Li Ru-Shu, Yu Dan, Luo Bao-Zheng, Bo Qing-Ru, Xu Hai-Nie, Sha Cai-Hua, Liao Xiu-Yun
Zhuhai Epidemic Prevention and Supervision Center for Animals and Plants, Zhuhai 519015, China.
Bing Du Xue Bao. 2013 Jun;29(4):386-91.
In order to study the proliferation inhibition effect of H5N1 subtype avian influenza virus (AIV) with small interfere RNA (siRNA), a total of 4 siRNAs were designed in accordance with the NP and PA genes of H5N1 subtype AIV, the siRNAs were then transfected to chicken embryo fibroblast(CEF), CEF was infected with H5N1 subtype AIV after 6 hrs. Virus titer of cell supernatant was tested at 16-56hrs post infection, and pathological changes of the cells was observed; mRNA levels of NP, PA, HA and p13-actin gene were tested at 36hrs post infection. The results showed that these 4 siRNAs could inhibit the prolif-eration of H5N1 subtype AIV in CEF in varying degrees, and one siRNA targeting PA was best per-formed. The experimental results also showed that the inhibition effect was decreased with the time prolonged. This research provides a basis for further studying RNAi on AIV prevention and control.
为研究小干扰RNA（siRNA）对H5N1亚型禽流感病毒（AIV）的增殖抑制作用，根据H5N1亚型AIV的NP和PA基因设计了4条siRNA，将其转染至鸡胚成纤维细胞（CEF），6小时后用H5N1亚型AIV感染CEF。感染后16 - 56小时检测细胞上清液的病毒滴度，观察细胞的病理变化；感染后36小时检测NP、PA、HA和β-actin基因的mRNA水平。结果显示，这4条siRNA均可不同程度抑制H5N1亚型AIV在CEF中的增殖，其中靶向PA的1条siRNA效果最佳。实验结果还显示，随着时间延长抑制效果下降。本研究为进一步研究RNA干扰技术防控AIV提供了依据。
