Project Details
Description
Abstract
Numerous recent studies have consistently shown that likely no two cells in the human body have the
same genomes, a phenomenon called somatic mosaicism. Mosaicism can be studied using various approaches,
but the study of mutations directly in the cell promises a comprehensive characterization of mosaicism in any
tissue. Analysis of single cell genome by cloning relies on natural DNA replication machinery in cells and, thus,
minimizes errors in DNA during cloning; however, cloning is limited by the ability of cells to proliferate. Analysis
by whole genome amplification (WGA) is hampered by introduced errors and non-uniformity of amplification.
Here we propose to address the limitations of single cell cloning and single cell WGA by developing a hybrid
approach that proceeds in two stages: 1) limited culturing of single cells to a micro-sized colony of 2-50 cells;
and 2) WGA of the micro-size colonies to yield enough DNA material for sequencing. An optimized hybrid
approach will enable rigorously and unbiasedly studying somatic mosaic at a single cell level throughout the
human body without WGA artifacts. Finally, to preserve tissue cell heterogeneity and enable biobanking of
tissues amenable to the developed hybrid approach, we will develop a storing protocol for tissues to preserve
proliferative potential of cells in the stored tissues. Success of the project would enable comprehensive and
accurate discovery of mutations in a single cell in a variety of tissues prioritized by SMaHT and beyond,
deepening our understanding of the mosaicism of humans.
2
Status | Finished |
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Effective start/end date | 4/15/23 → 3/31/24 |
Funding
- National Institute of Neurological Disorders and Stroke: $365,664.00
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