A5075520749- CRISPR and Genetic Engineering
- Cancer-related gene regulation
- Epigenetics and DNA Methylation
- RNA modifications and cancer
- Pluripotent Stem Cells Research
- Advanced biosensing and bioanalysis techniques
- Cancer-related molecular mechanisms research
- Acute Myeloid Leukemia Research
- RNA and protein synthesis mechanisms
- RNA Interference and Gene Delivery
- Genetics and Neurodevelopmental Disorders
- Mesenchymal stem cell research
- DNA Repair Mechanisms
- Multiple Myeloma Research and Treatments
- Erythrocyte Function and Pathophysiology
- Protein Degradation and Inhibitors
- Glycosylation and Glycoproteins Research
- Computational Physics and Python Applications
- CAR-T cell therapy research
- Ubiquitin and proteasome pathways
- Immune Cell Function and Interaction
- Immunodeficiency and Autoimmune Disorders
- Genetic Syndromes and Imprinting
- Tissue Engineering and Regenerative Medicine
- Virus-based gene therapy research
Indiana University – Purdue University Indianapolis
2023-2025
Indiana University School of Medicine
2023-2025
University of Indianapolis
2025
Max Delbrück Center
2016-2024
University of Alabama at Birmingham
2014-2021
Oregon Health & Science University
2020-2021
University of Michigan
2020
Kameda Medical Center
2020
Korea Advanced Institute of Science and Technology
2011-2014
The Royal Melbourne Hospital
1997
RBM15, an RNA binding protein, determines cell-fate specification of many tissues including blood. We demonstrate that RBM15 is methylated by protein arginine methyltransferase 1 (PRMT1) at residue R578, leading to its degradation via ubiquitylation E3 ligase (CNOT4). Overexpression PRMT1 in acute megakaryocytic leukemia cell lines blocks megakaryocyte terminal differentiation downregulation level. Restoring level rescues blocked overexpression. At the molecular level, binds pre-messenger...
Significance Efficient clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR associated protein 9 (Cas9)-mediated mutagenesis is necessary for robust genetic screening in primary cells and requires sufficiently high levels of Cas9 reliable single guide RNAs (sgRNAs). We provide a sgRNA design tool that selects high-fidelity sgRNAs transgenic mouse line expresses at all analyzed. Using this system, we achieved an average knockout efficiency 80% B established...
Abstract Overall survival of acute myeloid leukemia (AML) remains limited. Inhibitors the master mitotic kinase PLK1 have emerged as promising therapeutics, demonstrating efficacy in an undefined subset AML patients. However, clinical success inhibitors hindered by a lack predictive biomarkers. The Fanconi anemia (FA) pathway, tumor-suppressive network comprised at least 22 genes, is frequently mutated sporadic AML. Here, we demonstrate that FA pathway disruption sensitizes cells to...
The CRISPR-Cas9 system is used for genome editing in mammalian cells by introducing double-strand breaks (DSBs) which are predominantly repaired via non-homologous end joining (NHEJ) or to lesser extent homology-directed repair (HDR). To enhance HDR improving the introduction of precise genetic modifications, we tested fusion proteins Cas9 nuclease with effectors enforce their localization at DSBs. Using a traffic-light DSB reporter (TLR) quantitative detection and NHEJ events human HEK...
While CRISPR-Cas9 is key for the development of gene therapy, its potential off-target mutations are still a major concern. Here, we establish “spacer-nick” correction approach that combines Cas9 D10A nickase with pair PAM-out sgRNAs at distance 200 to 350 bp. In combination adeno-associated virus (AAV) serotype 6 template delivery, our led efficient HDR in human hematopoietic stem and progenitor cells (HSPCs including long-term HSCs) T cells, minimal NHEJ-mediated on-target mutations. Using...
Severe congenital neutropenia (SCN) is a monogenic disorder. SCN patients are prone to recurrent life-threatening infections. The main causes of autosomal dominant mutations in the ELANE gene that lead block neutrophil differentiation. In this study, we use CRISPR-Cas9 ribonucleoproteins and adeno-associated virus (AAV)6 as donor template delivery system repair ELANEL172P mutation patient-derived hematopoietic stem progenitor cells (HSPCs). We used single guide RNA (sgRNA) specifically...
