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<p>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&#xa0;nm F-UV light. Then, DNA solution containing green fluorescent protein (EGFP) plasmid was added to the culture and incubated (37&#xa0;°C, 5% CO<sub>2</sub>, 24&#xa0;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&#xa0;mJ/cm<sup>2</sup> irradiation, whereas that for CHO-K1 cells was at 1&#xa0;mJ/cm<sup>2</sup> irradiation. Cytotoxicity was low in COS-7 at ≤ 1&#xa0;mJ/cm<sup>2</sup> irradiation and CHO-K1 at ≤ 0.5&#xa0;mJ/cm<sup>2</sup>. 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.</p>

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Transient transfection using 222 nm far UV-C irradiation

  • 期刊论文

  • 出版模式：

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  • 发表日期：

    2025年5月14日

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  • Mane Nishimura,
  • Yuki Shimizu,
  • Tomohiro Fujii,
  • Yu Okada,
  • Takeshi Yamamoto,
  • Yoshimasa Ogawa,
  • Toru Koi,
  • Yutaka Suehiro,
  • Takahiro Yamasaki,
  • Jun Nishikawa

摘要

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.