Chiba Mayumi, Kamiya Mako, Tsuda-Sakurai Kayoko, Fujisawa Yuya, Kosakamoto Hina, Kojima Ryosuke, Miura Masayuki, Urano Yasuteru
Graduate School of Medicine and Graduate School of Pharmaceutical Sciences, The University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-0033, Japan.
PRESTO, Japan Science and Technology Agency, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan.
ACS Cent Sci. 2019 Oct 23;5(10):1676-1681. doi: 10.1021/acscentsci.9b00678. Epub 2019 Aug 26.
To achieve highly selective ablation of -positive cells in a biological milieu , we developed an activatable photosensitizer, SPiDER-killer-βGal, targeted to β-galactosidase encoded by the reporter gene. Hydrolysis of SPiDER-killer-βGal by β-galactosidase simultaneously activates both its photosensitizing ability and its reactivity to nucleophiles, so that the phototoxic products generated by light irradiation are trapped inside the -positive cells. The combination of SPiDER-killer-βGal and light irradiation specifically killed -positive cells in coculture with cells without expression. Furthermore, β-galactosidase-expressing cells in the posterior region of cultured wing discs and in pupal notum of live pupae were selectively killed with single-cell resolution. This photosensitizer should be useful for specific ablation of targeted cells in living organisms, for example, to investigate cellular functions in complex networks.
为了在生物环境中实现对β-半乳糖苷酶阳性细胞的高度选择性消融，我们开发了一种可激活的光敏剂SPiDER-killer-βGal，它靶向报告基因编码的β-半乳糖苷酶。β-半乳糖苷酶对SPiDER-killer-βGal的水解同时激活了其光敏能力和对亲核试剂的反应性，从而使光照射产生的光毒性产物被困在β-半乳糖苷酶阳性细胞内。SPiDER-killer-βGal与光照射相结合，特异性地杀死了与无β-半乳糖苷酶表达的细胞共培养的β-半乳糖苷酶阳性细胞。此外，培养的果蝇翅芽后部区域和活体果蝇蛹期背板中表达β-半乳糖苷酶的细胞以单细胞分辨率被选择性杀死。这种光敏剂应该有助于在活生物体中特异性消融靶向细胞，例如，用于研究复杂网络中的细胞功能。
