Designation and silencing function of the small interfering RNA of HLA-A2

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Abstract

BACKGROUND:

Human leukocyte antigen (HLA), the major histocompatibility complex of human, plays an important function in the transplant rejection. Decreasing the expression of HLA will prolong the survival time of transplants.

OBJECTIVE:

To design small interfering RNA (siRNA) of HLA-A2 and to detect the effect of siRNA-HLA-A2 on the expression of HLA-A2.

METHODS:

Four kinds of siRNA-HLA-A2 domains were designed, and recombinant lentivirus expression vector were formed. The 293T cells, highly expressing HLA-A2, were infected in vitro. Then the knockout efficacy of four domains was detected to select the highly efficient siRNA-HLA-A2 target sequences. The human embryo lung fibroblasts were cultured in vitro and infected with the lentivirus carrying the target sequence. The infecting efficiency of LV-siRNA-HLA-A2 was observed under the fluorescence microscope and the silence function of this siRNA in human embryo lung fibroblasts was detected by western blot analysis.

RESULTS AND CONCLUSION:

According to the mRNA sequence of HLA-A2 in Genbank, three siRNAs were designed and synthesized. In vitro, the over expression of HLA-A2 in 293K cells was successfully silenced. The HLA-A2 expression in human embryo lung fibroblasts was also efficiently silenced after the human embryo lung fibroblasts were infected by the highly efficient siRNA of HLA-A2. The efficacy was up to 80%.

Subject headings:

HLA antigens; RNA interference; gene silencing; fibroblasts

Funding:

the Natural Science Foundation of Liaoning Province, No. 2013022066, 2013022048; a grant by Liaoning Provincial Education Bureau, No. L2013335; the Youth Fund of Liaoning Medical University, No. Y2011Z022

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