A5082099200- CRISPR and Genetic Engineering
- Virus-based gene therapy research
- RNA Interference and Gene Delivery
- interferon and immune responses
- Animal Genetics and Reproduction
- HIV Research and Treatment
- Advanced biosensing and bioanalysis techniques
- Viral Infections and Immunology Research
- Lymphoma Diagnosis and Treatment
- Retinal Development and Disorders
- Cytomegalovirus and herpesvirus research
- MicroRNA in disease regulation
- Chromosomal and Genetic Variations
- Herpesvirus Infections and Treatments
- RNA regulation and disease
- CAR-T cell therapy research
- Circular RNAs in diseases
- RNA modifications and cancer
- Immune Response and Inflammation
- Pluripotent Stem Cells Research
- Immune Cell Function and Interaction
- NF-κB Signaling Pathways
- Viral Infections and Vectors
- Viral Infectious Diseases and Gene Expression in Insects
- Retinal Diseases and Treatments
Aarhus University
2015-2024
Institute for Biomedicine
2010-2020
Innate Pharma (France)
2015
Aarhus University Hospital
2014
Rigshospitalet
2014
International AIDS Vaccine Initiative
2013
Scripps Research Institute
2013
Pediatrics and Genetics
2003-2007
Stanford University
2002-2007
Significance HIV-1 is a lentivirus and replicates through replication cycle involving several DNA forms including ssDNA. Here we report that synthetic oligos corresponding to of the as well viral are detected by immunological sensor IFN-inducible protein 16 (IFI16) stimulate innate immune responses pathway dependent on stimulator IFN genes (STING). Moreover, show elevated in cells with decreased expression IFI16 or STING. We suggest for macrophages stimulating responses, which contribute...
Abstract Innate immune activation by macrophages is an essential part of host defence against infection. Cytosolic recognition microbial DNA in leads to induction interferons and cytokines through cyclic GMP-AMP synthase (cGAS) stimulator interferon genes (STING). Other factors, including interferon-gamma inducible factor 16 (IFI16), have been proposed contribute DNA. However, their relation the cGAS-STING pathway not clear. Here, we show that IFI16 functions on two distinct levels....
Atherosclerosis can be achieved in animals by germline genetic engineering, leading to hypercholesterolemia, but such models are constrained few species and strains, they difficult combine with other powerful techniques involving manipulation or variation.To develop a method for induction of atherosclerosis without engineering.Recombinant adeno-associated viral vectors were engineered encode gain-of-function proprotein convertase subtilisin/kexin type 9 mutants, mice given single intravenous...
Abstract Stimulator of interferon genes (STING) is known be involved in control DNA viruses but has an unexplored role RNA viruses. During infection with STING activated downstream cGAMP synthase (cGAS) to induce type I interferon. Here we identify a STING-dependent, cGAS-independent pathway important for full production and antiviral enveloped viruses, including influenza A virus (IAV). Further, IAV interacts through its conserved hemagglutinin fusion peptide (FP). Interestingly, FP...
Herpes simplex encephalitis (HSE) in children has previously been linked to defects type I interferon (IFN) production downstream of Toll-like receptor 3. Here, we describe a novel genetic etiology HSE by identifying heterozygous loss-of-function mutation the IFN regulatory factor 3 (IRF3) gene, leading autosomal dominant (AD) IRF3 deficiency haploinsufficiency, an adolescent female patient with HSE. is activated most pattern recognition receptors recognizing viral infections and plays...
A transgenic pig model of familial hypercholesterolemia can be used for translational atherosclerosis research.
Hepatitis B virus (HBV) is a major human pathogen, and about one third of the global population will be exposed to in their lifetime. HBV infects hepatocytes, where it replicates its DNA infection can lead acute chronic hepatitis with high risk liver cirrhosis hepatocellular carcinoma. Despite this, there limited understanding how establishes infections. In recent years has emerged that foreign potently stimulates innate immune response, particularly type 1 interferon (IFN) production; this...
Varicella zoster virus (VZV) typically causes chickenpox upon primary infection. In rare cases, VZV can give rise to life-threatening disease in otherwise healthy people, but the immunological basis for this remains unexplained. We report 4 cases of acute severe infection affecting central nervous system or lungs unrelated, children who are heterozygous missense mutations POLR3A (one patient), POLR3C both (two patients). and encode subunits RNA polymerase III. Leukocytes from all patients...
Future therapeutic use of engineered site-directed nucleases, like zinc-finger nucleases (ZFNs) and transcription activator-like effector (TALENs), relies on safe effective means delivering to cells. In this study, we adapt lentiviral vectors as carriers designer nuclease proteins, providing efficient targeted gene disruption in vector-treated cell lines primary By co-packaging pairs ZFN proteins with donor RNA ‘all-in-one’ particles, co-deliver the template for homology-directed repair...
