Molecular analysis of single neural progenitor cells during brain development

Funding Type: 
Tools and Technologies I
Grant Number: 
ICOC Funds Committed: 
Public Abstract: 
With the complete sequencing of the human genome, the current challenge is to learn how the products of the 30,000 – 150,000 identified genes interact to produce the complexed human body, which contains very diverse cell types with diverse functions, and how diseases arise when either the functions of these genes or their regulations go awry. To meet this challenge, we need to know what genes are being expressed at a particular time in a particular cell. Currently technology is being developed to study the expression of many genes at once. However these technologies are limited to study gene expression in millions, or thousands of cells, thereby restricting the study of small samples or complex tissues, especially cells that are very rare in the body, such as the stem cells. To understand how genes regulate stem cells maintaining their “stemness” and differentiation, we propose a strategy to study gene expression in individual cells by sequencing all the transcripts in a single neural stem cell taken out from a developing mouse embryo, by combining two latest technologies – Connector Inversion Probe (CIP) technology and the SOLiD Sequencing platform from Applied Biosystems, which is one of the latest DNA sequencing technology that has a capacity of 114 million sequencing reactions per run. The CIP technology allows the rapid cloning and amplification of all the transcripts in the cell. In the current CIRM proposal, we propose to further develop the CIP technology, and to test our strategy to sequence all the transcripts in a single cell, we propose to sequence 1000 genes in individual neural stem cells. If this strategy proves successful, it will provide deep understanding how many human diseases arise at a single cell level, and facilitate scientists and physicians to develop novel and effective therapeutic treatments.
Statement of Benefit to California: 
Almost all human illness, perhaps except trauma, have some genetic basis. While the complete human genome is sequenced, it is still not clear how mutation in gene function or alteration in their regulation leads to human diseases. In this CIRM proposal, we propose to develop a strategy to sequence all the transcript in a single cell. If this strategy is successful, it will enable scienctists and physicians to have a deep understanding how certain human diseases arise and progress at a single cell level. It will guide the development of novel diagnosis and/or treatment strategies. Everyone, include the citizens of California, will benefit tremendously from this research.

© 2013 California Institute for Regenerative Medicine