Temporally and Spatially Restricted Gene Expression Profiling

Title:Temporally and Spatially Restricted Gene Expression Profiling

Volume: 15 Issue: 4

Author(s): Alexandra Tallafuss, Philip Washbourne and John Postlethwait

Affiliation:

Keywords: 4tU-tagging, Gene expression profiling, INTACT, RNA-seq, Transcriptome, Translatome, TRAP.

Abstract: Identifying gene function in specific cells is critical for understanding the processes that make cells unique. Several different methods are available to isolate actively transcribed RNA or actively translated RNA in specific cells at a chosen time point. Cell-specific mRNA isolation can be accomplished by the expression of transgenes in cells of interest, either directly from a specific promoter or using a modular system such as Gal4/UAS or Cre/lox. All of the methods described in this review, namely thiol-labeling of RNA (TU-tagging or RABT), TRAP (translating ribosome affinity purification) and INTACT (isolation of nuclei tagged in specific cell types), allow next generation sequencing, permitting the identification of enriched gene transcripts within the specific cell-type. We describe here the general concept of each method, include examples, evaluate possible problems related to each technique, and suggest the types of questions for which each method is best suited.

Cite this article as:

Tallafuss Alexandra, Washbourne Philip and Postlethwait John, Temporally and Spatially Restricted Gene Expression Profiling, Current Genomics 2014; 15 (4) . https://dx.doi.org/10.2174/1389202915666140602230106