Advanced tools: Preclinical mouse models, Single cell technics and HuluFISH

Advanced tools: mouse models, single cell technics and HuluFISH

Preclinical mouse models of glioma

A huge amount of data are being collected from patient samples, however, most of these information will be only considered as descriptive results if they are not tested in experimental setting using model system. To provide better mouse models of human brain cancer for this purpose, we are applying several sets of genetic tools including Cre/LoxP, somatic gene transfer and CRISPR/CAS9 to generate mouse models that mimic both the phenotypic and genetic heterogeneity from human patients. The goal is to provide mouse models can be used to study tumor initiation, tumor genetic evolution and preclinical target validation in primary tumor models.

Single cell technics

The human body is made of individual cells, the ability of performing research on single cell resolution is the next frontier in biology.  Recent years, we investigated a lot of effort on developing and establishing tools for performing quantitative single cell tracing, single cell genetic manipulations and single cell RNA-sequencing. This set of tools will enable us to perform functional studies on single cell level in vivo.

HuluFISH

Single molecule fluorescence in situ hybridization (smFISH) is the most direct way of visualization and quantification of gene expressions without loosing spatial information. However, the research community is still lacking an efficient and highly flexible smFISH method, in particular for multiplexing RNA detections. Thus this technic is still not widely applied in the research community. We developed a novel approach to generate smFISH probes using combinatorial color barcoding strategy (termed HuluFISH). HuluFISH utilizes a highly efficient enzymatic ligation approach to generate smFISH probes conjugated with fully customized fluorophore combinations. This enables robust multiplexing detection of different RNAs in single hybridization without any further intermediate steps.  The multiplex capacity of HuluFISH follows an exponential increase with the increase of the number of fluorophore types. We are further advancing the capacity of HuluFISH and using it to investigate molecular regulations of brain development.

HuluFISH
© Dr. Haikun Liu

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