期刊文献+

PiggyBac转座子在肝脏中的活性研究 被引量:1

The activity of PiggyBac transposon in liver
下载PDF
导出
摘要 本实验运用高压尾静脉注射的方法将PB转座系统导入小鼠肝脏,研究其在肝脏中的转座活性,系统中PB转座子携带表达红色荧光蛋白的基因以指示转座的发生.注射10个月后取出小鼠肝脏,体视荧光显微镜观察肝脏是否有红色荧光,并通过基因组PCR、splinkerette PCR分析PB转座子在肝脏中的插入情况.实验结果表明,PB转座子在肝脏内发生高效转座,检测的68个PB插入位点中有34个位于基因序列,使得PB系统可以作为有效的基因诱变工具来研究基因功能. In this study, PB binary cotransfection system was directly delivered into liver via hydrody- namic injection using RFP (Red Fluorescence Protein) as the indicator of PB transposition. Ten months later of injection, tests were carried out to detect the red fluorescence in liver. Then, genomic PCR and splinkerette PCR were performed to analyze the insertion sites of PB transposon. The results showed that PB transposon has an efficient transposition in liver and 34/68 insertion sites were located in gene sequence.
出处 《四川大学学报(自然科学版)》 CAS CSCD 北大核心 2016年第5期1130-1134,共5页 Journal of Sichuan University(Natural Science Edition)
基金 国家自然科学基金(31300674)
关键词 PiggyBac(PB)转座子 高压尾静脉注射 Splinkerette PCR 插入突变 PiggyBac (PB) transposon Hydrodynamic injection Splinkerette PCR Insertional mutagenesis
  • 相关文献

参考文献19

  • 1Ding S, Wu X, Li G, et al. Efficient transposition of the piggyBac (PB) transposon in mammalian cells and mice I-J3. Cell, 2005, 122(3), 473.. 483.
  • 2Wang W, Lin C, Lu D, et al. Chromosomal trans- position of piggyBac in mouse embryonic stem cells [J]. Proc Natl Acad Sci USA, 2008, 105 (27), 9290: 9295.
  • 3Rad R, Rad L, Wang W, etal. PiggyBac Transpo- son Mutagenesis: A Tool for Cancer Gene Discov- ery in Mice I-J, Science, 2010, 330 (6007), 1104: 1107.
  • 4Meir Y J, Weirauch T W, Yang H S, et al. Ge- nome-wide target profiling of piggyBac and To12 in HEK 293: pros and cons for gene discovery and gene therapy [J]. BMC Biotechnology, 2011, 11: 28.
  • 5Liang Q, Kong J, Stalker J, et al. Chromosomal mobilization and reintegration of Sleeping Beauty and PiggyBac transposons [J]. Genesis, 2009, 47 (6), 404: 408.
  • 6Li M A, Pettitt S J, Eckert S, et al. The piggybac transposon displays local and distant reintegration preferences and can cause mutations at non-canonical integration sites [J]. Mol Cell Biol, 2013, 33(7), 1317: 1330.
  • 7Ley D, Van Zwieten R, Puttini S, et al. A Piggy- Bac-mediated approach for muscle gene transfer or cell therapy [J]. Stem Cell Research, 2014, 13(3)390: 403.
  • 8Matsui H, Fujimoto N, Sasakawa N, et al. Deliv- ery of full-length factor " using a piggyback tra-ns- poson vector to correct a mouse model of hemophilia A[J]. PLoSOne, 2014, 9(8), e104957.
  • 9Wilson M H, Coates C J, George A L Jr. PiggyBac transposon-mediated gene transfer in human cells [J]. Molecular Therapy, 2007, 15(1), 139145.
  • 10Di Matteo M, Samara-Kuko E, Ward N J, et al. Hyperactive PiggyBac Transposons for Sustained and Robust Liver-targeted Gene Therapy[J]. Mo- lecular Therapy, 2014, 22(9), 1614: 1624.

二级参考文献16

  • 1吴晓晖.哺乳动物PB转座因子研究[J].生命科学,2007,19(2):114-116. 被引量:5
  • 2Lander E S, Linton L M, Birren B, et al. Initial se- quencing and analysis of the human genome [J]. Na- ture. 2001; 409: 860.
  • 3McClintock B. The origin and behavior of mutable lo- ci in Maize [J]. Proc Natl Aead Sci U S A, 1950, 36 : 344.
  • 4Hutchison C A, Peterson S N, Gill S R, etal. Glob- al transposon mutagenesis and a minimal mycoplasma genome [J]. Science, 1999, 286: 2165.
  • 5Rubin G M, Spradling A C. Genetic transformation of Drosophila with transposable element vectors [J]. Science, 1982, 218: 348.
  • 6Fraser M J, Hink W F. The isolation and character- ization of MP and FP plaque variants of galleria mel- lonella nuclear polyhedrosis virus [J]. Virology, 1982, 117:366.
  • 7Fraser M J, Smith G E, Summers M D. The acquisi- tion of host cell DNA sequences by Baculoviruses: Relation between host DNA insertions and FP mu- tants of Autographa californica and Galleria mellonel- la NPVs [J]. Virology, 1983, 47: 287.
  • 8Ding S, Wu X, Li G, et al. Efficient transposition of the piggyBac (PB) transposon in mammalian cells and mice [J]. Cell, 2005, 122: 473.
  • 9Herron B J, Lu W, Rao C, et al. Efficient genera- tion and mapping of recessive developmental muta-tions using ENU mutagenesis [J]. Nature Genetics, 2002, 30: 185.
  • 10Dupuy A J, Fritz S, Largaespada D A. Transposition and gene disruption in the male germline of the mouse [J]. Genesis, 2001, 30: 82.

同被引文献5

引证文献1

二级引证文献1

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部