Mitogen-activated protein kinases (MAPKs) are inactivated by dual-specificity phosphatases (DUSPs), the activities of which tightly regulated during cell differentiation.Using knockdown screening and single-cell transcriptional analysis, we demonstrate that DUSP4 is phosphatase specifically inactivates p38 kinase to promote megakaryocyte (Mk) differentiation.Mechanistically, PRMT1-mediated methylation triggers its ubiquitinylation an E3 ligase HUWE1.Interestingly, mechanistic axis...
Multiple myeloma (MM) is an incurable malignancy characterized by mutated plasma cell clonal expansion in the bone marrow, leading to severe clinical symptoms. Thus, identifying new therapeutic targets for MM crucial. We identified oligosaccharyltransferase (OST) complex as a novel vulnerability cells. Elevated expression of this associated with relapsed, high-risk MM, and poor prognosis. Disrupting OST suppressed growth, induced cell-cycle arrest, apoptosis. Combined inhibition bortezomib...
<p>High-confidence non-synonymous FANCA mutations.</p>
<p>Unbiased profiling of the FANCA interactome across cell cycle identifies PLK1 as a mitosis-specific interacting partner FANCA. <b>A,</b> Schematic illustrating experimental design. HeLa cells were synchronized in S-phase by thymidine block, followed treatment with Ro-3306 to induce G2 arrest. Cells released from into nocodazole or proTAME arrest prometaphase metaphase, respectively. was immunoprecipitated G2, prometaphase, and metaphase protein lysates (two biological...
<div>Abstract<p>Overall survival of acute myeloid leukemia (AML) remains limited. Inhibitors the master mitotic kinase PLK1 have emerged as promising therapeutics, demonstrating efficacy in an undefined subset patients with AML. However, clinical success inhibitors hindered by a lack predictive biomarkers. The Fanconi anemia (FA) pathway, tumor-suppressive network comprised at least 22 genes, is frequently mutated sporadic In this study, we demonstrate that FA pathway disruption...
<p>MIB/MS profiling in AML cell lines reveals PLK1 as sole target of Volasertib at low concentrations.</p>
<p>Low-dose volasertib causes selective toxicity in primary AMLs with predicted pathogenic FA pathway mutations. <b>A,</b> Graph represents viability of patient CD34<sup>+</sup> AML cells treated 10 nmol/L for 5 days. <i>P</i> values were calculated by two‐way ANOVA. <b>B,</b> Panel below bar graph indicates mutations samples identified through WES. <b>C,</b> Gene Ontology and Kyoto Encyclopedia Genes Genomes functional...
<p>FANCA deficiency sensitizes cancer cell lines and primary murine HPCs to PLK1 inhibition. Representative Western blots showing FANCA expression in WT vs. <i>FANCA</i>-KO HeLa (<b>A</b>) Scr.Ctrl FANCA-KD THP-1 (<b>B</b>), HL60 (<b>C</b>), Kasumi-1 (<b>D</b>)<b>,</b> (<b>E</b>) OCI-AML-5 cells. Graphs below each blot represent viability of cells relative control following 3-day treatment with...
<p>Supp Dataset 4</p>
<p>Supp Dataset 2</p>
<p>Supp Dataset 3</p>
<p>FANCD2 co-localizes with PLK1 at centromeres in unperturbed mitotic HeLa cells.</p>
<p>The DDR response of FANCD2 is unaffected after PLK1 inhibition.</p>
<p>Inhibition of PLK1 induces mitotic failure and apoptosis in the FANCA-deficient hTERT-RPE1 cell line. <b>A,</b> Representative Western blot showing FANCA expression control (Scr.Ctrl.) cells vs. FANCA-KD (shFANCA) after 48 hours doxycycline (Dox) treatment. <b>B,</b> Concentration-dependent survival (%) shFANCA Scr.Ctrl treated with volasertib onvansertib for 3 days. Cell viability was measured by CellTiter-Glo assay. graph shows mean ± SEM compared vehicle....
<p>Supp Dataset 1</p>
<p>FANCA-deficiency sensitizes RS4;11 acute lymphoblastic leukemia cell line to PLK1 inhibition.</p>
<p>Supp Dataset 5</p>