A versatile in vivo system for directed dissection of gene expression patterns

Nat Methods. 2011 Mar;8(3):231-7. doi: 10.1038/nmeth.1561.

Abstract

Tissue-specific gene expression using the upstream activating sequence (UAS)–GAL4 binary system has facilitated genetic dissection of many biological processes in Drosophila melanogaster. Refining GAL4 expression patterns or independently manipulating multiple cell populations using additional binary systems are common experimental goals. To simplify these processes, we developed a convertible genetic platform, the integrase swappable in vivo targeting element (InSITE) system. This approach allows GAL4 to be replaced with any other sequence, placing different genetic effectors under the control of the same regulatory elements. Using InSITE, GAL4 can be replaced with LexA or QF, allowing an expression pattern to be repurposed. GAL4 can also be replaced with GAL80 or split-GAL4 hemi-drivers, allowing intersectional approaches to refine expression patterns. The exchanges occur through efficient in vivo manipulations, making it possible to generate many swaps in parallel. This system is modular, allowing future genetic tools to be easily incorporated into the existing framework.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Bacterial Proteins / genetics
  • Base Sequence
  • DNA-Binding Proteins / genetics
  • Drosophila melanogaster / genetics*
  • Enhancer Elements, Genetic
  • Gene Expression Profiling / methods*
  • Gene Expression*
  • Molecular Sequence Data
  • Recombination, Genetic
  • Repressor Proteins / genetics
  • Saccharomyces cerevisiae Proteins / genetics
  • Serine Endopeptidases / genetics
  • Transcription Factors / genetics

Substances

  • Bacterial Proteins
  • DNA-Binding Proteins
  • GAL4 protein, S cerevisiae
  • GAL80 protein, S cerevisiae
  • LexA protein, Bacteria
  • Repressor Proteins
  • Saccharomyces cerevisiae Proteins
  • Transcription Factors
  • Serine Endopeptidases

Associated data

  • GENBANK/HQ888842
  • GENBANK/HQ888843
  • GENBANK/HQ888844
  • GENBANK/HQ888845
  • GENBANK/HQ888846