RNA Analysis by Biosynthetic Tagging (RABT): a tool for the identification of cell type-specific RNAs

RNA Analysis by Biosynthetic Tagging (RABT): a tool for the identification of cell type-specific RNAs

Funding Type: 
Tools and Technologies I
Grant Number: 
RT1-01052
Award Value: 
$481,096
Cell Line Generation: 
Cancer Stem Cell
Status: 
Closed
Public Abstract: 
Statement of Benefit to California: 
Progress Report: 

Year 1

Our project goal is to develop and test a tool that will allow the identification of stem cell-specific genes and genes that are unique to cells made by stem cells. We have made progress in applying this technique to cells grown in culture and have made the tools necessary to identify genes expressed in specific cell types using a cell culture model of nervous system development. We have also made mice that prove this technique works in genetically-manipulated mice. These mice provide important proof-of-principle results, laying the foundation for the development of additional mouse strains that can be used to identify gene expression in stem cells and other cells of interest in their normal, in vivo, setting. We have also shown that we can selectively purify the mRNAs of genes expressed in the brain and other tissues of the mouse.

Year 2

Our project goal is to develop and test a tool that will allow the identification of stem cell-specific genes and genes that are unique to cells made by stem cells. We have optimized this technique in a model of nervous system development and demonstrated that we can selectively identify neural stem cell-specific RNA. We have also made progress in developing this technique in mice, with experiments demonstrating the feasibility of RNA purification from tissues known to have resident stem cell populations. Finally, we have generated mice that will allow us to identify stem cell-specific genes from normal stem cells at early stages of development and in adults as well as abnormal proliferating cells in mouse models of cancer.

Year 3

Stem cells exist within complex environments, surrounded by other cells. This presents a technical challenge to researchers that want to study gene expression in stem cells without removing the cells from their natural environment. We have developed a technology that makes this possible. We used the RABT technique to purify mRNAs (gene products) from stem cells of the immune system and the nervous system, in a way that was not previously possible. We have also optimized this technique for studies in various model systems, including various tissues and developmental stages in mouse models. The tools and methods we developed should be useful to scientists in many disciplines and will allow identification of genes that regulate stem cell behavior.

© 2013 California Institute for Regenerative Medicine