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 µL of virus was added. Beyond 10.0 µL 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 µL of MBs were approximately 14 and 10 µL, respectively.

Conclusions

This 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.

中文翻译：

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携带基因的微泡-腺病毒载体的一般特征

介绍

安全转移基因、靶向细胞、实现高效转染是基因治疗中亟待解决的问题。结合微泡（MB）和病毒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 MB 的特征性能是促进有效基因转移的关键。因此，在本研究中，我们构建了 MB 腺病毒载体并讨论了它们的一般特征。

方法

我们构建了携带趋化因子（CXC 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征。

结果

MB-腺病毒载体具有稳定的物理性质，与裸MB相比，其直径、浓度或pH值均无显着差异( p > 0.05)。流式细胞术和RT-PCR用于检测MBs的基因负载能力。MBs 的基因加载效率随着病毒量的增加而增加，当添加 10.0 µL 病毒时最高 (91%  )。添加病毒超过 10.0 µL， MBs的基因加载效率随着病毒的持续添加而降低。100 µL MB 中 pAd -Cxcl12和 pAd -Bmp2的最大量分别约为 14 和 10 µL。

结论

该研究表明，添加不适当的病毒载量会导致 MB 加载效率低，并且 MB 加载的最大基因量可能因病毒携带的基因而异。