Microinjection pp 99-112

Part of the Methods in Molecular Biology book series (MIMB, volume 518) | Cite as

DNA Delivery by Microinjection for the Generation of Recombinant Mammalian Cell Lines

  • Sebastien Chenuet
  • Madiha Derouazi
  • David Hacker
  • Florian Wurm


Gene transfer methods for producing recombinant cell lines are often not very efficient. One reason is that the recombinant DNA is delivered into the cell cytoplasm and only a small fraction reaches the nucleus. This chapter describes a method for microinjecting DNA directly into the nucleus. Direct injection has several advantages including the ability to deliver a defined copy number into the nucleus, the avoidance of DNAses that are present in the cell cytoplasm, and the lack of a need for extensive subcloning to find the recombinant cells. The procedure is described for two cell lines, CHO DG44 and BHK-21, using green fluorescent protein as a reporter gene. However, this method could easily be adapted to other cells lines and using other recombinant genes.

Key words

Recombinant cell line microinjection green fluorescent protein transfection gene expression 


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Copyright information

© Humana Press, a part of Springer Science+Business Media, LLC 2009

Authors and Affiliations

  • Sebastien Chenuet
    • 1
  • Madiha Derouazi
    • 1
  • David Hacker
    • 1
  • Florian Wurm
    • 1
  1. 1.École Polytechnique Féderale de LausanneEPFL-SV-IBI-LBTCLausanneSwitzerland

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