High spontaneous integration rates of end-modified linear DNAs upon mammalian cell transfection
Abstract In gene therapy, potential integration of therapeutic transgene into host cell genomes is a serious risk that can lead to insertional mutagenesis and tumorigenesis. Viral vectors are often used as the gene delivery vehicle, but they are prone to undergoing integration events. More recently,...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Nature Portfolio
2023-04-01
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| Series: | Scientific Reports |
| Online Access: | https://doi.org/10.1038/s41598-023-33862-0 |
| _version_ | 1797836455162150912 |
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| author | Samuel Lim R. Rogers Yocum Pamela A. Silver Jeffrey C. Way |
| author_facet | Samuel Lim R. Rogers Yocum Pamela A. Silver Jeffrey C. Way |
| author_sort | Samuel Lim |
| collection | DOAJ |
| description | Abstract In gene therapy, potential integration of therapeutic transgene into host cell genomes is a serious risk that can lead to insertional mutagenesis and tumorigenesis. Viral vectors are often used as the gene delivery vehicle, but they are prone to undergoing integration events. More recently, non-viral delivery of linear DNAs having modified geometry such as closed-end linear duplex DNA (CELiD) have shown promise as an alternative, due to prolonged transgene expression and less cytotoxicity. However, whether modified-end linear DNAs can also provide a safe, non-integrating gene transfer remains unanswered. Herein, we compare the genomic integration frequency upon transfection of cells with expression vectors in the forms of circular plasmid, unmodified linear DNA, CELiDs with thioester loops, and Streptavidin-conjugated blocked-end linear DNA. All of the forms of linear DNA resulted in a high fraction of the cells being stably transfected—between 10 and 20% of the initially transfected cells. These results indicate that blocking the ends of linear DNA is insufficient to prevent integration. |
| first_indexed | 2024-04-09T15:10:14Z |
| format | Article |
| id | doaj.art-fabe5d356c4b46a5b12156382aca5cc0 |
| institution | Directory Open Access Journal |
| issn | 2045-2322 |
| language | English |
| last_indexed | 2024-04-09T15:10:14Z |
| publishDate | 2023-04-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Scientific Reports |
| spelling | doaj.art-fabe5d356c4b46a5b12156382aca5cc02023-04-30T11:16:38ZengNature PortfolioScientific Reports2045-23222023-04-011311810.1038/s41598-023-33862-0High spontaneous integration rates of end-modified linear DNAs upon mammalian cell transfectionSamuel Lim0R. Rogers Yocum1Pamela A. Silver2Jeffrey C. Way3Department of Systems Biology, Harvard Medical SchoolGeneral Biologics, IncDepartment of Systems Biology, Harvard Medical SchoolGeneral Biologics, IncAbstract In gene therapy, potential integration of therapeutic transgene into host cell genomes is a serious risk that can lead to insertional mutagenesis and tumorigenesis. Viral vectors are often used as the gene delivery vehicle, but they are prone to undergoing integration events. More recently, non-viral delivery of linear DNAs having modified geometry such as closed-end linear duplex DNA (CELiD) have shown promise as an alternative, due to prolonged transgene expression and less cytotoxicity. However, whether modified-end linear DNAs can also provide a safe, non-integrating gene transfer remains unanswered. Herein, we compare the genomic integration frequency upon transfection of cells with expression vectors in the forms of circular plasmid, unmodified linear DNA, CELiDs with thioester loops, and Streptavidin-conjugated blocked-end linear DNA. All of the forms of linear DNA resulted in a high fraction of the cells being stably transfected—between 10 and 20% of the initially transfected cells. These results indicate that blocking the ends of linear DNA is insufficient to prevent integration.https://doi.org/10.1038/s41598-023-33862-0 |
| spellingShingle | Samuel Lim R. Rogers Yocum Pamela A. Silver Jeffrey C. Way High spontaneous integration rates of end-modified linear DNAs upon mammalian cell transfection Scientific Reports |
| title | High spontaneous integration rates of end-modified linear DNAs upon mammalian cell transfection |
| title_full | High spontaneous integration rates of end-modified linear DNAs upon mammalian cell transfection |
| title_fullStr | High spontaneous integration rates of end-modified linear DNAs upon mammalian cell transfection |
| title_full_unstemmed | High spontaneous integration rates of end-modified linear DNAs upon mammalian cell transfection |
| title_short | High spontaneous integration rates of end-modified linear DNAs upon mammalian cell transfection |
| title_sort | high spontaneous integration rates of end modified linear dnas upon mammalian cell transfection |
| url | https://doi.org/10.1038/s41598-023-33862-0 |
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