An antisense approach to phenotype-based gene cloning in Tetrahymena

Proc Natl Acad Sci U S A. 2001 Jul 17;98(15):8709-13. doi: 10.1073/pnas.151243498. Epub 2001 Jul 3.


We report a pioneering approach using Tetrahymena thermophila that permits rapid identification of genes based on their null or hypomorphic phenotypes. This technique involves cell transformation with a library of plasmids that encode 26S ribosomal subunits containing short insertions. The insertions correspond to antisense sequences for a large number of genes. The majority of cells each acquires a single antisense sequence, which silences a single genomic locus. Because the insertion site within the ribosomal sequence is known, the silenced gene is easily amplified. We demonstrate that this approach can be used to identify genes required for dense core granule exocytosis.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Animals
  • Base Sequence
  • Cloning, Molecular
  • DNA, Antisense*
  • DNA, Protozoan*
  • Genes, Protozoan*
  • Molecular Sequence Data
  • Phenotype
  • Tetrahymena thermophila / genetics*


  • DNA, Antisense
  • DNA, Protozoan

Associated data

  • GENBANK/AF331706