GENE-EXPRESSION FOLLOWING DIRECT-INJECTION OF DNA INTO LIVER
GENE-EXPRESSION FOLLOWING DIRECT-INJECTION OF DNA INTO LIVER
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DOI：
10.1089/hum.1994.5.12-1477
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发表时间：
1994-12-01
影响因子：
4.2
通讯作者：
POWELL, JS
中科院分区：
文献类型：
--
作者：
HICKMAN, MA;MALONE, RW;POWELL, JS
肝脏是基因治疗的靶组织，目前的肝脏基因递送方法包括逆转录病毒载体和腺病毒载体、脂质体/DNA和肽/DNA复合物。本研究描述了直接将DNA注射到肝脏中导致显著基因表达的技术。在注射编码荧光素酶基因（pCMVL）、β-半乳糖苷酶（pCMVL）使用编码Lac Z的质粒DNA原位评估（β-Gal）活性（pCMV β），并在注射编码该蛋白质的质粒DNA后测量分泌的人α-1-抗胰蛋白酶的血清浓度pRC/CMV-sHAT转染肝细胞后，荧光素酶在24-48 h表达量最高，β-Gal染色显示大部分肝细胞位于注射部位附近。在注射pRC/CMV-sHAT的动物的血清中检测到显著浓度的人α-1-抗胰蛋白酶。这些发现证明了将质粒DNA直接注射到肝脏中可以导致显著的基因表达的一般原理。
The liver is an attractive target tissue for gene therapy, Current approaches for hepatic gene delivery include retroviral and adenoviral vectors, liposome/DNA, and peptide/DNA complexes, This study describes a technique for direct injection of DNA into liver that led to significant gene expression, Gene expression was characterized in both rats and cats following injection of plasmid DNA encoding several different proteins, Luciferase activity was measured after injection of plasmid DNA encoding the luciferase gene (pCMVL), beta-galactosidase (beta-Gal) activity was evaluated in situ using plasmid DNA encoding Lac Z (pCMV beta), and serum concentration of secreted human alpha-1-antitrypsin was measured following injection of plasmid DNA encoding this protein (pRC/CMV-sHAT), Several variables, including injection technique, DNA dose, and DNA diluent, were investigated, Direct injection of pCMVL resulted in maximal luciferase expression at 24-48 hr. beta-Gal staining demonstrated that the majority of transfected hepatocytes were located near the injection site. Significant concentrations of human alpha-1-antitrypsin were detected in the serum of animals injected with pRC/CMV-sHAT, These findings demonstrate the general principle that direct injection of plasmid DNA into liver can lead to significant gene expression.