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腺相关病毒介导FMRP在成年Fmr1 KO小鼠脑组织中获得性表达
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Abstract:
脆性X综合征(Fragile X syndrome, FXS)是一种智力缺陷和自闭症谱系的单基因遗传疾病。位于Xq27.3染色体的Fmr1基因沉默，造成该基因编码的转录抑制因子FMRP表达缺失，从而导致FXS患者神经系统发育障碍。本研究采用质粒转染293T细胞获取AAV9-FMRP腺相关病毒载体，通过立体定位方法将AAV9-FMRP注射到成年Fmr1 KO小鼠侧脑室。免疫组化检测发现AAV9介导的FMRP可以在Fmr1 KO小鼠海马和顶叶皮层等脑组织中获得性表达；免疫荧光检测发现FMRP定位于小鼠海马神经元胞质。研究结果表明，通过腺相关病毒可以介导FMRP在成年Fmr1 KO小鼠脑组织中获得性表达。
Fragile X syndrome (FXS) is a genetic disease with mental retardation and autism spectrum, caused by the silence of Fmr1 gene on chromosome xq27.3 and the expressing loss of FMRP, which is a transcriptional inhibitor. Silencing of Fmr1 gene leads to developmental disorders of the nervous system in FXS patients. In this study, adeno-associated virus 9-mediated FMRP vector (AAV9-FMRP) was obtained by transfecting 293T cells with plasmids. AAV9-FMRP was injected into the lateral ventricle of adult Fmr1 KO mice stereotactically. Immunohistochemistry assay showed that AAV9-FMRP was able to be expressed in hippocampus and parietal cortex in Fmr1 KO mice, and immunofluorescence assay showed that FMRP expression localized in the cytoplasm of hippocampal neurons. The results indicate that it is feasible to acquire FMRP expression mediated by adeno-associated virus in brain tissues of adult Fmr1 KO mice.
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