In JoVE (1)
Other Publications (1)
Articles by Nitin Chakravarti in JoVE
Generation of Knock-out Primary and Expanded Human NK Cells Using Cas9 Ribonucleoproteins Meisam Naeimi Kararoudi1, Hamid Dolatshad2, Prashant Trikha1, Syed-Rehan A. Hussain3, Ezgi Elmas1, Jennifer A. Foltz1, Jena E. Moseman1, Aarohi Thakkar1, Robin J. Nakkula1, Margaret Lamb1, Nitin Chakravarti1, K. John McLaughlin3, Dean A. Lee1 1 Here, we present a protocol to genetically modify primary or expanded human natural killer (NK) cells using Cas9 Ribonucleoproteins (Cas9/RNPs). By using this protocol, we generated human NK cells deficient for transforming growth factor–b receptor 2 (TGFBR2) and hypoxanthine phosphoribosyltransferase 1 (HPRT1).
Other articles by Nitin Chakravarti on PubMed
Diminished MicroRNA-29b Level is Associated with BRD4-mediated Activation of Oncogenes in Cutaneous T-cell Lymphoma Blood. Feb, 2018 | Pubmed ID: 29180399 MicroRNA (miRNA) dysregulation is a hallmark of cutaneous T-cell lymphoma (CTCL), an often-fatal malignancy of skin-homing CD4 T cells for which there are few effective therapies. The role of microRNAs (miRs) in controlling epigenetic modifier-dependent transcriptional regulation in CTCL is unknown. In this study, we characterize a novel miR dysregulation that contributes to overexpression of the epigenetic reader bromodomain-containing protein 4 (BRD4). We used patient CD4 T cells to show diminished levels of miR-29b compared with healthy donor cells. Patient cells and miR-29b mouse cells revealed an inverse relationship between miR-29b and BRD4, the latter of which is overexpressed in these cells. Chromatin immunoprecipitation and sequencing analysis revealed increased genome-wide BRD4 occupancy at promoter and enhancer regions in CD4 T cells from CTCL patients. The cumulative result of BRD4 binding was increased expression of tumor-associated genes such as and , as well as the interleukin-15 (IL-15) receptor complex, the latter enhancing IL-15 autocrine signaling. Furthermore, we confirm the in vivo relevance of this pathway in our IL-15 transgenic mouse model of CTCL by showing that interference with BRD4-mediated pathogenesis, either by restoring miR-29b levels via bortezomib treatment or by directly inhibiting BRD4 binding via JQ1 treatment, prevents progression of CTCL. We describe a novel oncogenic pathway featuring IL-15, miR-29b, and BRD4 in CTCL and suggest targeting of these components as a potentially effective therapy for CTCL patients.