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Article: Combining CRISPR-Cas-mediated terminal resolution with a novel genetic workflow to achieve high-diversity adenoviral libraries.

Fischer, Julian / Fedotova, Ariana / Jaki, Lena / Sallard, Erwan / Erhardt, Anja / Fuchs, Jonas / Ruzsics, Zsolt

Molecular therapy. Methods & clinical development

2024 Volume 32, Issue 2, Page(s) 101241

Abstract: While recombinant adenoviruses (rAds) are widely used in both laboratory and medical gene transfer, library-based applications using this vector platform are not readily available. Recently, we developed a new method, the CRISPR-Cas9 ... ...

Abstract	While recombinant adenoviruses (rAds) are widely used in both laboratory and medical gene transfer, library-based applications using this vector platform are not readily available. Recently, we developed a new method, the CRISPR-Cas9 mediated
Language	English
Publishing date	2024-03-18
Publishing country	United States
Document type	Journal Article
ZDB-ID	2872938-9
ISSN	2329-0501 ; 2329-0501
ISSN (online)	2329-0501
ISSN	2329-0501
DOI	10.1016/j.omtm.2024.101241
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Article: Rescue of Recombinant Adenoviruses by CRISPR/Cas-Mediated

Riedl, André / Fischer, Julian / Burgert, Hans-Gerhard / Ruzsics, Zsolt

Frontiers in microbiology

2022 Volume 13, Page(s) 854690

Abstract: Recombinant adenovirus (rAd) vectors represent one of the most frequently used vehicles for gene transfer ... ...

Abstract	Recombinant adenovirus (rAd) vectors represent one of the most frequently used vehicles for gene transfer applications
Language	English
Publishing date	2022-03-18
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2587354-4
ISSN	1664-302X
ISSN	1664-302X
DOI	10.3389/fmicb.2022.854690
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Article: Novel conditional plasmids regulated by chemical switches provide versatile tools for genetic engineering in Escherichia coli

Riedl, André / Gruber, Simone / Ruzsics, Zsolt

Plasmid. 2020 Sept., v. 111

2020

Abstract: Engineering bacterial genomes or foreign DNA cloned as bacterial artificial chromosomes (BACs) relies on usage of helper plasmids, which deliver the desired tools transiently into the bacteria to be modified. After the anticipated action is completed the ...

Abstract	Engineering bacterial genomes or foreign DNA cloned as bacterial artificial chromosomes (BACs) relies on usage of helper plasmids, which deliver the desired tools transiently into the bacteria to be modified. After the anticipated action is completed the helper plasmids need to be cured. To make this efficient, plasmids are used that are maintained by conditional amplicons or carry a counter-selection marker. Here, we describe new conditional plasmids that can be maintained or cured by using chemical induction or repression. Our method is based on the dependency of plasmids carrying ori6Kγ origin of replication on the presence of protein Π. Ori6Kγ based plasmids are tightly regulated conditional constructs, but they require usually special E. coli strains to operate. To avoid this, we placed the Π protein expression under the control of a co-expressed conditional repressor. Regulating the maintenance of plasmids with administration or removal of chemicals is fully compatible with any other conditional amplicons applied to date. Here, we describe methods for inducing sites specific recombination of BACs as an example. However, the same strategy might be used to construct appropriate helper plasmids for any other transient components of genome editing methodologies such as λred recombinases or CRISPR/Cas components.
Keywords	Escherichia coli ; genome ; plasmids ; protein synthesis ; recombinases ; replication origin
Language	English
Dates of publication	2020-09
Publishing place	Elsevier Inc.
Document type	Article
Note	NAL-AP-2-clean
ZDB-ID	282384-6
ISSN	1095-9890 ; 0147-619X
ISSN (online)	1095-9890
ISSN	0147-619X
DOI	10.1016/j.plasmid.2020.102531
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Cytomegaloviruses and Macrophages-Friends and Foes From Early on?

