Browsing by Author "Chao, Jiun-Shuan"

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Development of Single-Vector Tet-on Inducible Systems with High Sensitivity to Doxycycline
(2012-07-01)
Chao, Jiun-Shuan
;
Chao, Chun-Chieh
;
Chang, Chu-Li
;
Chiu, Yi-Rong
;
Yuan, Chiun-Jye
Single-vector Tet-on systems were developed to enable the tight regulation of transgenes in mammalian cells with a low dosage of doxycycline. Both the regulatory and the responsive units were integrated in a single vector and separated by a short DNA segment (214 bp). In the developed single-vector Tet-on systems, a high level of expression of the transgene can be induced by doxycycline at a concentration of as low as 1 ng/ml, which is 500-1,000 times lower than that usually utilized in other Tet-on systems. The single-vector Tet-on system developed here exhibited 3.5-10.8 times greater inducibility of the transgene in response to doxycycline than did a dual-vector system from a commercial source. Further studies indicate that the basal activity of Tet-on systems depends greatly on the strength of the promoter that controls the transactivator. The basal activity of Tet-on systems was high when the transactivator that was directed by the human cytomegalovirus promoter, and it was almost undetectable when the transactivator was placed under the control of a moderate strength mouse mammary tumor virus promoter. Moreover, the introduction of selectable markers allows the developed single-vector Tet-on systems to facilitate the generation of conditional transgenic cells and animals with high inducibility, low basal activity and detrimental effects of the long-term administration of doxycycline.
Enzyme-encapsulated silica nanoparticle for cancer chemotherapy
(2012-04-01)
Chiu, Yi-Rong
;
Ho, Wei-Jen
;
Chao, Jiun-Shuan
;
Yuan, Chiun-Jye
;
生物科技學系
A novel horseradish peroxidase-encapsulated silica nanoparticle (SNP) was generated in this study under relatively mild conditions. The generated enzyme-encapsulated SNP were relatively uniform in size (average 70 +/- 14.3 nm), monodispersed, and spherical, as characterized by transmission electron microscopy and scanning electron microscopy. The horseradish peroxidase encapsulated in silica nanoparticle exhibits biological properties, such as a pH-dependent activity profile and k(m) value, similar to that of free enzymes. Furthermore, enzyme-encapsulated SNP exhibited good operational stability for the repetitive usage with a relative standard deviation of 5.1 % ( n = 10) and a high stability for long term storage (>60 days) at 4 degrees C. The feasibility of using enzyme-encapsulated SNP in prodrug cancer therapy was also demonstrated by its capability to convert the prodrug indole-3-acetic acid into cytotoxic peroxyl radicals and trigger the death of tumor cells. These results indicate that the developed enzyme-encapsulated SNP has potential in the applications of prodrug cancer therapy.