A5070001273- CRISPR and Genetic Engineering
- HIV Research and Treatment
- Polyomavirus and related diseases
- Full-Duplex Wireless Communications
- Cytomegalovirus and herpesvirus research
- Virus-based gene therapy research
- RNA Interference and Gene Delivery
- Mosquito-borne diseases and control
- Plant Virus Research Studies
- Antenna Design and Analysis
- MicroRNA in disease regulation
- Pluripotent Stem Cells Research
- HIV/AIDS Research and Interventions
- Viral gastroenteritis research and epidemiology
- Immune Cell Function and Interaction
- HIV-related health complications and treatments
- Cancer-related Molecular Pathways
- Neurogenesis and neuroplasticity mechanisms
- RNA modifications and cancer
- DNA Repair Mechanisms
- HIV/AIDS drug development and treatment
- Cannabis and Cannabinoid Research
- Innovation and Socioeconomic Development
- NF-κB Signaling Pathways
- HIV/AIDS Impact and Responses
Temple University
2016-2025
Jagiellonian University
2024
AIDS remains incurable due to the permanent integration of HIV-1 into host genome, imparting risk viral reactivation even after antiretroviral therapy. New strategies are needed ablate genome from latently infected cells, because current methods too inefficient and prone adverse off-target effects. To eliminate integrated we used Cas9/guide RNA (gRNA) system, in single multiplex configurations. We identified highly specific targets within LTR U3 region that were efficiently edited by...
Abstract We employed an RNA-guided CRISPR/Cas9 DNA editing system to precisely remove the entire HIV-1 genome spanning between 5′ and 3′ LTRs of integrated proviral copies from latently infected human CD4+ T-cells. Comprehensive assessment whole-genome sequencing eradicated cells ruled out any off-target effects by our technology that might compromise integrity host further showed no effect on several cell health indices including viability, cycle apoptosis. Persistent co-expression Cas9...
Abstract Elimination of HIV-1 requires clearance and removal integrated proviral DNA from infected cells tissues. Here, sequential long-acting slow-effective release antiviral therapy (LASER ART) CRISPR-Cas9 demonstrate viral in latent infectious reservoirs humanized mice. subgenomic fragments, spanning the long terminal repeats Gag gene, are excised vivo, resulting elimination DNA; virus is not detected blood, lymphoid tissue, bone marrow brain by nested digital-droplet PCR as well RNAscope...
Treatment of HIV-1 ADA -infected CD34+ NSG-humanized mice with long-acting ester prodrugs cabotegravir, lamivudine, and abacavir in combination native rilpivirine was followed by dual CRISPR-Cas9 C-C chemokine receptor type five (CCR5) proviral DNA gene editing. This led to sequential viral suppression, restoration absolute human CD4 + T cell numbers, then elimination replication-competent virus 58% infected mice. Dual CRISPR therapies enabled the excision integrated cells contained within...
Abstract Current antiretroviral therapy does not eliminate the integrated and transcriptionally silent HIV-1 provirus in latently infected cells. Recently, a “shock kill” strategy has been extensively explored to eradicate latent reservoirs for permanent cure of AIDS. The therapeutic efficacy currently used agents remains disappointing because low efficiency, non-specificity cellular toxicity. Here we present novel catalytically-deficient Cas9-synergistic activation mediator (dCas9-SAM)...
Abstract Elimination of HIV DNA from infected individuals remains a challenge in medicine. Here, we demonstrate that intravenous inoculation SIV-infected macaques, well-accepted non-human primate model infection, with adeno-associated virus 9 (AAV9)-CRISPR/Cas9 gene editing construct designed for eliminating proviral SIV DNA, leads to broad distribution molecules and precise cleavage removal fragments the integrated genome blood cells tissues known be viral reservoirs including lymph nodes,...
