Several tools for genome-wide expression studies have been developed in the recent years, which led to the identification of many plant genes and proteins involved in different developmental and physiological processes. Yet, the specificity of information derived from RNA and protein expression profiling is limited to the specificity of the biological starting material. Higher plants consists of several distinct tissues and cell types, each contributing to the overall performance of the whole organism. Most DNA microarray expression analyses and proteome studies have been performed on whole organs or tissues, thus only providing average measurements which mask differences between different cell types. This averaging effect can only be overcome by increasing the spatial resolution to a cell-specific level. Methods such as immunolocalization, in situ hybridization, and reporter gene expression provide the spatial resolution needed to investigate gene expression or protein accumulation in single cells, but they only allow the analysis of individual genes or proteins. Alternative approaches, as the Laser Capture Microdissection (LCM) technique and gene trapping systems, can provide such cellular information on a genome-wide scale. LCM is a rapid method of isolating pure cellular preparations directly from heterogeneous tissues, based on conventional histological identification. LCM-harvested cells can provide DNA, RNA, or protein for the profiling of genomic characteristics, gene expression and protein spectra from individual cell types. Gene trap lines, in which endogenous proteins are fused to reporter markers, visually reveal the pattern of gene expression within individual cells in complex tissues. Most importantly, gene trap insertions provide information on gene function, as they often disrupt the tagged gene and give rise to mutant phenotypes.
|Titolo:||From complex tissues to single cell: genomic approaches for the identification of cell-specific genes|
|Settore Scientifico Disciplinare:||Settore BIO/18 - Genetica|
Settore INF/01 - Informatica
|Data di pubblicazione:||2008|
|Enti collegati al convegno:||Società Italiana di Fisiologia Vegetale|
|Tipologia:||Book Part (author)|
|Appare nelle tipologie:||03 - Contributo in volume|