Baasch, Sebastian / Ruzsics, Zsolt / Henneke, Philipp

Frontiers in immunology

2020 Volume 11, Page(s) 793

Abstract: Starting at birth, newborn infants are exposed to numerous microorganisms. Adaptation of the innate immune system to them is a delicate process, with potentially advantageous and harmful implications for health development. Cytomegaloviruses (CMVs) are ... ...

Abstract	Starting at birth, newborn infants are exposed to numerous microorganisms. Adaptation of the innate immune system to them is a delicate process, with potentially advantageous and harmful implications for health development. Cytomegaloviruses (CMVs) are highly adapted to their specific mammalian hosts, with which they share millions of years of co-evolution. Throughout the history of mankind, human CMV has infected most infants in the first months of life without overt implications for health. Thus, CMV infections are intertwined with normal immune development. Nonetheless, CMV has retained substantial pathogenicity following infection
MeSH term(s)	Animals ; Cytomegalovirus/immunology ; Cytomegalovirus Infections/immunology ; Cytomegalovirus Infections/virology ; Host Adaptation/immunology ; Humans ; Immunity, Innate ; Immunity, Mucosal ; Immunomodulation ; Infant ; Infant, Newborn ; Macrophages/immunology ; Mice ; Monocytes/immunology
Language	English
Publishing date	2020-05-12
Publishing country	Switzerland
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Review
ZDB-ID	2606827-8
ISSN	1664-3224 ; 1664-3224
ISSN (online)	1664-3224
ISSN	1664-3224
DOI	10.3389/fimmu.2020.00793
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Article ; Online: Novel conditional plasmids regulated by chemical switches provide versatile tools for genetic engineering in Escherichia coli.

Riedl, André / Gruber, Simone / Ruzsics, Zsolt

Plasmid

2020 Volume 111, Page(s) 102531

Abstract	Engineering bacterial genomes or foreign DNA cloned as bacterial artificial chromosomes (BACs) relies on usage of helper plasmids, which deliver the desired tools transiently into the bacteria to be modified. After the anticipated action is completed the helper plasmids need to be cured. To make this efficient, plasmids are used that are maintained by conditional amplicons or carry a counter-selection marker. Here, we describe new conditional plasmids that can be maintained or cured by using chemical induction or repression. Our method is based on the dependency of plasmids carrying ori6Kγ origin of replication on the presence of protein Π. Ori6Kγ based plasmids are tightly regulated conditional constructs, but they require usually special E. coli strains to operate. To avoid this, we placed the Π protein expression under the control of a co-expressed conditional repressor. Regulating the maintenance of plasmids with administration or removal of chemicals is fully compatible with any other conditional amplicons applied to date. Here, we describe methods for inducing sites specific recombination of BACs as an example. However, the same strategy might be used to construct appropriate helper plasmids for any other transient components of genome editing methodologies such as λred recombinases or CRISPR/Cas components.
MeSH term(s)	Chromosomes, Artificial, Bacterial ; Chromosomes, Bacterial ; DNA Replication ; Escherichia coli/genetics ; Gene Editing ; Gene Expression Regulation, Bacterial ; Genetic Engineering ; Genome, Bacterial ; Plasmids/genetics ; Recombination, Genetic ; Temperature
Language	English
Publishing date	2020-09-10
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	282384-6
ISSN	1095-9890 ; 0147-619X
ISSN (online)	1095-9890
ISSN	0147-619X
DOI	10.1016/j.plasmid.2020.102531
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Zs.A 1736: Show issues

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Article ; Online: Suppression of adaptive NK cell expansion by macrophage-mediated phagocytosis inhibited by 2B4-CD48.

Li, Rui / Galindo, Cristian Camilo / Davidson, Dominique / Guo, Huaijian / Zhong, Ming-Chao / Qian, Jin / Li, Bin / Ruzsics, Zsolt / Lau, Colleen M / O'Sullivan, Timothy E / Vidal, Silvia M / Sun, Joseph C / Veillette, André

Cell reports

2024 Volume 43, Issue 3, Page(s) 113800

Abstract: Infection of mice by mouse cytomegalovirus (MCMV) triggers activation and expansion of ... ...

