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Construction and functional validation of cytoplasmic domain-deleted human CD2 (hCD2ΔCyto) as a selection marker for retroviral transduction
http://hdl.handle.net/10129/0002001987
http://hdl.handle.net/10129/000200198781de956b-6ab6-4596-877b-b3b885682d35
| 名前 / ファイル | ライセンス | アクション |
|---|---|---|
Hirosaki Med J 76-1-4_39.pdf (2.9 MB)
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| アイテムタイプ | リポジトリ登録用アイテムタイプ（フル）(1) | |||||||||||||
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| 公開日 | 2026-03-25 | |||||||||||||
| タイトル | ||||||||||||||
| タイトル | Construction and functional validation of cytoplasmic domain-deleted human CD2 (hCD2ΔCyto) as a selection marker for retroviral transduction | |||||||||||||
| 言語 | en | |||||||||||||
| 作成者 |
Odagiri, Haruka
× Odagiri, Haruka
× Fuji, Hodaka
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| アクセス権 | ||||||||||||||
| アクセス権 | open access | |||||||||||||
| アクセス権URI | http://purl.org/coar/access_right/c_abf2 | |||||||||||||
| 内容記述 | ||||||||||||||
| 内容記述タイプ | Abstract | |||||||||||||
| 内容記述 | Human CD2 （hCD2） is used widely as a surface selection marker during gene transfer experiments because of its strong expression and compatibility with flow cytometry analysis and magnetic cell sorting; however, its cytoplasmic domain contains signaling motifs that may inadvertently affect cellular function, particularly that of immune cells. To resolve this potential problem, we developed a truncated version of hCD2 lacking the intracellular signaling domain （hCD2ΔCyto） and evaluated its performance as a selection marker. The hCD2ΔCyto construct was generated by PCR and cloned into a retroviral vector using Gibson assembly. Sequence integrity was confirmed by plasmid sequencing, and the construct was transduced into 293T cells. Flow cytometry analysis revealed detectable surface expression of hCD2ΔCyto, and magnetic sorting following transduction allowed efficient enrichment of hCD2-positive cells. Western blot analysis confirmed expression of proteins, with broad bands likely attributable to glycosylation. Furthermore, co- expression of EGFP using a bicistronic IRES-based vector demonstrated compatibility with polycistronic systems. These findings suggest that hCD2ΔCyto can serve as a selection marker for cells without detectable signaling activity. While additional validation in immune cell types is needed, this construct provides a promising alternative to full-length hCD2 for applications requiring minimal perturbation of cellular function. | |||||||||||||
| 言語 | en | |||||||||||||
| 出版者 | ||||||||||||||
| 出版者 | 弘前大学大学院医学研究科 | |||||||||||||
| 言語 | ja | |||||||||||||
| 出版者 | ||||||||||||||
| 出版者 | Hirosaki University Graduate School of Medicine | |||||||||||||
| 言語 | en | |||||||||||||
| 言語 | ||||||||||||||
| 言語 | eng | |||||||||||||
| 資源タイプ | ||||||||||||||
| 資源タイプ識別子 | http://purl.org/coar/resource_type/c_6501 | |||||||||||||
| 資源タイプ | journal article | |||||||||||||
| 出版タイプ | ||||||||||||||
| 出版タイプ | VoR | |||||||||||||
| 出版タイプResource | http://purl.org/coar/version/c_970fb48d4fbd8a85 | |||||||||||||
| 収録物識別子 | ||||||||||||||
| 収録物識別子タイプ | NCID | |||||||||||||
| 収録物識別子 | AN00211444 | |||||||||||||
| 収録物識別子 | ||||||||||||||
| 収録物識別子タイプ | PISSN | |||||||||||||
| 収録物識別子 | 0439-1721 | |||||||||||||
| 収録物識別子 | ||||||||||||||
| 収録物識別子タイプ | EISSN | |||||||||||||
| 収録物識別子 | 2434-4656 | |||||||||||||
| 書誌情報 |
ja : 弘前医学 en : Hirosaki Medical Journal 巻 76, 号 1-4, p. 39-47, 発行日 2026-03-10 |
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