滴度及注射方式对2型腺相关病毒载体表达效果的影响

由于腺相关病毒载体(adeno-associated viral vector,AAV)本身的侵染不具有神经元特异性,其在神经系统相关研究中会存在侵染范围不符合试验要求的情况,因此本试验拟研究不同滴度和注射方式对病毒侵染范围的影响。结果显示,在注射较高滴度的AAV2-CMV-EGFP(1.3×10¹¹mg/L)3周后,其侵染范围可由延髓中缝苍白核(raphe pallidus nucleus,RPa)区域延伸至下丘脑室旁核(paraventricular nucleus,PVN)区域;较低滴度(1.3×10⁸mg/L)结合压力注射的方式可以在一定程度上限制AAV的侵染范围,并且载体携带的EGFP荧光基团在637 nm波段不具有明显的假阳性信号。

Effects of viral dose and injection method on expression of adeno-associated viral vector serotype 2

In recent years,adeno-associated viral(AAV)has been widely used as a new and efficient research tool in the life science research.Because infection of adenovirus vector itself is not neuron-specific,there may be cases where the range of AAV infection does not meet the requirements of the study in nervous system.Therefore,this experiment intends to investigate the effect of different dosages of virus and injection methods on infection of the virus vector.The results showed that,after being injected with high dosage of AAV2-CMV-EGFP(1.3×10¹¹mg/L)for three weeks,the infection range could extend from the area of the raphe pallidus nucleus(RPa)in medulla to the paraventricular nucleus(PVN)in hypothalamus,whereas low dosage of vector(1.3×10⁸mg/L)combined with pressure injection method can limit the range of AAV infection,and the EGFP carried by the vector has no obvious false positive signal at 637 nm.