Abstract	Infection of mice by mouse cytomegalovirus (MCMV) triggers activation and expansion of Ly49H
MeSH term(s)	Animals ; Mice ; Receptors, Immunologic/metabolism ; CD48 Antigen/metabolism ; Antigens, CD/metabolism ; Lymphocyte Activation ; Killer Cells, Natural ; Receptors, Cell Surface/metabolism ; Carrier Proteins/metabolism ; Cytomegalovirus Infections ; Macrophages/metabolism ; Phagocytosis
Chemical Substances	Receptors, Immunologic ; CD48 Antigen ; Antigens, CD ; Receptors, Cell Surface ; Carrier Proteins
Language	English
Publishing date	2024-02-21
Publishing country	United States
Document type	Journal Article
ZDB-ID	2649101-1
ISSN	2211-1247 ; 2211-1247
ISSN (online)	2211-1247
ISSN	2211-1247
DOI	10.1016/j.celrep.2024.113800
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Article: TheraVision: Engineering platform technology for the development of oncolytic viruses based on herpes simplex virus type 1.

Funk, Christina / Uhlig, Nadja / Ruzsics, Zsolt / Baur, Florentin / Peindl, Matthias / Nietzer, Sarah / Epting, Karina / Vacun, Gabriele / Dandekar, Gudrun / Botteron, Catherine / Werno, Christian / Grunwald, Thomas / Bailer, Susanne M

Molecular therapy. Oncology

2024 Volume 32, Issue 1, Page(s) 200784

Abstract: Viruses are able to efficiently penetrate cells, multiply, and eventually kill infected cells, release tumor antigens, and activate the immune system. Therefore, viruses are highly attractive novel agents for cancer therapy. Clinical trials with first ... ...

Abstract	Viruses are able to efficiently penetrate cells, multiply, and eventually kill infected cells, release tumor antigens, and activate the immune system. Therefore, viruses are highly attractive novel agents for cancer therapy. Clinical trials with first generations of oncolytic viruses (OVs) are very promising but show significant need for optimization. The aim of TheraVision was to establish a broadly applicable engineering platform technology for combinatorial oncolytic virus and immunotherapy. Through genetic engineering, an attenuated herpes simplex virus type 1 (HSV1) was generated that showed increased safety compared to the wild-type strain. To demonstrate the modularity and the facilitated generation of new OVs, two transgenes encoding retargeting as well as immunomodulating single-chain variable fragments (scFvs) were integrated into the platform vector. The resulting virus selectively infected epidermal growth factor receptor (EGFR)-expressing cells and produced a functional immune checkpoint inhibitor against programmed cell death protein 1 (PD-1). Thus, both viral-mediated oncolysis and immune-cell-mediated therapy were combined into a single viral vector. Safety and functionality of the armed OVs have been shown in novel preclinical models ranging from patient-derived organoids and tissue-engineered human
Language	English
Publishing date	2024-02-28
Publishing country	United States
Document type	Journal Article
ISSN	2950-3299
ISSN	2950-3299
DOI	10.1016/j.omton.2024.200784
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Article: Evaluation of adenovirus 19a as a novel vector for mucosal vaccination against influenza A viruses

Lapuente, Dennis / Ruzsics, Zsolt / Tenbusch, Matthias / Thirion, Christian

Vaccine. 2018 May 03, v. 36, no. 19

2018

Abstract: Since preexisting immunity and enhanced infection rates in a clinical trial of an HIV vaccine have raised some concerns on adenovirus (Ad) serotype 5-based vaccines, we evaluated the subgroup D adenovirus serotype Ad19a for its suitability as novel viral ...

