中华耳科学杂志2011，Vol.9Issue(3)：258-263,6.
内耳相关基础研究专辑基于聚酰胺胺的非病毒载体携带Math1基因进行体外基因转导的有效性和安全性评价
Efficiency and safety of Math1 gene in vitro transduction with a PAMAM dentrimers-based non-viral vector
摘要
Abstract
Objective To develop a novel non-viral gene vector based on PAMAM dendrimers that is able to transduce the Math1 gene, and to evaluate its efficiency and safety. Methods PAMAM was modified by cyclodextrin with different weight ratios (1:2, 1:1,4:1 or 8:1) after activation at 60 degree for 6, 12, 24 or 48 hours. Polyplex was formed by mixing PAMAM solutions with plasmid solution containing pRK5-Mathl-EGFP of different N/P ratios (1/1, 2/ 1, 5/1 or 10/1). Transduction was performed using HEK-239T cells under non-serum conditions. The efficiency of transduction was evaluated using percentage of EGFP positive cells, while the safety was estimated using MIT. Results The maximum transduction efficiency (38.14 ± 2.85%) was achieved, with a 68.86 ± 3.44% viability, when vectors were heated for 24 hours, with a 1 : 1 PAMAM : CD ratio and 5/1 N/P ratio. Conclusion We have developed a PAMAM dendrimers -based non-viral gene vector, which is able to efficiently and safely transduce the Math1 gene in vitro. This vector may provide a new powerful tool of gene therapy for deafness.关键词
基因转导/聚酰胺胺树枝状聚合物/Math1/活化/环糊精修饰Key words
Gene transduction/ PAMAM dendrimers/ Math1 gene/ Activation/ Na-carboxymethyl-β -cyclodextrin modification分类
医药卫生引用本文复制引用
吴南,陈志婷,李敏,刘会占,任丽丽,吴雁,杨伟炎,杨仕明..内耳相关基础研究专辑基于聚酰胺胺的非病毒载体携带Math1基因进行体外基因转导的有效性和安全性评价[J].中华耳科学杂志,2011,9(3):258-263,6.基金项目
本课题由国家高技术研究发展计划(863计划)专题项目(2007AA022150),国家自然科学基金重点项目(30730040)和面上项目(30871398, 30571017)、国家博士后基金(第46批)联合资助. （863计划）
