Invention Publication
- Patent Title: LINEAR DNA WITH ENHANCED RESISTANCE AGAINST EXONUCLEASES AND METHODS FOR THE PRODUCTION THEREOF
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Application No.: US18429780Application Date: 2024-02-01
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Publication No.: US20240360488A1Publication Date: 2024-10-31
- Inventor: Heikki Lanckriet , Amy Walker , Ángel Picher
- Applicant: 4BASEBIO, S.L.U. , 4BASEBIO UK LTD
- Applicant Address: ES Madrid
- Assignee: 4BASEBIO, S.L.U.,4BASEBIO UK LTD
- Current Assignee: 4BASEBIO, S.L.U.,4BASEBIO UK LTD
- Current Assignee Address: ES Madrid
- Priority: EP 382083.6 2023.02.01
- Main IPC: C12P19/34
- IPC: C12P19/34 ; C12N9/22 ; C12N15/10

Abstract:
Methods for producing a linear deoxyribonucleic acid (DNA) product with enhanced resistance to nuclease digestion are provided. The methods comprise, (a) digesting a double-stranded DNA molecule with an endonuclease that cleaves an endonuclease target sequence to generate a digested double-stranded DNA molecule, wherein the digested double-stranded DNA molecule comprises a linear double-stranded region, and a truncated protelomerase sequence at a first end, wherein the truncated protelomerase target sequence is non-functional; (b) appending a first adaptor molecule to the first end of the digested double-stranded DNA molecule and appending a second adaptor molecule to the second end of the digested double-stranded DNA molecule to generate a precursor double-stranded DNA molecule, wherein the first adaptor molecule comprises a truncated protelomerase target sequence that forms a first functional protelomerase target sequence with the truncated protelomerase sequence at the first end of the digested double-stranded DNA molecule; and (c) incubating the precursor double-stranded DNA molecule with a protelomerase to generate the linear DNA product, wherein the protelomerase closes the first end of the precursor double-stranded DNA molecule at the first functional protelomerase target sequence.
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