Abstract
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.
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Notes
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Sebastien Chenuet and Madiha Derouazi contributed equally to this work.
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Chenuet, S., Derouazi, M., Hacker, D., Wurm, F. (2009). DNA Delivery by Microinjection for the Generation of Recombinant Mammalian Cell Lines. In: Carroll, D. (eds) Microinjection. Methods in Molecular Biology, vol 518. Humana Press. https://doi.org/10.1007/978-1-59745-202-1_8
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DOI: https://doi.org/10.1007/978-1-59745-202-1_8
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