Type I interferons (IFN-I) play a critical role in human antiviral immunity, as demonstrated by the exceptionally rare deleterious variants of IFNAR1 or IFNAR2. We investigated five children from Greenland, Canada, and Alaska presenting with viral diseases, including life-threatening COVID-19 influenza, addition to meningoencephalitis and/or hemophagocytic lymphohistiocytosis following live-attenuated vaccination. The affected individuals bore same homozygous IFNAR2 c.157T>C,...
Prime editing is a new CRISPR-based, genome-editing technology that relies on the prime editor (PE), fusion protein of Cas9-nickase and M-MLV reverse transcriptase (RT), guide RNA (pegRNA) serves both to target PE desired genomic locus carry edit be introduced. Here, we make advancements RT moiety improve efficiencies truncations mitigate issues with adeno-associated virus (AAV) viral vector size limitations, which currently do not support efficient delivery large components. These efforts...
Implementation of therapeutic in vivo gene editing using CRISPR/Cas relies on potent delivery tools. Administration ribonucleoprotein (RNP) complexes consisting Cas protein and single guide RNA (sgRNA) offers short-lived activity safety advantages over conventional viral non-viral approaches. By engineering lentivirus-derived nanoparticles (LVNPs) to facilitate RNP delivery, we demonstrate effective administration SpCas9 as well SpCas9-derived base prime editors (BE/PE) leading recipient...
The N-terminal domain of the Sleeping Beauty (SB) transposase mediates transposon DNA binding, subunit multimerization, and nuclear translocation in vertebrate cells. For this report, we studied relative contributions 95 different residues within multifunctional by large-scale mutational analysis. We found that each four amino acids (leucine 25, arginine 36, isoleucine 42, glycine 59) contributes to binding context 123 SB transposase, as indicated electrophoretic mobility shift analysis,...
Transposon-based vectors represent promising new tools for chromosomal transgene insertion and establishment of persistent gene expression in vivo. Here, we report the development helper-independent transposon-transposase (HITT) vectors, which contain on single plasmids (i) a Sleeping Beauty (SB) transposon containing (ii) SB transposase cassette. To obtain an optimal level from HITT determined relative strength panel different promoters mouse liver used these to drive injected carrying...
Gene delivery by human immunodeficiency virus type 1 (HIV-1)-based lentiviral vectors (LVs) is efficient, but genomic integration of the viral DNA strongly biased toward transcriptionally active loci resulting in an increased risk insertional mutagenesis gene therapy protocols. Nonviral Sleeping Beauty (SB) transposon have a significantly safer insertion profile, efficient into relevant cell/tissue types limitation. In attempt to combine favorable features two vector systems we established...
It has been previously shown that integrase-defective HIV-1-based gene vectors can serve, with moderate efficiency, as substrate for DNA transposition by a transiently expressed Sleeping Beauty (SB) transposase. Here, we describe the enhanced transfer properties of HIV-1/SB hybrid vector allows efficient transposition, facilitated hyperactive SB100X transposase, from intermediates in primary human cells. Potent transposase-dependent integration genetic cargo carried (up to 160-fold above...
Virus-based gene therapy by CRISPR/Cas9-mediated genome editing and knockout may provide a new option for treatment of inherited acquired ocular diseases the retina. In support this notion, we show that Streptococcus pyogenes (Sp) Cas9, delivered lentiviral vectors (LVs), can be used in vivo to selectively ablate vascular endothelial growth factor A (Vegfa) mice. By generating LVs encoding SpCas9 targeted Vegfa, parallel fluorescent eGFP marker protein, demonstrate robust Vegfa leads...
Tumor necrosis factor-alpha (TNF-alpha) is upregulated in psoriatic skin and represents a prominent target psoriasis treatment. The level of TNF-alpha-encoding mRNA, however, not increased skin, it remains unclear whether intervention strategies based on RNA interference (RNAi) are therapeutically relevant. To test this hypothesis the present study describes first vitro functional screening panel short hairpin RNAs (shRNAs) targeting human TNF-alpha mRNA and, next, transfer most potent shRNA...
MicroRNAs (miRNAs) are key regulators of gene expression and modulators diverse biological pathways. Analyses miRNA function as well therapeutic managing miRNAs rely on cellular administration inhibitors which may be achieved by the use viral vehicles. This study explores miRNA-suppressive capacity expressed intracellularly from lentivirus-derived vectors. Superior activity two decoy-type inhibitors, a “Bulged Sponge” with eight recognition sites hairpin-shaped “Tough Decoy” containing...