Functional Coexpression of HSV-1 Thymidine Kinase and Green Fluorescent Protein: Implications for Noninvasive Imaging of Transgene Expression

摘要: Abstract Current gene therapy technology is limited by the paucity of methodology for determining location and magnitude therapeutic transgene expression in vivo . We describe validate a paradigm monitoring noninvasive imaging herpes simplex virus type 1 thymidine kinase (HSV-1- tk ) marker expression. To test proportional coexpression genes, model fusion comprising green fluorescent protein (gfp) HSV-1- genes was generated ( tkgfp gene) assessed functional product, TKGFP protein, rat 9L gliosarcoma, RG2 glioma, W256 carcinoma cells. Analysis demonstrated that it can serve as rendering transduced cells sensitive to ganciclovir or screening useful identifying fluorescence microscopy fluorescence-activated cell sorting (FACS). TK GFP activities were similar corresponding wild-type proteins accumulation -specific radiolabeled substrate, 2′-fluoro-2′-deoxy-1β-D-arabino-furanosyl-5-iodo-uracil (FIAU), stability clones correlated with gfp-fluorescence intensity over wide range levels. The itself may be developing novel cancer approaches. Valuable information about efficiency transfer could obtained non-invasive FIAU clinical devices (gamma camera, positron-emission tomography [PET], single photon emission computed [SPECT]), and/or direct visualization gfp situ endoscopy.