Transient transfection using 222 nm far UV-C irradiation
Abstract Ultraviolet (UV) light is never used for gene transfer because it damages DNA and harms cellular and plasmid DNA. A light source selectively radiating Far UV-C (F-UV) light causes less DNA damage. We investigated potential introduction of plasmids into cells by irradiating them with F-UV li...
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Nature Portfolio
2025-05-01
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| Online Access: | https://doi.org/10.1038/s41598-025-00477-6 |
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| author | Mane Nishimura Yuki Shimizu Tomohiro Fujii Yu Okada Takeshi Yamamoto Yoshimasa Ogawa Toru Koi Yutaka Suehiro Takahiro Yamasaki Jun Nishikawa |
| author_facet | Mane Nishimura Yuki Shimizu Tomohiro Fujii Yu Okada Takeshi Yamamoto Yoshimasa Ogawa Toru Koi Yutaka Suehiro Takahiro Yamasaki Jun Nishikawa |
| author_sort | Mane Nishimura |
| collection | DOAJ |
| description | Abstract Ultraviolet (UV) light is never used for gene transfer because it damages DNA and harms cellular and plasmid DNA. A light source selectively radiating Far UV-C (F-UV) light causes less DNA damage. We investigated potential introduction of plasmids into cells by irradiating them with F-UV light. COS-7 and CHO-K1 cells were irradiated with 222 nm F-UV light. Then, DNA solution containing green fluorescent protein (EGFP) plasmid was added to the culture and incubated (37 °C, 5% CO2, 24 h) for fluorescence microscopy. Cytotoxicity of cells irradiated with the same energy as for gene transfer was evaluated. Characteristics of EGFP-positive cells were compared with non-transfected cells by propidium iodide (PI) and Hoechst staining. Cells with distinct green fluorescence were observed after F-UV light irradiation and addition of 200 ng of EGFP plasmid. COS-7 cells showed the highest number of EGFP-positive cells at 0.5 mJ/cm2 irradiation, whereas that for CHO-K1 cells was at 1 mJ/cm2 irradiation. Cytotoxicity was low in COS-7 at ≤ 1 mJ/cm2 irradiation and CHO-K1 at ≤ 0.5 mJ/cm2. Transfected cells did not incorporate PI, and their nuclei did not differ from those of non-transfected cells. We successfully transfected two cell lines with EGFP plasmids by F-UV irradiation. |
| format | Article |
| id | doaj-art-c4f9d06f8301439db97fe9b16e13d4a7 |
| institution | DOAJ |
| issn | 2045-2322 |
| language | English |
| publishDate | 2025-05-01 |
| publisher | Nature Portfolio |
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| series | Scientific Reports |
| spelling | doaj-art-c4f9d06f8301439db97fe9b16e13d4a72025-08-20T03:10:17ZengNature PortfolioScientific Reports2045-23222025-05-011511610.1038/s41598-025-00477-6Transient transfection using 222 nm far UV-C irradiationMane Nishimura0Yuki Shimizu1Tomohiro Fujii2Yu Okada3Takeshi Yamamoto4Yoshimasa Ogawa5Toru Koi6Yutaka Suehiro7Takahiro Yamasaki8Jun Nishikawa9Faculty of Laboratory Science, Yamaguchi University Graduate School of MedicineFaculty of Laboratory Science, Yamaguchi University Graduate School of MedicineFaculty of Laboratory Science, Yamaguchi University Graduate School of MedicineFaculty of Laboratory Science, Yamaguchi University Graduate School of MedicineFaculty of Laboratory Science, Yamaguchi University Graduate School of MedicineUshio Inc.Ushio Inc.Department of Oncology and Laboratory Medicine, Yamaguchi University Graduate School of MedicineDepartment of Oncology and Laboratory Medicine, Yamaguchi University Graduate School of MedicineFaculty of Laboratory Science, Yamaguchi University Graduate School of MedicineAbstract Ultraviolet (UV) light is never used for gene transfer because it damages DNA and harms cellular and plasmid DNA. A light source selectively radiating Far UV-C (F-UV) light causes less DNA damage. We investigated potential introduction of plasmids into cells by irradiating them with F-UV light. COS-7 and CHO-K1 cells were irradiated with 222 nm F-UV light. Then, DNA solution containing green fluorescent protein (EGFP) plasmid was added to the culture and incubated (37 °C, 5% CO2, 24 h) for fluorescence microscopy. Cytotoxicity of cells irradiated with the same energy as for gene transfer was evaluated. Characteristics of EGFP-positive cells were compared with non-transfected cells by propidium iodide (PI) and Hoechst staining. Cells with distinct green fluorescence were observed after F-UV light irradiation and addition of 200 ng of EGFP plasmid. COS-7 cells showed the highest number of EGFP-positive cells at 0.5 mJ/cm2 irradiation, whereas that for CHO-K1 cells was at 1 mJ/cm2 irradiation. Cytotoxicity was low in COS-7 at ≤ 1 mJ/cm2 irradiation and CHO-K1 at ≤ 0.5 mJ/cm2. Transfected cells did not incorporate PI, and their nuclei did not differ from those of non-transfected cells. We successfully transfected two cell lines with EGFP plasmids by F-UV irradiation.https://doi.org/10.1038/s41598-025-00477-6Far UV-C222 nmTransfectionDeep-ultravioletDNA damage |
| spellingShingle | Mane Nishimura Yuki Shimizu Tomohiro Fujii Yu Okada Takeshi Yamamoto Yoshimasa Ogawa Toru Koi Yutaka Suehiro Takahiro Yamasaki Jun Nishikawa Transient transfection using 222 nm far UV-C irradiation Scientific Reports Far UV-C 222 nm Transfection Deep-ultraviolet DNA damage |
| title | Transient transfection using 222 nm far UV-C irradiation |
| title_full | Transient transfection using 222 nm far UV-C irradiation |
| title_fullStr | Transient transfection using 222 nm far UV-C irradiation |
| title_full_unstemmed | Transient transfection using 222 nm far UV-C irradiation |
| title_short | Transient transfection using 222 nm far UV-C irradiation |
| title_sort | transient transfection using 222 nm far uv c irradiation |
| topic | Far UV-C 222 nm Transfection Deep-ultraviolet DNA damage |
| url | https://doi.org/10.1038/s41598-025-00477-6 |
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