Progressive multifocal leukoencephalopathy (PML) is a fatal demyelinating disease of the central nervous system (CNS) caused by reactivation human polyomavirus JCV gene expression and its replication in oligodendrocytes, myelin producing cells brain. Once rare seen patients with lymphotproliferative myeloproliferative disorders, PML has been more frequently HIV-1 positive/AIDS as well undergoing immunomodulatory therapy due for autoimmune disorders including multiple sclerosis, rheumatoid...
Abstract In this study, we demonstrate the safety and utility of CRISPR-Cas9 gene editing technology for in vivo proviral DNA ART-treated, virally controlled simian immunodeficiency virus (SIV) infected rhesus macaques, an established model HIV infection. EBT-001 is AAV9-based vector delivering SaCas9 dual guide RNAs designed to target multiple regions SIV genome: viral LTRs, Gag gene. The results presented here that a single IV inoculation at each 3 dose levels (1.4 × 10 12 , 1.4 13 14...
The CRISPR/Cas9 gene editing method is comprised of the guide RNA (gRNA) to target a specific DNA sequence for cleavage and Cas9 endonuclease introducing breaks in double-stranded identified by gRNA. Co-expression both multiplex HIV-1-specific gRNAs cells results modification and/or excision segment viral DNA, leading replication-defective virus. In this study, we have personalized activity placing encoding under control minimal promoter HIV-1 that activated Tat protein. We demonstrate...
Airway basal cells proliferate and regenerate airway epithelium after injury. The first step during epithelial repair is cell proliferation to close the wound. Previously, we demonstrated that homeobox (HOX) A1 expression reduced in stem isolated from chronic obstructive pulmonary disease. HOXA1 a developmental gene plays role hematopoietic differentiation, but its contribution migration not known. In this study, generated knockout bronchial line using CRISPR/CAS9 technology followed by...
Cardiovascular disease is a leading cause of co-morbidity in HIV-1 positive patients, even those whom plasma virus levels are well-controlled. The pathogenic mechanism HIV-1-associated cardiomyopathy unknown, but has been presumed to be mediated indirectly, owing the absence productive replication cardiomyocytes. We sought investigate effect auxiliary protein, Nef, which suspected extracellular release by infected CD4+ T cells on protein quality control and autophagy After detection Nef...
Post-translational glycosylation of the HIV-1 envelope protein involving precursor glycan trimming by mannosyl oligosaccharide glucosidase (MOGS) is critically important for morphogenesis virions and viral entry. Strategic editing MOGS gene in T lymphocytes myeloid origin cells harboring latent proviral DNA results production non-infectious particles upon treatment with latency reversal agents. Controlled activation CRISPR-MOGS rebound mitigates infectious that exhibit poor ability virus to...
Abstract An indispensable role for oligodendrocytes in the protection of axon function and promotion neuronal survival is strongly supported by finding progressive neuron/axon degeneration human neurological diseases that affect oligodendrocytes. Imaging pathological studies CNS have shown presence neuroaxonal injury multifocal leukoencephalopathy (PML), a demyelinating disease CNS, resulting from destruction upon productive replication pathogenic neurotropic polyomavirus JC. Here, we...
ABSTRACT Productive infection of oligodendrocytes, which are responsible for the formation myelin sheath in central nervous system, with human neurotropic virus JC (JCV) causes fatal demyelinating disease progressive multifocal leukoencephalopathy (PML). In addition to encoding T antigen and capsid proteins, produced at early late phases cycle, respectively, JCV encodes a small regulatory protein named agnoprotein that is important successful completion life cycle. Here we used bipotential...
Abstract Reactivation of the human polyomavirus JC (JCV) in CNS results a fatal demyelinating disease, progressive multifocal leukoencephalopathy (PML). The lytic destruction oligodendrocytes, which occurs at terminal stage viral infection cycle, is considered critical factor development demyelination and pathogenesis PML. However, knowledge limited about interaction JCV with oligodendrocytes its impact on denudation axons early reactivation prior to infected cells. We have developed an...