A5044985085- CRISPR and Genetic Engineering
- Virus-based gene therapy research
- RNA and protein synthesis mechanisms
- CAR-T cell therapy research
- Hemoglobinopathies and Related Disorders
- Prenatal Screening and Diagnostics
- Advanced biosensing and bioanalysis techniques
- Mosquito-borne diseases and control
- RNA regulation and disease
- Cystic Fibrosis Research Advances
- Pluripotent Stem Cells Research
- Energy Harvesting in Wireless Networks
- Viral Infections and Immunology Research
- Innovation and Socioeconomic Development
- Cytomegalovirus and herpesvirus research
- RNA Interference and Gene Delivery
- Evolution and Genetic Dynamics
- RNA modifications and cancer
- Genomics and Phylogenetic Studies
- Viral Infectious Diseases and Gene Expression in Insects
- Single-cell and spatial transcriptomics
- Immune Cell Function and Interaction
- Bacteriophages and microbial interactions
- Neonatal Respiratory Health Research
- Lysosomal Storage Disorders Research
University College Cork
2012-2025
Rice University
2015-2024
APC Microbiome Institute
2019-2024
National University of Ireland
2024
Georgia Institute of Technology
2014-2018
Emory University
2014-2018
Jackson Laboratory
2017
Ewha Womans University
2017
The Wallace H. Coulter Department of Biomedical Engineering
2016
Texas Medical Center
2016
Precise genome editing using engineered nucleases can significantly facilitate biological studies and disease treatment. In particular, clustered regularly interspaced short palindromic repeats (CRISPR) with CRISPR-associated (Cas) proteins are a potentially powerful tool for modifying by targeted cleavage of DNA sequences complementary to designed guide strand RNAs. Although CRISPR/Cas systems have on-target rates close the transfection rates, they may also relatively high off-target at...
RNA-guided nucleases (RGNs) based on the type II CRISPR-Cas9 system of Streptococcus pyogenes (Sp) have been widely used for genome editing in experimental models. However, nontrivial level off-target activity reported several human cells may hamper clinical translation. RGN specificity depends both guide RNA (gRNA) and protospacer adjacent motif (PAM) recognized by Cas9 protein. We hypothesized that more stringent PAM requirements reduce occurrence mutagenesis. To test this postulation, we...
The clustered regularly-interspaced short palindromic repeats (CRISPR)—CRISPR-associated (Cas) system from Streptococcus pyogenes (Spy) has been successfully adapted for RNA-guided genome editing in a wide range of organisms. However, numerous reports have indicated that Spy CRISPR-Cas9 systems may significant off-target cleavage genomic DNA sequences differing the intended on-target site. Here, we report performance Neisseria meningitidis (Nme) requires longer protospacer-adjacent motif...
Adeno-associated viral (AAV) vectors are a leading candidate for the delivery of CRISPR-Cas9 therapeutic genome editing in vivo. However, AAV-based involves persistent expression Cas9 nuclease, bacterial protein. Recent studies indicate high prevalence neutralizing antibodies and T cells specific to commonly used orthologs from Streptococcus pyogenes (SpCas9) Staphylococcus aureus (SaCas9) humans. We tested mouse model whether pre-existing immunity SaCas9 would pose barrier liver with AAV...
Abstract Gene correction in human long-term hematopoietic stem cells (LT-HSCs) could be an effective therapy for monogenic diseases of the blood and immune system. Here we describe approach X-linked sSevere cCombined iImmunodeficiency (SCID-X1) using targeted integration a cDNA into endogenous start codon to functionally correct disease-causing mutations throughout gene. Using CRISPR-Cas9/AAV6 based strategy, achieve up 20% frequencies LT-HSCs. As measures lack toxicity observe no evidence...
Preclinical efficacy and safety data in mice provide support for ex vivo β-globin gene correction to treat patients with sickle cell disease.
