Project description:Jurkat cell lines were generated to stably express CD46 protein expressing either cytoplasmic tail 1 or 2 (BC1 or BC2). The transcription profile of these unactivated stable lines were compared with unactivated Jurkat cells.
Project description:Vitamin K1 (VK1), an essential nutrient related to coagulation, has been proved to have anticancer properties in various cancer cells. However, little is known about the effects of VK1 on hematologic malignancies. The aim of the study was to evaluate the cytotoxic effects of VK1 on acute T-cell lymphocytic leukemia Jurkat cells (Jurkat T cells). In addition, VK1 induced changes of gene expression in Jurkat T cells were also observed.
Project description:We develop a system for bidirectional epigenetic editing (CRISPRai), in which orthogonal activating (CRISPRa) and repressive (CRISPRi) perturbations are applied simultaneously to multiple loci the same cell. We perform ATAC-seq on CRISPRi perturbed Jurkat T cells to investigate chromatin accessibility changes upon perturbation of individual regulatory elements.
Project description:Tissue-specific gene expression requires coordinated control of gene-proximal and distal cis-regulatory elements (CREs), yet functional analysis of putative gene-distal CREs such as enhancers remains challenging. Here we describe enhanced CRISPR/dCas9-based epigenetic editing systems, enCRISPRa and enCRISPRi, for multiplexed analysis of enhancer function in situ and in vivo. Using dual effector proteins capable of re-writing enhancer-associated chromatin modifications, we show that enCRISPRa and enCRISPRi modulate gene transcription by remodeling local epigenetic landscapes at sgRNA-targeted enhancers and associated genes. Comparing with existing methods, our systems display more robust and specific perturbations of gene transcription with minimal off-targets. Allele-specific targeting of enCRISPRa to oncogenic TAL1 super-enhancer modulates TAL1 expression and cancer progression in xenotransplants. Furthermore, multiplexed perturbations of lineage-specific enhancers in an enCRISPRi knock-in mouse establish in vivo evidence for lineage-restricted requirement of developmentally regulated enhancers during hematopoietic lineage specification. Hence, enhanced CRSIPR epigenetic editing provides opportunities for interrogating enhancer function in development and disease.