Unknown,Transcriptomics,Genomics,Proteomics

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A highly sensitive and robust method for genome-wide 5hmC profiling of rare cell populations


ABSTRACT: Investigations of 5-hydroxymethylcytosine (5hmC) in biologically and clinically samples and models with low cell numbers have been hampered by the low sensitivity and reproducibility using current 5hmC mapping approaches. Here, we develop a selective 5hmC chemical labeling approach using tagmentation-based library preparation in order to profile nanogram levels of 5hmC isolated from ~1,000 cells (nano-hmC-Seal). Using this technology, we profiled the dynamics of 5hmC across different stages of mouse hematopoietic differentiation. Additionally, applying nano-hmC-Seal to the hematopoietic multipotent progenitor cells in an acute myeloid leukemia (AML) mouse model, we identified leukemia-specific, differentially hydroxymethylated regions that harbor previously reported and as-yet-unidentified functionally relevant factors. The change of 5hmC patterns in AML strongly correlates with the altered gene expression on a global scale. Together, our new approach offers a highly sensitive and robust method to study and detect DNA methylation dynamics from in vivo model and clinical samples. Selective 5hmC chemical labeling approach using tagmentation-based library preparation in order to profile nanogram levels of 5hmC isolated from ~1,000 cells

ORGANISM(S): Mus musculus

SUBMITTER: Dali Han 

PROVIDER: E-GEOD-77967 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Publications

A Highly Sensitive and Robust Method for Genome-wide 5hmC Profiling of Rare Cell Populations.

Han Dali D   Lu Xingyu X   Shih Alan H AH   Nie Ji J   You Qiancheng Q   Xu Meng Michelle MM   Melnick Ari M AM   Levine Ross L RL   He Chuan C  

Molecular cell 20160728 4


We present a highly sensitive and selective chemical labeling and capture approach for genome-wide profiling of 5-hydroxylmethylcytosine (5hmC) using DNA isolated from ∼1,000 cells (nano-hmC-Seal). Using this technology, we assessed 5hmC occupancy and dynamics across different stages of hematopoietic differentiation. Nano-hmC-Seal profiling of purified Tet2-mutant acute myeloid leukemia (AML) murine stem cells allowed us to identify leukemia-specific, differentially hydroxymethylated regions tha  ...[more]

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