원문정보
Liposome-Mediated Electric Gene Delivery into Fetal and Adult Gonads
초록
영어
Gene delivery is one of the keen interests in animal industry as well as research on gene functions. Some of the in vivo gene delivery techniques have been successively used in various tissues for the gene therapy and transgenesis. Despite intensive efforts, it still remains to overcome problems of limited local and regional administration and low transgene expression. To improve the efficiency of gene delivery, a new procedure was tested. We injected exogenous DNA containing LacZ into the female or male gonads and then pulsed electric field. Electroporated gonads showed positive X-gal staining in many seminiferous tubules of the porcine fetal gonads. Exogenously introduced LacZ genes were also expressed in female porcine gonad. In addition, we demonstrated efficient gene delivery in gonad of adult mouse. Furthermore, we succeed to generate genetically modified germline cells showing GFP and positive X-gal signals. The results suggest that the newly developed gene delivery is an effective way of in vivo transfection in mammals. The developed gene delivery procedure should be useful in producing transgenic animals when combined with primary cell culture and nuclear transplantation.
목차
II. MATERIALS AND METHODS
1. Preparation of plasmid DNA and DNA/ liposome complexes
2. Animals and in vivo gene transfer
3. Culture and gene transfection
4. X-gal staining of transfected cells or tissues
III. RESULTS
1. lsolation and Characterization of Gonadal Sex from Porcine Fetuses
2. Expression of Transgene Introduced into the Gonads
3. In vitro Gene Expression in Porcine Primary Gonad Cells
IV. DISCUSSION
V. REFERENCES