| 研究実績の概要 |
Many labs around the world has been working to develop novel methods for lineage tracing employing different mechanisms. But, these methods cannot provide the information on clone size of the dys-regulated and expanded clones, which led us to design this study to develop a method to determine both clonality and clone size simultaneously.
I designed, developed and evaluated the novel clonal tracing method in cell line transfected with sleeping beauty transposon vectors and analysis of data revealed that the method can detect around 500 clones (both expanded and non-expanded) with their respective clone size. Distribution of integration site analysis showed random and stochastic distribution of transposon integration in all the chromosomes. I also evaluated the method with cell number and with as low as 10000 cells, the method work well and can detect enough clones with their respective clone size. To apply this method in vivo, novel strains of mice were needed. So, I also generated two novel strains of mice- Rosa26-Hyper sleeping beauty (HSB), which contains transposase enzyme and polyIR mice, which contains multiple copy of IR transposon in silent locus of chromosome.
Upon successful evaluation of the method developed in this study in mouse model, it will provide useful tools not only in the field of hematology but also in other biomedical research fields to trace lineage to single cell resolution. The knowledge obtained from this study is expected to provide contribution towards establishment of preventive methods for pre-leukemia initiation and progression.
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