A5064427220- CRISPR and Genetic Engineering
- DNA Repair Mechanisms
- RNA Research and Splicing
- RNA and protein synthesis mechanisms
- Genomics and Chromatin Dynamics
- RNA modifications and cancer
- Telomeres, Telomerase, and Senescence
- Animal Virus Infections Studies
- Herpesvirus Infections and Treatments
- T-cell and B-cell Immunology
- Genetic Neurodegenerative Diseases
- Immune Cell Function and Interaction
- Mitochondrial Function and Pathology
- Virus-based gene therapy research
- Cancer therapeutics and mechanisms
- Cellular transport and secretion
- Immune Response and Inflammation
- Cancer-related molecular mechanisms research
- Polyomavirus and related diseases
- T-cell and Retrovirus Studies
- Animal Disease Management and Epidemiology
- Ferroptosis and cancer prognosis
- Plant Virus Research Studies
- Advanced biosensing and bioanalysis techniques
- Genomics, phytochemicals, and oxidative stress
University of Minnesota
1994-2024
University of Minnesota Medical Center
1998-2023
The use of programmable meganucleases is transforming genome editing and functional genomics. CRISPR/Cas9 was developed such that targeted genomic lesions could be introduced in vivo with unprecedented ease. In the presence homology donors, these facilitate high-efficiency precise (PGE) via homology-directed repair (HDR) pathways. However, identity hierarchy HDR (sub)pathways leading to formation PGE products remain elusive. Here, we established a green blue fluorescent protein conversion...
The DNA-dependent protein kinase (DNA-PK) complex is a serine/threonine comprised of 469-kDa catalytic subunit (DNA-PKcs) and the DNA binding regulatory heterodimeric (Ku70/Ku86) Ku. DNA-PK functions in nonhomologous end-joining pathway for repair double-stranded breaks (DSBs) introduced by either exogenous damage or endogenous processes, such as lymphoid V(D)J recombination. Not surprisingly, mutations Ku70, Ku86, DNA-PKcs result animals that are sensitive to agents cause DSBs also immune...
Gene targeting in human somatic cells is of importance because it can be used to either delineate the loss-of-function phenotype a gene or correct mutated back wild-type. Both these outcomes require form DNA double-strand break (DSB) repair known as homologous recombination (HR). The mechanism HR leading targeting, however, not well understood cells. Here, we demonstrate that two-end, ends-out intermediate valid for targeting. Furthermore, resolution step this occurs via classic DSB model...
Telomeres shorten with each cell division and can ultimately become substrates for nonhomologous end-joining repair, leading to large-scale genomic rearrangements of the kind frequently observed in human cancers. We have characterized more than 1400 telomere fusion events at single-molecule level, using a combination high-throughput sequence analysis together experimentally induced telomeric double-stranded DNA breaks. show that single chromosomal dysfunctional fuse diverse nontelomeric...
A new subgroup of avian leukosis virus (ALV), designated J, was identified recently. Viruses this do not cross-interfere with viruses the A, B, C, D, and E subgroups, are neutralized by antisera raised against other have a broader host range than to subgroups. Sequence comparisons reveal that while J envelope gene includes some regions related those found in env genes majority is composed sequences either more similar member (E51) ancient endogenous (EAV) family proviruses or appear unique...
Fusion of critically short or damaged telomeres is associated with the genomic rearrangements that support malignant transformation. We have demonstrated fundamental contribution DNA ligase 4-dependent classical non-homologous end-joining to long-range inter-chromosomal telomere fusions. In contrast, localized recombinations initiated by sister chromatid fusion are predominantly mediated alternative activity may employ either 3 1. this study, we sought discriminate relative involvement these...
The Artemis nuclease and tyrosyl-DNA phosphodiesterase (TDP1) are each capable of resolving protruding 3'-phosphoglycolate (PG) termini DNA double-strand breaks (DSBs). Consequently, both a knockout knockout/knockdown TDP1 rendered cells sensitive to the radiomimetic agent neocarzinostatin (NCS), which induces 3'-PG-terminated DSBs. Unexpectedly, however, knockdown or in Artemis-null did not confer any greater sensitivity than either deficiency alone, indicating strict epistasis between...
