Patent application EP19171002A (EP), filed 2019-04-25
Publication number: EP3730610A1
Title: MODIFIED CAS9 SYSTEM AND ITS USE FOR IMPROVED GENE EDITING
Abstract: The present invention relates to modified Cas9 nuclease comprising a substantial part of a Cas9 nuclease and covalently linked thereto at least one of a protein domain of a HDR factor selected from the group consisting of RAD51, RAD52, RAD54, MRE11, PALB2, FANCD2 and EXO1.
IPC codes: C12N  15/    62            A I
C12N   9/    22            A I
Claims: number: 1
text: Modified Cas9 nuclease comprising a substantial part of a Cas9 nuclease and fused thereto at least one of a protein domain of a HDR factor selected from the group consisting of RAD51, RAD52, RAD54, MRE11, PALB2, FANCD2 and EXO1.
number: 2
text: Modified Cas9 nuclease according to claim 1 characterized in that the Cas9 nuclease fragment is derived from Streptococcus pyogenes.
number: 3
text: Modified Cas9 nuclease according to claim 1 or 2 characterized in that the HDR factor is selected from RAD51, RAD52 and/or MRE11.
number: 4
text: Modified Cas9 nuclease according to claim 3 characterized in that it comprises at least two HDR factors selected from RAD51, RAD52 and MRE11.
number: 5
text: Modified Cas9 nuclease according to any of claims 1 to 4 characterized in that it comprises a hemagglutination tag and/or a promoter derived from CMV.
number: 6
text: Modified Cas9 nuclease according to claim 3 characterized in that it further comprises CtlP.
number: 7
text: Nucleic acid coding for a modified Cas9 nuclease according to any of claims 1 to 6.
number: 8
text: Nucleic acid according to claim 7 characterized in that it is a DNA sequence.
number: 9
text: Vector for the transfection of target cells characterized in that it comprises a nucleic acid according to claims 7 and 8.
number: 10
text: Vector according to claim 9 characterized in that it is a plasmid.
number: 11
text: Vector according to claim 9 characterized in that it is a vector comprising viral elements.
number: 12
text: Method for editing nucleic acid sequences in a homology directed repair wherein a modified Cas9 nuclease according to any of claims 1 to 6 and/or a nucleic acid according to claim 7 or 8 and/or a vector according to any of claims 10 and 11 is transferred into target cell.
Office action: search_report, dated 2020-10-28
Rejection reasons: novelty_epc_54
Prior art cited: ref_id: - CHARPENTIER ET AL., NATURE COMMUNICATIONS, vol. 9, 2018, pages 1133
ref_type: npl
severity: background
ref_id: WO2017142923A1
ref_type: patent
severity: background
ref_id: WO2016054326A1
ref_type: patent
severity: background
ref_id: WO2017180694A1
ref_type: patent
severity: background
ref_id: - SIMIN SHAO ET AL: "Enhancing CRISPR/Cas9-mediated homology-directed repair in mammalian cells by expressing Saccharomyces cerevisiae Rad52", INTERNATIONAL JOURNAL OF BIOCHEMISTRY AND CELL BIOLOGY, vol. 92, 18 September 2017 (2017-09-18), GB, pages 43 - 52, XP055627157, ISSN: 1357-2725, DOI: 10.1016/j.biocel.2017.09.012
ref_type: npl
severity: background
ref_id: - LING WANG ET AL: "Enhancing Targeted Genomic DNA Editing in Chicken Cells Using the CRISPR/Cas9 System", PLOS ONE, vol. 12, no. 1, 9 January 2017 (2017-01-09), pages e0169768, XP055385966, DOI: 10.1371/journal.pone.0169768
ref_type: npl
severity: background
ref_id: WO2016054326
ref_type: patent
severity: novelty_destroying
ref_id: WO2017142923
ref_type: patent
severity: novelty_destroying
ref_id: WO2017180694
ref_type: patent
severity: novelty_destroying
ref_id: - SIMIN SHAO ET AL, "Enhancing CRISPR/Cas9-mediated homology-directed repair in mammalian cells by expressing Saccharomyces cerevisiae Rad52", INTERNATIONAL JOURNAL OF BIOCHEMISTRY AND CELL BIOLOGY, GB, (20170918), vol. 92, doi:10.1016/j.biocel.2017.09.012, ISSN 1357-2725, pages 43 - 52, XP055627157 [X] 1-3,7-10,12 * the whole document *
ref_type: npl
severity: novelty_destroying
ref_id: - LING WANG ET AL, "Enhancing Targeted Genomic DNA Editing in Chicken Cells Using the CRISPR/Cas9 System", PLOS ONE, (20170109), vol. 12, no. 1, doi:10.1371/journal.pone.0169768, page e0169768, XP055385966 [X] 1-3,7-10,12 * page  9, paragraph  1  - paragraph  2; figures 4,5 * * page  2, paragraph  4  - paragraph  5 * * page  3, paragraph  4 *
ref_type: npl
severity: novelty_destroying
Final disposition: pending (2020-10-28)