Brain protection and recirculation therapy with non-viral vector gene

1）为了建立一种能特异性分泌β-内啡肽的细胞系，将人β-内啡肽编码序列与NL 1基因的一部分连接，构建了一种新的融合基因。转染该融合基因的Tet-on系统的HEK 293细胞在给予强力霉素（Dox）后以剂量依赖性方式分泌β-内啡肽。将该细胞系包封并移植到猴的CSF空间中，但对施用的Dox（i.m.）没有反应。另一方面，将Neuro 2A-Tet-on-POMC细胞包封并移植到猴的CSF空间中。它们在给予Dox后以剂量依赖性方式在猴的CSF中分泌ACTH。根据这些结果，可以通过Tet-on系统控制CSF空间中的包囊细胞的肽分泌。HEK 293细胞可能不适合本实验。2）将含有lacZ、GFP-ODN或DsRed的HVJ-E注射到大鼠和猴的枕大池中，并用4%多聚甲醛灌注。他们的大脑和脑膜被移除，并制作了50 μ m厚的切片。分别用lacZ、GFP-ODN和DsRed转染猴和大鼠的脑和脑膜。基因转染的功效在两种动物中均不稳定。其原因尚不清楚，也未做生理检查。

1)To create a cell line with controlled and specific secretion of b-endorphin, a new fusion gene was constructed by joining human b-endorphin coding sequence to part of NL1 gene. HEK293 cells carrying Tet-on system transfected with this fusion gene secreted b-endorphin in a dose-dependent manner upon doxycycline(Dox) administration. This cell line was encapsulated and transplanted in the monkey's CSF space, but failed to respond the administered Dox (i.m.). On the other hand, Neuro2A-Tet-on-POMC cells were encapsulated and transplanted in the monkey's CSF space. They secreted ACTH in the monkey's CSF in a dose-dependent manner upon Dox administration. From these results, peptide secretion from encapsulated cells in the CSF space can be controlled through Tet-on system. HEK293 cells may not fit for this experiment.2)HVJ-E containing the lacZ, GFP-ODN or DsRed was injected into the cisterna magna of rats and monkeys, and they were perfused with 4% paraformaldehyde. Their brains and meninges were removed and 50-μm-thick sections were made. The brains and meninges of both monkeys and rats were examined about the gene transfection of lacZ, GFP-ODN or DsRed. The efficacy of gene transfection was not stable in both animals. It's reason wasnot clear and the physiological examination has not been done.