Technology

 

Single-cell itChIP

Single-cell simultaneous indexing and tagmentation-based ChIP-seq.

itChIP-seq is a low-cost and efficient method, compatible with both low cellular input and single cells for profiling chromatin states. itChIP combines chromatin opening, simultaneous cellular indexing and chromatin tagmentation within a single tube, enabling the processing of samples from tens of single cells to, more commonly, thousands of single cells per assay. We demonstrate that single-cell itChIP-seq (sc-itChIP- seq) yields ~9,000 unique reads per cell and is widely applicable and affordable for most biomedical research laboratories. (Ai et al. Nature Cell Biology, 2019)

CoBATCH

Combinatorial barcoding and targeted chromatin release.

CoBATCH is a high-throughput single-cell ChIP-seq technology. Based on fusion protein-protein A-Tn5 (PAT), we develop in situ ChIP, which is suitable for as low as 100 initial cells and intact tissues. PAT is enriched through specific antibodies to genomic regions, and Tn5 generates indexed chromatin ready for library preparation and sequencing. By introducing combinatorial indexing strategy, we develop CoBATCH, a high efficient and high-throughput single-cell histone modification and DNA-binding protein profiling method, suitable for both native and fixed cell condition. CoBATCH produces ∼12,000 non-duplicated reads per cell with extremely low background. (Wang et al. Molecular Cell, 2019)

 

CoTECH

A joint method for profiling transcriptome and chromatin occupancy simutanously in single-cells.

We intergrated our previously developed CoBATCH together with modified single-cell RNA-seq method, to simutanously profile gene expression and chromatin binding in the same single cells. Furthermore, by aggregating data from cells with similar transcriptome (data from different experimental batches), we can obtain “pseudosingle-cells” that contain informations of several histone modifications or TFs binding, enabling higher-dimensional epigenomic reconstruction. (Xiong et al. Nature Methods, 2021)

In vitro Embryo culture

We have established a highly effective embryo culture system for heart live-imaging.

We removed the yolk sac, EPC and amnion of E8.25 embryos, while leave them intact for embryos at E8.0 or earlier. Then embryos were cultured in the petri dish or V-Slot. Morphological, histological and molecular analyses confirm that our culture conditions were able to support apparently normal heart development for more than 24 h with initiating stages from E6.5–E9.0, covering the developmental window from the early cardiac crescent stage (E7.5) to the early four-chambered stage (E10.0).

In toto live imaging

Time-resolved in toto cell lineage tracing.

Time-resolved in toto cell lineage tracing is fundamental for understanding of the intercellular emergent properties for mammalian organogenesis. Having assembling an interdisciplinary group, we developed a customized vertical light-sheet microscope (vLSFM) equipped with an integrated embryo culture module, a heartbeat-gated imaging strategy, and a digital image processing framework for segmentation, tracking, and lineaging. Our integrated light-sheet microscopy-embryo culture system enables single-cell recording of pre-implantation to organogenesis. (Yue et al. Nature Cell Biology, 2020)