Protein phosphatase I (PP1) is an essential eukaryotic serine/threonine required for many cellular processes, including cell division, signaling, and metabolism. In mammalian cells there are three major isoforms of the PP1 catalytic subunit (PP1alpha, PP1beta, PP1gamma) that over 90% identical. Despite this high degree identity, subunits show distinct localization patterns in interphase cells; PP1alpha primarily nuclear largely excluded from nucleoli, whereas PP1gamma to a lesser extent...
A new subgroup of avian leukosis virus (ALV) that includes a unique env gene, designated J, was identified recently in England. Sequence analysis prototype English isolate HPRS-103 revealed several other genetic characteristics this strain and provided information it arose by recombination between exogenous endogenous sequences. In the past years, ALV J type viruses (ALV-J) have been isolated from broiler breeder flocks United States. We were interested determining relationship U.S. isolates...
Here, we report that the LynB splice variant of Src-family kinase Lyn exerts a dominant immunosuppressive function in vivo, whereas LynA isoform is uniquely required to restrain autoimmunity female mice. We used CRISPR-Cas9 gene editing constrain lyn splicing and expression, generating single-isoform knockout (LynA KO ) or Autoimmune disease total mice characterized by production antinuclear antibodies, glomerulonephritis, impaired B cell development, overabundance activated cells...
Virtually all pre-mRNA introns begin with the sequence / GU and end AG/ (where indicates a border between an exon Intron). We have previously shown that G residues at first last positions of yeast act In Intron Interact during second step splicing. this work, we ask If other highly conserved nucleotides also take part - G/ interaction. Of special interest is penultimate nucleotlde (AGl), which Is important for splicing proximity to intron nucleotides. Therefore, tested interactions...
ABSTRACT We recently reported the identification of sequences in chicken genome that show over 95% identity to novel envelope gene subgroup J avian leukosis virus (S. J. Benson, B. L. Ruis, A. M. Fadly, and K. F. Conklin, Virol. 72:10157–10164, 1998). Based on fact endogenous J-related env genes were associated with long terminal repeats (LTRs), we concluded these LTR- defined a new family viruses designated ev/J family. In this report, have further characterized content expression...
Abstract The unique roles of the Src-family kinases LynA and LynB in immune activating inhibitory signaling have eluded definition. Here we report that LynB, shorter splice product lyn , carries dominant immunosuppressive function. We used CRISPR/Cas9 gene editing to constrain splicing expression a single product: KO or mice. While activities both isoforms regulate homeostatic Lyn expression, only protects against autoimmune disease. monocytes dendritic cells are TLR4-hyper-responsive, TLR4...
Abstract Ten-Eleven Translocation (TET1-3) dioxygenases oxidize 5-methylcytosine (5mC) in DNA to generate 5-hydroxymethylcytosine (5hmC), 5-formylcytosine (5fC), and 5-carboxylcytosine (5caC), initiating demethylation. The three proteins share significant sequence homology catalyze the same chemical reaction utilizing alpha-ketoglutarate cofactor non-heme iron methyl group of 5mC. Since their discovery 2009, there have been contradictory reports regarding roles TET cancer. genes...
Prime editing brings immense promise to correct a large number of human pathogenic mutations and enact diverse edit types without introducing widespread undesired events. Delivery prime editors in vivo would enable such edits be introduced clinical setting. The coding sequence for editor, however, is too fit within the size-constrained adeno-associated virus (AAV) genome. Herein, we describe split Staphylococcus aureus editor capable being delivered by dual AAVs. We characterize ability...
SUMMARY Spinocerebellar ataxia type 1 (SCA1) is a dominant trinucleotide repeat neurodegenerative disease characterized by motor dysfunction, cognitive impairment, and premature death. Degeneration of cerebellar Purkinje cells frequent prominent pathological feature SCA1. We previously showed that transport ATXN1 to cell nuclei required for pathology, where mutant alters transcription. To examine the role nuclear localization broadly in SCA1-like pathogenesis, CRISPR-Cas9 was used develop...