An Efficient Transgenesis Approach for Gene Delivery in the Mouse Embryonic Heart

被引:0
|
作者
Manes-Garcia, Jorge [1 ]
Beccari, Leonardo [1 ]
Torres, Miguel [2 ,3 ]
Ocana, Oscar H. [2 ,4 ]
机构
[1] Severo Ochoa Ctr Mol Biol CBMSO, Tissue & Organ Homeostasis Program, Madrid, Spain
[2] Natl Cardiovasc Res Ctr CN, Cardiovasc Regenerat Program, Houston, TX 77004 USA
[3] Ctr Invest Biomed Red Enfermedades Cardiovasc CIBE, Madrid, Spain
[4] Univ Jaen, Fac Expt Sci, Dept Expt Biol, Jaen, Spain
来源
基金
欧盟地平线“2020”;
关键词
D O I
10.3791/66754
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
The mammalian heart is a complex organ formed during development via highly diverse populations of progenitor cells. The origin, timing of recruitment, and fate of these progenitors are vital for the proper development of this organ. The molecular mechanisms that govern the morphogenesis of the heart are essential for understanding the pathogenesis of congenital heart diseases and embryonic cardiac regeneration. Classical approaches to investigate these mechanisms employed the generation of transgenic mice to assess the function of specific genes during cardiac development. However, mouse transgenesis is a complex, time-consuming process that often cannot be performed to assess the role of specific genes during heart development. To address this, we have developed a protocol for efficient electroporation and culture of mouse embryonic hearts, enabling transient transgenesis to rapidly assess the effect of gain- or loss-of-function of genes involved in cardiac development. Using this methodology, we successfully overexpressed Meis1 in the embryonic heart, with a preference for epicardial cell transfection, demonstrating the capabilities of the technique.
引用
收藏
页数:12
相关论文
共 50 条
  • [1] Efficient clonal gene delivery into human embryonic stem cells via active transgenesis using piggyBac transposition
    Zhang, Ting-Ting
    Ulrey, Clayton L.
    Liu, Shan-Run
    Yang, Rui
    McConnell, Sean C.
    Huo, Yongliang
    Ryan, Thomas M.
    [J]. BLOOD, 2007, 110 (11) : 369B - 369B
  • [2] Avian embryonic stem cells: a new approach for efficient avian transgenesis.
    Pain, B
    Risson, V
    Samarut, J
    [J]. EUROPEAN JOURNAL OF CELL BIOLOGY, 1997, 74 : 121 - 121
  • [3] Efficient gene targeting in mouse embryonic stem cells
    Templeton, NS
    Roberts, DD
    Safer, B
    [J]. GENE THERAPY, 1997, 4 (07) : 700 - 709
  • [4] Efficient gene targeting in mouse embryonic stem cells
    NS Templeton
    DD Roberts
    B Safer
    [J]. Gene Therapy, 1997, 4 : 700 - 709
  • [5] Cationic star copolymers based on β-cyclodextrins for efficient gene delivery to mouse embryonic stem cell colonies
    Loh, Xian Jun
    Wu, Yun-Long
    [J]. CHEMICAL COMMUNICATIONS, 2015, 51 (54) : 10815 - 10818
  • [6] Mouse Astrocytoma Models: Embryonic Stem Cell Mediated Transgenesis
    Hao Ding
    Abhijit Guha
    [J]. Journal of Neuro-Oncology, 2001, 53 : 289 - 296
  • [7] Mouse astrocytoma models: Embryonic stem cell mediated transgenesis
    Ding, H
    Guha, A
    [J]. JOURNAL OF NEURO-ONCOLOGY, 2001, 53 (03) : 289 - 296
  • [8] Efficient gene delivery in differentiated human embryonic stem cells
    Kim, JH
    Do, HJ
    Choi, SJ
    Choi, HJ
    Park, KH
    Yang, HM
    Lee, SH
    Kim, DK
    Kwack, K
    Oh, SK
    Moon, SY
    Cha, KY
    Chung, HM
    [J]. EXPERIMENTAL AND MOLECULAR MEDICINE, 2005, 37 (01): : 36 - 44
  • [9] Efficient gene delivery in differentiated human embryonic stem cells
    Kim J.-H.
    Do H.-J.
    Choi S.-J.
    Cho H.-J.
    Park K.-H.
    Yang H.-M.
    Lee S.-H.
    Kim D.-K.
    Kwack K.
    Oh S.-K.
    Moon S.-Y.
    Cha K.-Y.
    Chung H.-M.
    [J]. Experimental & Molecular Medicine, 2005, 37 (1) : 36 - 44
  • [10] Gene-manipulated embryonic stem cells for rat transgenesis
    Kawamata, Masaki
    Ochiya, Takahiro
    [J]. CELLULAR AND MOLECULAR LIFE SCIENCES, 2011, 68 (11) : 1911 - 1915