Ministring DNA (msDNA): a novel linear covalently-closed DNA with enhanced stability for gene and cell therapy applications

Abstract The quality and fidelity of DNA vectors used in genetic medicine and gene therapy either as starting material for manufacturing or as therapeutic ingredients play a critical role in determining ultimate clinical success. Ministring DNA (msDNA), is a novel minivector that is a linear covalen...

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Main Authors: Wai Kuen Hung, Siddarth Chandrasekaran, Olga Zaslaver, Ming Zhu, Jamie Lam, Steven Hersch, Peyman Mokarami, Roderick A. Slavcev, Nafiseh Nafissi
Format: Article
Language:English
Published: Nature Portfolio 2025-05-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-98730-5
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author Wai Kuen Hung
Siddarth Chandrasekaran
Olga Zaslaver
Ming Zhu
Jamie Lam
Steven Hersch
Peyman Mokarami
Roderick A. Slavcev
Nafiseh Nafissi
author_facet Wai Kuen Hung
Siddarth Chandrasekaran
Olga Zaslaver
Ming Zhu
Jamie Lam
Steven Hersch
Peyman Mokarami
Roderick A. Slavcev
Nafiseh Nafissi
author_sort Wai Kuen Hung
collection DOAJ
description Abstract The quality and fidelity of DNA vectors used in genetic medicine and gene therapy either as starting material for manufacturing or as therapeutic ingredients play a critical role in determining ultimate clinical success. Ministring DNA (msDNA), is a novel minivector that is a linear covalently-closed (LCC) double-stranded DNA molecule devoid of immunogenic bacterial sequences (e.g., origin of replication, antibiotic resistant cassette). Similar to traditional plasmids, msDNA is manufactured in vivo in E. coli and therefore benefits from the scalability of E. coli -based systems and the ~ 1000-fold enhanced fidelity conferred by the mismatch repair (MMR) mechanism. In this paper, we address the improved stability of msDNA. We show that due to the torsion-free structure, msDNA is more stable to chemical and mechanical stress than conventional plasmid DNA. Moreover, we demonstrate that lyophilization can further improve the long-term stability of msDNA, reducing the need for cold chain storage. Therefore, we propose that msDNA can be a new paradigm for genetic medicine by offering genetic material with lower immunogenicity, reduced risk of insertional mutagenesis, and higher fidelity and stability.
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spelling doaj-art-7947fbedfaaa40e1b1b66e25f7e028d42025-08-20T01:47:29ZengNature PortfolioScientific Reports2045-23222025-05-0115111410.1038/s41598-025-98730-5Ministring DNA (msDNA): a novel linear covalently-closed DNA with enhanced stability for gene and cell therapy applicationsWai Kuen Hung0Siddarth Chandrasekaran1Olga Zaslaver2Ming Zhu3Jamie Lam4Steven Hersch5Peyman Mokarami6Roderick A. Slavcev7Nafiseh Nafissi8Mediphage BioceuticalsMediphage BioceuticalsMediphage BioceuticalsMediphage BioceuticalsMediphage BioceuticalsMediphage BioceuticalsMediphage BioceuticalsMediphage BioceuticalsMediphage BioceuticalsAbstract The quality and fidelity of DNA vectors used in genetic medicine and gene therapy either as starting material for manufacturing or as therapeutic ingredients play a critical role in determining ultimate clinical success. Ministring DNA (msDNA), is a novel minivector that is a linear covalently-closed (LCC) double-stranded DNA molecule devoid of immunogenic bacterial sequences (e.g., origin of replication, antibiotic resistant cassette). Similar to traditional plasmids, msDNA is manufactured in vivo in E. coli and therefore benefits from the scalability of E. coli -based systems and the ~ 1000-fold enhanced fidelity conferred by the mismatch repair (MMR) mechanism. In this paper, we address the improved stability of msDNA. We show that due to the torsion-free structure, msDNA is more stable to chemical and mechanical stress than conventional plasmid DNA. Moreover, we demonstrate that lyophilization can further improve the long-term stability of msDNA, reducing the need for cold chain storage. Therefore, we propose that msDNA can be a new paradigm for genetic medicine by offering genetic material with lower immunogenicity, reduced risk of insertional mutagenesis, and higher fidelity and stability.https://doi.org/10.1038/s41598-025-98730-5KeywordsMinistring DNALinear covalently closed DNAGene therapyChemical and mechanical stabilityLyophilization
spellingShingle Wai Kuen Hung
Siddarth Chandrasekaran
Olga Zaslaver
Ming Zhu
Jamie Lam
Steven Hersch
Peyman Mokarami
Roderick A. Slavcev
Nafiseh Nafissi
Ministring DNA (msDNA): a novel linear covalently-closed DNA with enhanced stability for gene and cell therapy applications
Scientific Reports
Keywords
Ministring DNA
Linear covalently closed DNA
Gene therapy
Chemical and mechanical stability
Lyophilization
title Ministring DNA (msDNA): a novel linear covalently-closed DNA with enhanced stability for gene and cell therapy applications
title_full Ministring DNA (msDNA): a novel linear covalently-closed DNA with enhanced stability for gene and cell therapy applications
title_fullStr Ministring DNA (msDNA): a novel linear covalently-closed DNA with enhanced stability for gene and cell therapy applications
title_full_unstemmed Ministring DNA (msDNA): a novel linear covalently-closed DNA with enhanced stability for gene and cell therapy applications
title_short Ministring DNA (msDNA): a novel linear covalently-closed DNA with enhanced stability for gene and cell therapy applications
title_sort ministring dna msdna a novel linear covalently closed dna with enhanced stability for gene and cell therapy applications
topic Keywords
Ministring DNA
Linear covalently closed DNA
Gene therapy
Chemical and mechanical stability
Lyophilization
url https://doi.org/10.1038/s41598-025-98730-5
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