General Characteristics of Microbubble-Adenovirus Vectors Carrying Genes
General Characteristics of Microbubble-Adenovirus Vectors Carrying Genes
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携带基因的微泡腺病毒载体的一般特征
DOI：
10.1007/s12195-020-00663-8
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发表时间：
2021-04-01
影响因子：
2.8
通讯作者：
Mu, Yuming
中科院分区：
文献类型：
--
作者：
Yang, Lingjie;Ma, Juan;Mu, Yuming
基因导入--安全地转移基因、靶向细胞、实现高效的基因转导是基因治疗中亟待解决的问题。将微泡(MBS)与病毒结合构建双载体已成为一种很有前途的基因传递方法。了解携带基因的MBS的特征性能是促进有效基因转移的关键。方法：构建携带趋化因子(C-X-C基序)配体12(CXCL12)和骨形态发生蛋白-2(BMP2)基因(分别为pAD-CXCL12和pAD-BMP2)的MB-腺病毒载体，以探索携带基因的双载体的一般特性。结果：MB-腺病毒载体具有稳定的物理性质，其直径、浓度和pH与裸MBS(p>；0.05)无显著差异。用流式细胞仪和RT-PCR检测MBS的基因载量。MBS的基因载药率随着病毒量的增加而增加，当加入10.0亩L病毒时，载药率最高(91%)。超过10.0亩的L添加病毒后，MBS的基因载量效率随着病毒的不断添加而降低。百亩L MBS中pAD-CXCL12和pAD-BMP2的最大载量分别约为14和10亩L。结论病毒载量的增加会导致MB载量降低，MBS最大载药量可能因病毒携带的基因不同而不同。
Introduction-Transferring genes safely, targeting cells and achieving efficient transfection are urgent problems in gene therapy that need to be solved. Combining microbubbles (MBs) and viruses to construct double vectors has become a promising approach for gene delivery. Understanding the characteristic performance of MBs that carry genes is key to promoting effective gene transfer. Therefore, in this study, we constructed MB-adenovirus vectors and discussed their general characteristics.Methods-We constructed MB-adenovirus vectors carrying the chemokine (C-X-C motif) ligand 12 (Cxcl12) and bone morphogenetic protein-2 (Bmp2) genes (pAd-Cxcl12 and pAd-Bmp2, respectively) to explore the general characteristics of double vectors carrying genes.Results-The MB-adenovirus vectors had stable physical properties, and no significant differences in diameter, concentration, or pH were noted compared with naked MBs (p > 0.05). Flow cytometry and RT-PCR were used to detect the gene-loading capacity of MBs. The gene-loading efficiency of MBs increased with increasing virus amounts and was highest (91%) when 10.0 mu L of virus was added. Beyond 10.0 mu L of added virus, the gene-loading efficiency of MBs decreased with the continuous addition of virus. The maximum amounts of pAd-Cxcl12 and pAd-Bmp2 in 100 mu L of MBs were approximately 14 and 10 mu L, respectively.ConclusionsThis study indicates that addition of an inappropriate viral load will result in low MB loading efficiency, and the maximum amount of genes loaded by MBs may differ based on the genes carried by the virus.