基础医学与临床 ›› 2017, Vol. 37 ›› Issue (11): 1513-1518.

• 研究论文 • 上一篇    下一篇

长链非编码RNA AK046999在小鼠大脑皮质发育中的表达及功能

李嘉宁1,周俊洁1,刘伟1,彭小忠2,舒鹏程3   

  1. 1. 中国医学科学院基础医学研究所
    2. 中国医学科学院 基础医学研究所 北京协和医学院 基础学院 医学分子生物学国家重点实验室
    3. 中国医学科学院 基础医学研究所
  • 收稿日期:2017-07-06 修回日期:2017-09-16 出版日期:2017-11-05 发布日期:2017-11-01
  • 通讯作者: 舒鹏程 E-mail:schubert1122@163.com
  • 基金资助:
    国家重点研发计划;重点基础研究发展计划;国家自然科学基金;医科院创新基金

Expression and function of long non-coding RNA AK046999 in the development of mouse cerebral cortex

  • Received:2017-07-06 Revised:2017-09-16 Online:2017-11-05 Published:2017-11-01
  • Contact: Peng-cheng SHU E-mail:schubert1122@163.com

摘要: 目的 检测长链非编码RNA AK046999在小鼠大脑皮质发育过程中的表达及作用。方法 1)通过UCSC基因组浏览器获取其基因组座位,利用Coding Potential Calculator工具对其编码能力进行预测;2)原位杂交及免疫荧光技术确定其在大脑皮质发育过程中的表达谱;3)TALEN技术构建AK046999完全敲除小鼠,并且在DNA及RNA水平对其进行敲除鉴定;4)免疫荧光及TUNNEL技术对敲除小鼠大脑皮质在发育过程的功能进行检测。结果 1)AK046999可认为是非编码RNA(Coding Potential Score<-1);2)AK046999在小鼠大脑皮质发育过程中,前体区相对高表达;3)成功构建了AK046999敲除小鼠;4)与对照相比,AK046999敲除小鼠中表达PAX6、TBR2和NEUROD2的细胞数量无明显改变,并且在这两组小鼠中凋亡细胞的数量无明显改变。结论 AK046999在发育期的小鼠大脑皮质表达,但敲除后不影响小鼠大脑皮质中神经祖细胞的增殖、分化和细胞凋亡过程。

关键词: AK046999, 长链非编码RNA, 时空表达谱, 大脑皮层发育

Abstract: Objective To detect the expression and the role of the long non-coding RNA AK046999 in the development of mouse cerebral cortex. Methods 1) The AK046999 locus was obtained from the UCSC genome browser, and the coding ability of protein was predicted by Coding Potential Calculator; 2) The expression profile of AK046999 in the development of mouse cerebral cortex was analyzed by immunofluorescence and in situ hybridization ; 3) The AK046999 knockout mouse was constructed by TALEN technique and identified at the level of DNA and RNA; 4)The phenotype of knockout mouse cerebral cortex was analyzed by immunofluorescence and TUNNEL assay. Results 1) AK046999 had weak coding ability and could be considered as non-coding RNA; 2) AK046999 was highly expressed in VZ/SVZ of the development of mouse cerebral cortex; 3) The AK046999 knockout mouse was successfully constructed; 4) Immunofluorescence results showed that there were no obvious change in the markers of PAX6, TBR2 and NEUROD2 compared with the control, and apoptotic cells also had no obvious change. Conclusions The AK046999 is expressed in the cerebral cortex of developing mouse and AK046999 knockout does not affect the proliferation, differentiation and apoptosis of neural progenitor cells in the cerebral cortex.

Key words: AK046999, long non-coding RNA, spatio-temporal expression, cerebral cortex development

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