腺相关病毒载体的杆状病毒生产系统的构建  

Establishment of Production System of Recombinant Adeno-associated Viruses Based on Baculoviruses

作　　者：雷求刚[1] 徐增辉[2] 钱其军[1,2] 

机构地区：[1]浙江理工大学生命科学学院,杭州310018 [2]第二军医大学东方肝胆外科医院,上海200438

出　　处：《浙江理工大学学报（自然科学版）》2010年第5期773-778,共6页Journal of Zhejiang Sci-Tech University(Natural Sciences)

基　　金：国家自然科学基金(30772477)

摘　　要：由于腺相关病毒(adeno-associated virus,AAV)的非致病性及能感染多种组织类型,重组AAV载体(rAAV)在基因治疗领域备受关注。然而,如何高效生产rAAV以适应临床上对其高产量的需求一直是限制其应用的瓶颈。借助杆状病毒携带rAAV包装所需组分,通过感染昆虫细胞来生产rAAV是一种能够解决其产量瓶颈的生产方式。将AAV2的Rep、Cap基因序列以及含AAV2 ITR及EGFP表达框的序列分别构建到三个杆状病毒中,通过共感染Sf9细胞来生产rAAV,结果显示成功包装出rAAV病毒,且病毒活力较好。这种生产方式很容易放大,将为AAV载体产业化及临床应用奠定基础。Adeno-associated virus is nonpathogenic to the human body and can infect multiple tissue types.For these characteristic features,AAV vectors are being given more and more attention.However,the major obstacle to broaden the use of rAAV vectors remains the limited capacity of available production systems to provide sufficient rAAV quantities for clinical trials.Using baculovirus to carry the elements necessary for AAV package to produce rAAV in insect cells is an ideal production method.The authors applies three baculoviruses carrying AAV2 Rep,Cap and ITR-EGFP sequences to produce rAAV in Sf9 cells.The results prove that rAAVs are successfully produced and shows good activity.This production method can be easilily scaled up,which will lay a good foundation for the industrialization and clinical application of AAV vectors.

关 键 词：RAAV 杆状病毒 生产系统 

分 类 号：Q789[生物学—分子生物学]