Abstract Rewiring of host cytokine networks is a key feature inflammatory bowel diseases (IBD) such as Crohn’s disease (CD). Th1-type cytokines—IFN-γ and TNF-α—occupy critical nodes within these both are associated with disruption gut epithelial barrier function. This may be due to their ability synergistically trigger the death intestinal cells (IECs) via largely unknown mechanisms. In this study, through unbiased kinome RNAi drug repurposing screens we identified JAK1/2 kinases principal...
Abstract Sickle cell disease (SCD) is a monogenic disorder that affects millions worldwide. Allogeneic hematopoietic stem transplantation the only available cure. Here, we demonstrate use of CRISPR/Cas9 and short single-stranded oligonucleotide template to correct sickle mutation in β-globin gene progenitor cells (HSPCs) from peripheral blood or bone marrow patients with SCD, 24.5 ± 7.6% efficiency without selection. Erythrocytes derived gene-edited showed marked reduction cells, level...
The CRISPR/Cas9 system has been applied in diverse eukaryotic organisms for targeted mutagenesis. However, gene editing is inefficient and requires the simultaneous delivery of a DNA template homology-directed repair (HDR). Here, we used to generate double-strand breaks deliver an RNA HDR rice (Oryza sativa). We chimeric single-guide (cgRNA) molecules carrying both sequences target site specificity (to breaks) direct HDR), flanked by regions homology target. Gene was more efficient...
Editing the β-globin locus in hematopoietic stem cells is an alternative therapeutic approach for gene therapy of β-thalassemia and sickle cell disease. Using CRISPR/Cas9 system, we genetically modified human progenitor (HSPCs) to mimic large rearrangements associated with hereditary persistence fetal hemoglobin (HPFH), a condition that mitigates clinical phenotype patients β-hemoglobinopathies. We optimized compared efficiency plasmid-, lentiviral vector (LV)-, RNA-, ribonucleoprotein...
Adeno-associated viral (AAV) vectors packaging the CRISPR-Cas9 system (AAV-CRISPR) can efficiently modify disease-relevant genes in somatic tissues with high efficiency. AAV are a preferred delivery vehicle for tissue-directed gene therapy because of their ability to achieve sustained expression from largely non-integrating episomal genomes. However, genome editizng applications, permanent non-human proteins such as bacterially derived Cas9 nuclease is undesirable. Methods needed efficient...
Abstract Lysosomal enzyme deficiencies comprise a large group of genetic disorders that generally lack effective treatments. A potential treatment approach is to engineer the patient’s own hematopoietic system express high levels deficient enzyme, thereby correcting biochemical defect and halting disease progression. Here, we present an efficient ex vivo genome editing using CRISPR-Cas9 targets lysosomal iduronidase CCR5 safe harbor locus in human CD34+ stem progenitor cells. The modified...
CRISPR-Cas9–edited hematopoietic stem cells produce long-term engraftment and fetal hemoglobin reactivation in nonhuman primates.
Gene editing of FOXP3 ensures regulated expression and restored function in T cells, supporting clinical applicability.
Abstract Germline manipulation using CRISPR/Cas9 genome editing has dramatically accelerated the generation of new mouse models. Nonetheless, many metabolic disease models still depend upon laborious germline targeting, and are further complicated by need to avoid developmental phenotypes. We sought address these experimental limitations generating somatic mutations in adult liver CRISPR/Cas9, as a strategy model disorders. As proof-of-principle, we targeted low-density lipoprotein receptor...
The use of zinc-finger nucleases (ZFNs) to permanently and precisely modify the human genome offers a potential alternative cDNA-based gene therapy. ΔF508 mutation in cystic fibrosis transmembrane conductance regulator (CFTR) is observed ∼70% patients with (CF) candidate for ZFN-mediated repair. Here, we report modular design synthesis pair ZFNs that can create double-stranded break (DSB) 203 bp upstream lesion, resulting nonhomologous end-joining (NHEJ) frequency 7.8%. In spite this...