence in a target DNA; (b) binding the mutant, the guide sequence and the target DNA according to the present invention to obtain a mutant-guide sequence-target DNA ternary complex; (c) capturing the mutant-guide sequence-target DNA ternary complex by means of a capture medium; and (d) isolating the target DNA from the captured mutant-guide sequence-target DNA ternary complex to obtain an enriched target DNA.
Inventors:
ZHANG JIANGUANG (CN)
MAO AIPING (CN)
MAO AIPING (CN)
Application Number:
PCT/CN2019/070253
Publication Date:
September 12, 2019
Filing Date:
January 03, 2019
Export Citation:
Assignee:
BERRY GENOMICS CO LTD (CN)
International Classes:
C12N9/22; C40B50/06
Foreign References:
| CN108048532A | 2018-05-18 |
Other References:
WANG, Y.: "Nucleation, Propagation and Cleavage of Target RNAs in Ago Silencing Complexes", NATURE, vol. 461, no. 7265, 8 October 2009 (2009-10-08), XP055265388
See also references of EP 3763812A4
GARCIA-GARCIA, G ET AL.: "Assessment of the latest NGS enrichment capture methods in clinical context", SCI REP, vol. 6, 2016, pages 20948, XP055623863, DOI: 10.1038/srep20948
BODI, K. ET AL.: "Comparison of commercially available target enrichment methods for next-generation sequencing", J BIOMOL TECH, vol. 24, 2013, pages 73 - 86
NEWMAN, A. M. ET AL.: "Integrated digital error suppression for improved detection of circulating tumor DNA", NAT BIOTECHNOL, 2016
SAMORODNITSKY, E. ET AL.: "Evaluation of Hybridization Capture Versus Amplicon-Based Methods for Whole-Exome Sequencing", HUM MUTAT, vol. 36, 2015, pages 903 - 914
KUSCU, C.ARSLAN, S.SINGH, R.THORPE, J.ADLI, M.: "Genome-wide analysis reveals characteristics of off-target sites bound by the Cas9 endonuclease", NAT BIOTECHNOL, vol. 32, 2014, pages 677 - 683, XP055382577, DOI: 10.1038/nbt.2916
WU, X. ET AL.: "Genome-wide binding of the CRISPR endonuclease Cas9 in mammalian cells.", NAT BIOTECHNOL, vol. 32, 2014, pages 670 - 676, XP055241568, DOI: 10.1038/nbt.2889
LIU, X. ET AL.: "In Situ Capture of Chromatin Interactions by Biotinylated dCas9", CELL, vol. 170, 2017, pages 1028 - 1043
FUJITA, T.YUNO, M.FUJII, H.: "Efficient sequence-specific isolation of DNA fragments and chromatin by in vitro enChIP technology using recombinant CRISPR ribonucleoproteins", GENES CELLS, vol. 21, 2016, pages 370 - 377, XP055621734, DOI: 10.1111/gtc.12341
SWARTS, D. C. ET AL.: "The evolutionary journey of Argonaute proteins", NAT STRUCT MOL BIOL, vol. 21, 2014, pages 743 - 753, XP055287457, DOI: 10.1038/nsmb.2879
SONG, J. J.SMITH, S. K.HANNON, G. J.JOSHUA-TOR, L.: "Crystal structure of Argonaute and its implications for RISC slicer activity", SCIENCE, vol. 305, 2004, pages 1434 - 1437, XP002470480, DOI: 10.1126/science.1102514
SWARTS, D. C. ET AL.: "Argonaute of the archaeon Pyrococcus furiosus is a DNA-guided nuclease that targets cognate DNA", NUCLEIC ACIDS RES, vol. 43, 2015, pages 5120 - 5129, XP055287460, DOI: 10.1093/nar/gkv415
RAINES, R. T.MCCORMICK, M.VAN OOSBREE, T. R.MIERENDORF, R. C.: "The S.Tag fusion system for protein purification.", METHODS ENZYMOL, vol. 326, 2000, pages 362 - 376, XP009135533
See also references of EP 3763812A4
GARCIA-GARCIA, G ET AL.: "Assessment of the latest NGS enrichment capture methods in clinical context", SCI REP, vol. 6, 2016, pages 20948, XP055623863, DOI: 10.1038/srep20948
BODI, K. ET AL.: "Comparison of commercially available target enrichment methods for next-generation sequencing", J BIOMOL TECH, vol. 24, 2013, pages 73 - 86
NEWMAN, A. M. ET AL.: "Integrated digital error suppression for improved detection of circulating tumor DNA", NAT BIOTECHNOL, 2016
SAMORODNITSKY, E. ET AL.: "Evaluation of Hybridization Capture Versus Amplicon-Based Methods for Whole-Exome Sequencing", HUM MUTAT, vol. 36, 2015, pages 903 - 914
KUSCU, C.ARSLAN, S.SINGH, R.THORPE, J.ADLI, M.: "Genome-wide analysis reveals characteristics of off-target sites bound by the Cas9 endonuclease", NAT BIOTECHNOL, vol. 32, 2014, pages 677 - 683, XP055382577, DOI: 10.1038/nbt.2916
WU, X. ET AL.: "Genome-wide binding of the CRISPR endonuclease Cas9 in mammalian cells.", NAT BIOTECHNOL, vol. 32, 2014, pages 670 - 676, XP055241568, DOI: 10.1038/nbt.2889
LIU, X. ET AL.: "In Situ Capture of Chromatin Interactions by Biotinylated dCas9", CELL, vol. 170, 2017, pages 1028 - 1043
FUJITA, T.YUNO, M.FUJII, H.: "Efficient sequence-specific isolation of DNA fragments and chromatin by in vitro enChIP technology using recombinant CRISPR ribonucleoproteins", GENES CELLS, vol. 21, 2016, pages 370 - 377, XP055621734, DOI: 10.1111/gtc.12341
SWARTS, D. C. ET AL.: "The evolutionary journey of Argonaute proteins", NAT STRUCT MOL BIOL, vol. 21, 2014, pages 743 - 753, XP055287457, DOI: 10.1038/nsmb.2879
SONG, J. J.SMITH, S. K.HANNON, G. J.JOSHUA-TOR, L.: "Crystal structure of Argonaute and its implications for RISC slicer activity", SCIENCE, vol. 305, 2004, pages 1434 - 1437, XP002470480, DOI: 10.1126/science.1102514
SWARTS, D. C. ET AL.: "Argonaute of the archaeon Pyrococcus furiosus is a DNA-guided nuclease that targets cognate DNA", NUCLEIC ACIDS RES, vol. 43, 2015, pages 5120 - 5129, XP055287460, DOI: 10.1093/nar/gkv415
RAINES, R. T.MCCORMICK, M.VAN OOSBREE, T. R.MIERENDORF, R. C.: "The S.Tag fusion system for protein purification.", METHODS ENZYMOL, vol. 326, 2000, pages 362 - 376, XP009135533
Attorney, Agent or Firm:
KING & WOOD MALLESONS (CN)
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