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  • ISSN 1006-3080
  • CN 31-1691/TQ

靶向心肌组织的新型腺相关病毒载体的构建及筛选

陈晨 杜增民 吴侠 蒋威 赵阳 肖啸 郑静

陈晨, 杜增民, 吴侠, 蒋威, 赵阳, 肖啸, 郑静. 靶向心肌组织的新型腺相关病毒载体的构建及筛选[J]. 华东理工大学学报(自然科学版), 2021, 47(3): 284-291. doi: 10.14135/j.cnki.1006-3080.20200313002
引用本文: 陈晨, 杜增民, 吴侠, 蒋威, 赵阳, 肖啸, 郑静. 靶向心肌组织的新型腺相关病毒载体的构建及筛选[J]. 华东理工大学学报(自然科学版), 2021, 47(3): 284-291. doi: 10.14135/j.cnki.1006-3080.20200313002
CHEN Chen, DU Zengmin, WU Xia, JIANG Wei, ZHAO Yang, XIAO Xiao, ZHENG Jing. Construction and Screening of A Novel Adeno-Associated Virus Vector Targeting Myocardial Tissue[J]. Journal of East China University of Science and Technology, 2021, 47(3): 284-291. doi: 10.14135/j.cnki.1006-3080.20200313002
Citation: CHEN Chen, DU Zengmin, WU Xia, JIANG Wei, ZHAO Yang, XIAO Xiao, ZHENG Jing. Construction and Screening of A Novel Adeno-Associated Virus Vector Targeting Myocardial Tissue[J]. Journal of East China University of Science and Technology, 2021, 47(3): 284-291. doi: 10.14135/j.cnki.1006-3080.20200313002

靶向心肌组织的新型腺相关病毒载体的构建及筛选

doi: 10.14135/j.cnki.1006-3080.20200313002
基金项目: 中央高校基本科研业务费专项资金资助
详细信息
    作者简介:

    陈晨:陈 晨(1994-),男,甘肃省兰州市人,硕士生,主要研究方向为基因治疗。E-mail:cchenecust@163.com

    通讯作者:

    郑 静,E-mail:zhengjing@ecust.edu.cn

  • 中图分类号: Q819

Construction and Screening of A Novel Adeno-Associated Virus Vector Targeting Myocardial Tissue

  • 摘要: 为了构建以心肌为靶点的基因治疗载体,通过DNA shuffling技术对选择的8种AAV(AAV1~AAV4,AAV6~AAV9)衣壳基因进行了重组,构建了随机突变体AAV衣壳文库。将随机突变体AAV衣壳文库在小鼠体内直接筛选,最终筛选得到高心肌靶向和低肝脏靶向的新型AAV衣壳:AAVH50与AAVH59。通过尾静脉注射,将AAV携带的荧光素酶(Luc)基因递送到小鼠体内,结果表明:在检测的11个组织中,AAVH50与AAVH59在心肌组织的基因表达高于其他组织。与AAV6、AAV9相比,AAVH50与AAVH59心肌中的Luc表达水平相近,肝脏中的表达明显低于AAV9。AAVH50和AAVH59感染原代新生大鼠心肌细胞后,AAVH59在心肌直接感染的效率明显高于AAV9。

     

  • 图  1  DNA shuffling技术构建突变体AAV衣壳文库与小鼠体内筛选

    Figure  1.  Construction of mutant AAV capsid library by DNA shuffling technology and in vivo screening in mice

    图  2  AAVH50与AAVH59的衣壳序列分析

    Figure  2.  Sequence analysis of the AAVH50与AAVH59 capsid

    图  3  注射AAV9-CMV-Luc、AAVH50-CMV-Luc、AAVH59-CMV-Luc后小鼠荧光素酶活体成像。

    Figure  3.  In vivo imaging of mouse luciferase after injection of AAV9-CMV-Luc、AAVH50-CMV-Luc、AAVH59-CMV-Luc

    图  4  尾静脉注射AAV载体后各种小鼠组织中的荧光素酶活性

    Figure  4.  Luciferase activity in various mouse tissues after tail vein injection of AAV vector

    图  5  尾静脉注射AAV载体后各种小鼠组织中载体基因组拷贝数

    Figure  5.  Vector genome copy numbers in various mouse tissues after tail vein injection of AAV vector

    图  6  不同AAV载体在原代心肌细胞上的转导效率

    Figure  6.  Transduction efficiency of different AAV vectors on primary cardiomyocytes

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出版历程
  • 收稿日期:  2020-03-13
  • 网络出版日期:  2020-07-22
  • 刊出日期:  2021-06-30

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