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首页> 外文期刊>Proceedings of the National Academy of Sciences of the United States of America >A mutant herpes simplex virus type 1 thymidine kinase reporter gene shows improved sensitivity for imaging reporter gene expression with positron emission tomography
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A mutant herpes simplex virus type 1 thymidine kinase reporter gene shows improved sensitivity for imaging reporter gene expression with positron emission tomography

机译：突变的单纯疱疹病毒1型胸苷激酶报告基因突变体显示正电子发射断层显像成像基因表达的敏感性提高

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摘要

We are developing assays for noninvasive. quantitative imaging of reporter genes with positron emission tomography (PET), for application both in animal models and in human gene therapy. We report here a method to improve the detection of lower levels of PET reporter gene expression by utilizing a mutant herpes simplex virus type 1 thymidine kinase (HSV1-sr39tk) as a PET reporter gene. The HSV1-sr39tk mutant was identified from a ;library of site- directed mutants. Accumulation (net uptake) of the radioactively labeled substrates l8-3H]penciclovir ([8-3HlPCV). and 8-l18F]fluoro- penciclovir (FPCV) in C6 rat glioma cells expressing HSV1-sr39tk is increased by a factor of rs2.0 when compared with C6 cells expressing wild-type HSV1-tk. The increased imaging sensitivity of HSV1-sr39tk when FPCV is used is also demonstrated in vivo both with tumor cells stably transfected with either HSV1-tk or HSV1- sr39tk, and after hepatic delivery of HSV1-tk or HSV1-sr39tk by using adenoviral vectors. The use of HSV1-sr39tk as a PET reporter gene and FPCV as a PET reporter probe results in significantly enhanced sensitivity for imaging reporter gene expression in vivo.
机译：我们正在开发无创检测方法。利用正电子发射断层扫描（PET）对报告基因进行定量成像，可用于动物模型和人类基因治疗。我们在这里报告了一种方法，可通过利用突变型单纯疱疹病毒1型胸苷激酶（HSV1-sr39tk）作为PET报告基因来提高检测PET报告基因较低水平的方法。 HSV1-sr39tk突变体是从定点突变体文库中鉴定的。放射性标记底物18-3H]更昔洛韦（[8-3H1PCV）的积累（净吸收）。与表达野生型HSV1-tk的C6细胞相比，表达HSV1-sr39tk的C6大鼠神经胶质瘤细胞中的8和18-18F]氟-喷昔洛韦（FPCV）增加rs2.0倍。使用FPCV时，HSV1-sr39tk的影像学敏感性也得到了提高，无论是用HSV1-tk或HSV1-sr39tk稳定转染的肿瘤细胞，还是通过腺病毒载体肝转移HSV1-tk或HSV1-sr39tk的体内。 HSV1-sr39tk作为PET报告基因，FPCV作为PET报告探针的使用，可以显着增强体内成像报告基因表达的敏感性。

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