A5010459855- Virus-based gene therapy research
- Viral Infectious Diseases and Gene Expression in Insects
- CRISPR and Genetic Engineering
- CAR-T cell therapy research
- RNA Interference and Gene Delivery
- Neuropeptides and Animal Physiology
- Hemoglobinopathies and Related Disorders
- Gastrointestinal motility and disorders
- Diet and metabolism studies
- Pluripotent Stem Cells Research
- Erythrocyte Function and Pathophysiology
- Vector-Borne Animal Diseases
- Genomics and Chromatin Dynamics
- Virology and Viral Diseases
- Animal Disease Management and Epidemiology
- T-cell and Retrovirus Studies
- Reproductive Physiology in Livestock
- T-cell and B-cell Immunology
- Cancer-related gene regulation
- Viral Infections and Outbreaks Research
- Viral gastroenteritis research and epidemiology
- Ruminant Nutrition and Digestive Physiology
- Tryptophan and brain disorders
- Monoclonal and Polyclonal Antibodies Research
- Plant Virus Research Studies
St. Jude Children's Research Hospital
2010-2024
Vector (United States)
2019-2024
Washington University in St. Louis
1997-2005
Instytut Medycyny Wsi im. Witolda Chodźki
1992
Lentiviral vectors are increasingly utilized in cell and gene therapy applications because they efficiently transduce target cells such as hematopoietic stem T cells. Large-scale production of current Good Manufacturing Practices-grade lentiviral is limited the adherent, serum-dependent nature HEK293T used manufacturing process. To optimize large-scale clinical-grade vector production, we developed an improved scheme by adapting to grow suspension using commercially available chemically...
Lentiviral vectors efficiently transduce quiescent stem cells and are being evaluated for gene therapy of blood dis-orders. The risk genotoxicity as a result insertional mutagenesis is an important safety consideration. hy-persensitive site 4 insulator from the chicken beta-globin locus (cHS4) possesses chromatin bar-rier enhancer-blocking functions. A control lentiviral vector encoding green fluorescent protein was compared with in which cHS4 element flanked expression cassette single cell...
We have developed a producer cell line that generates lentiviral vector particles of high titer. The encodes the Wiskott-Aldrich syndrome (WAS) protein. An insulator element has been added to long terminal repeats integrated limit proto-oncogene activation. provides high-level, stable expression WAS protein in transduced murine and human hematopoietic cells. also monoclonal antibody specific for intracellular This used monitor blood bone marrow cells after transfer into lineage negative from...
ABSTRACT The Sindbis virus minimal subgenomic mRNA promoter (spanning positions −19 to +5 relative the start site) is approximately three- sixfold less active than fully −98 +14 region. We identified two elements flanking region which increase its transcription levels comparable These span −40 −20 and +6 act synergistically enhance transcription. Nine different libraries were constructed containing blocks of five randomized nucleotides at various in On passaging these mosquito cells, a small...
Gene therapy for the treatment of Wiskott–Aldrich syndrome (WAS) presents an alternative to current use allogeneic bone marrow transplantation. We describe development a self-inactivating lentiviral vector containing chromatin insulators WAS and compare gammaretroviral (MND), human cellular (EF1α), WASp gene promoter expression patterns in vivo during murine hematopoiesis using green fluorescent protein (GFP) marker. Compared with EF1α promoters, from MND mouse transplant recipients was much...
Cholesterol-rich plasma membrane microdomains are important for entry of many viruses, including retroviruses. Depletion cholesterol with 2-hydroxypropyl-β-cyclodextrin inhibits human T cell leukemia virus type I (HTLV-1) and HTLV-I envelope pseudotyped lentivirus particles. Using a soluble fusion protein the surface immunoglobulin Fc domain, receptor was found to colocalize raft-associated marker cluster in specific microdomains. did not alter binding activity, suggesting requirement...
Lentiviral vectors are useful for transducing primitive hematopoietic cells. We examined four envelope proteins their ability to mediate lentiviral transduction of mobilized human CD34(+) peripheral blood particles encoding green fluorescent protein (GFP) were pseudotyped with the vesicular stomatitis virus glycoprotein (VSV-G), amphotropic (AMPHO) murine leukemia protein, endogenous feline viral or type C protein. Because relative amount genome RNA per ml was similar each pseudotype, we...
Wiskott-Aldrich syndrome (WAS) is a life-threatening immunodeficiency caused by mutations within the WAS gene. Viral gene therapy to restore protein (WASp) expression in hematopoietic cells of patients with has potential improve outcomes relative current standard care, allogeneic bone marrow transplantation. However, development viral vectors that are both safe and effective been problematic. While use transcriptional promoters may increase risk insertional mutagenesis, cellular not achieve...
Sindbis virus (SIN), the type alphavirus, has been studied extensively to identify viral cis-acting sequences and proteins involved in RNA transcription replication. However, very little is known about how these processes are coordinated. For example, synthesis of genomic subgenomic mRNA depends on minus strand. Do activities occur independently different templates, or can replication take place simultaneously same template? We describe appearance a SIN-specific, plus-sense that intermediate...
In xenograft mice and nonhuman primates, BCL11A interference coexpression of a truncated erythropoietin receptor led to sustained hemoglobin F.
Transcriptional enhancers can be in physical proximity of their target genes via chromatin looping. The enhancer at the β-globin locus (locus control region [LCR]) contacts fetal-type (
The human T-cell leukemia virus type 1 (HTLV-1) envelope protein is required for spread. This study further characterizes the role of in HTLV-1 immortalization. Viruses with single amino acid substitutions within SU at residue 75, 81, 95, 101, 105, or 195 a C-terminal cytoplasmic domain truncation (CT), as well an envelope-null (EN) virus, were generated infectious molecular clone, ACH. Transfection 293T cells resulted release similar amounts particles from all mutants determined by p19...
Lentiviral vector gene therapy for hematopoietic disorders caused by single mutation or deficiency has demonstrated success in clinical trials. However, efficient delivery is still challenging, requiring high multiplicity of infection (MOI) to achieve average copy numbers at least 1 per cell. Conditions utilizing MOI result improved transduction rates and adequate number but these benefits can coincide with potential insertional mutagenesis. Attempting optimal transfer reduced MOI, we...
Successful cell and gene therapy clinical trials have resulted in the US Food Drug Administration European Medicines Agency approving their use for treatment of patients with certain types cancers monogenetic diseases. These novel therapies, which rely heavily on lentiviral vectors to deliver therapeutic transgenes patient cells, driven additional investigations, increasing demand both pre-clinical current Good Manufacturing Practices-grade viral vectors. To better support studies by...
Wiskott–Aldrich Syndrome (WAS) is a severe X-linked immunodeficiency disorder characterized by recurrent infection, thrombocytopenia, and autoimmunity resulting from mutations within the WAS gene. The encoded gene product, protein (WASp), scaffold important for activation of actin cytoskeleton downstream multiple hematopoietic receptors. Development safe effective stem cell (HSC) therapy would provide valuable treatment alternative to allogeneic bone marrow transplantation. Of critical...