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2009, Number 1

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Vet Mex 2009; 40 (1)

Lipofection improves gene targeting efficiency in E14 TG2a mouse embryonic stem cells

López-Heydeck SM, Cajero-Juárez M, Alonso-Morales RA, Martínez-Castañeda JS, Robles-González JF, Barbabosa-Pliego A, Vázquez-Chagoyán JC

Language: English/Spanish
References: 23
Page: 85-93
PDF size: 343.83 Kb.


ABSTRACT

Electroporation has been the method of election for transfection of murine embryonic stem cells for over 15 years; however, it is a time consuming protocol because it requires large amounts of DNA and cells, as well as expensive and delicate equipment. Lipofection is a transfection method that requires lower amounts of cells and DNA than electroporation, and has proven to be efficient in a large number of cell lines. It has been shown that after lipofection, mouse embryonic stem cells remain pluripotent, capable of forming germ line chimeras and can be transfected with greater efficiency than with electroporation; however, gene targeting of mouse embryonic stem cells by lipofection has not been reported. The objective of this work was to fi nd out if lipofection can be used as efficiently as electroporation for regular gene targeting protocols. This context compares gene targeting efficiency between these techniques in mouse embryonic stem cells E14TG2a, using a gene replacement type vector. No differences were found in gene targeting efficiency between groups; however, lipofection was three times more efficient than electroporation in transfection efficiency, which makes lipofection a less expensive alternative method to produce gene targeting in mouse embryonic stem cells.


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