Unit

UNIT 9.5 Selection of Transfected Mammalian Cells

  1. Richard M. Mortensen1,
  2. Robert E. Kingston2

Published Online: 1 APR 2009

DOI: 10.1002/0471142727.mb0905s86

Current Protocols in Molecular Biology

Current Protocols in Molecular Biology

How to Cite

Mortensen, R. M. and Kingston, R. E. 2009. Selection of Transfected Mammalian Cells. Current Protocols in Molecular Biology. 86:I:9.5:9.5.1–9.5.13.

Author Information

  1. 1

    University of Michigan, Ann Arbor, Michigan

  2. 2

    Massachusetts General Hospital and Harvard Medical School, Boston, Massachusetts

Publication History

  1. Published Online: 1 APR 2009
  2. Published Print: APR 2009

Abstract

To determine the function of a gene in vitro, expression in heterologous cells is often employed. This can be done by transient expression, but often requires a more permanent expression of the gene and the creation of a cell line. This process can involve decisions as to the nature of construct used for expression, and invariably uses some strategy to select the transfected cells. Typically, these strategies use one of a number of genes that confer resistance to an added drug that will kill untransfected cells but not the transfected cells (positive selection). Alternatively, sometimes the strategy uses a gene that will confer sensitivity to a compound and kills the transfected cells (negative selection). This chapter discusses some of the strategies and genes used in creating cell line for in vitro study of gene function. Curr. Protoc. Mol. Biol. 86:9.5.1-9.5.13. © 2009 by John Wiley & Sons, Inc.

Keywords:

  • stable integration;
  • selection;
  • mammalian cell;
  • stable transfection;
  • selection marker;
  • Cre-lox;
  • Flp-FRT;
  • expression