Abstract	Since preexisting immunity and enhanced infection rates in a clinical trial of an HIV vaccine have raised some concerns on adenovirus (Ad) serotype 5-based vaccines, we evaluated the subgroup D adenovirus serotype Ad19a for its suitability as novel viral vector vaccine against mucosal infections. In BALB/c mice, we compared the immunogenicity and efficacy of E1/E3-deleted Ad19a vectors encoding the influenza A virus (IAV)-derived antigens hemagglutinin (HA) and nucleoprotein (NP) to the most commonly used Ad5 vectors. The adenoviral vectors were applied intranasally and induced detectable antigen-specific T cell responses in the lung and in the spleen as well as robust antibody responses. A prior DNA immunization significantly improved the immunogenicity of both vectors and resulted in full protection against a lethal infection with a heterologous H3N2 virus. Nevertheless, the Ad5-based vectors were slightly superior in reducing viral replication in the lung which corresponded to higher NP-specific T cell responses measured in the lungs.
Keywords	Adenoviridae ; antibodies ; antigens ; clinical trials ; DNA ; hemagglutinins ; Human immunodeficiency virus ; immunogenicity ; Influenza A virus ; lungs ; mice ; nucleoproteins ; serotypes ; spleen ; T-lymphocytes ; vaccination ; vector vaccines ; virus replication
Language	English
Dates of publication	2018-0503
Size	p. 2712-2720.
Publishing place	Elsevier Ltd
Document type	Article
ZDB-ID	605674-x
ISSN	1873-2518 ; 0264-410X
ISSN (online)	1873-2518
ISSN	0264-410X
DOI	10.1016/j.vaccine.2018.02.075
Database	NAL-Catalogue (AGRICOLA)

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Article ; Online: Evaluation of adenovirus 19a as a novel vector for mucosal vaccination against influenza A viruses.

Lapuente, Dennis / Ruzsics, Zsolt / Thirion, Christian / Tenbusch, Matthias

Vaccine

2018 Volume 36, Issue 19, Page(s) 2712–2720

Abstract	Since preexisting immunity and enhanced infection rates in a clinical trial of an HIV vaccine have raised some concerns on adenovirus (Ad) serotype 5-based vaccines, we evaluated the subgroup D adenovirus serotype Ad19a for its suitability as novel viral vector vaccine against mucosal infections. In BALB/c mice, we compared the immunogenicity and efficacy of E1/E3-deleted Ad19a vectors encoding the influenza A virus (IAV)-derived antigens hemagglutinin (HA) and nucleoprotein (NP) to the most commonly used Ad5 vectors. The adenoviral vectors were applied intranasally and induced detectable antigen-specific T cell responses in the lung and in the spleen as well as robust antibody responses. A prior DNA immunization significantly improved the immunogenicity of both vectors and resulted in full protection against a lethal infection with a heterologous H3N2 virus. Nevertheless, the Ad5-based vectors were slightly superior in reducing viral replication in the lung which corresponded to higher NP-specific T cell responses measured in the lungs.
MeSH term(s)	A549 Cells ; Adenoviruses, Human/genetics ; Animals ; Female ; Gene Expression ; Genetic Vectors/immunology ; Humans ; Immunity, Humoral ; Immunity, Mucosal ; Influenza A Virus, H3N2 Subtype/pathogenicity ; Influenza A virus/pathogenicity ; Influenza Vaccines/immunology ; Influenza Vaccines/pharmacology ; Mice, Inbred BALB C ; Orthomyxoviridae Infections/prevention & control ; T-Lymphocytes/immunology ; Vaccines, DNA/immunology ; Vaccines, DNA/pharmacology
Chemical Substances	Influenza Vaccines ; Vaccines, DNA
Language	English
Publishing date	2018-04-05
Publishing country	Netherlands
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	605674-x
ISSN	1873-2518 ; 0264-410X
ISSN (online)	1873-2518
ISSN	0264-410X
DOI	10.1016/j.vaccine.2018.02.075
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Article ; Online: Neonatal immune challenge poses a sex-specific risk for epigenetic microglial reprogramming and behavioral impairment.

Schwabenland, Marius / Mossad, Omar / Sievert, Annika / Peres, Adam G / Ringel, Elena / Baasch, Sebastian / Kolter, Julia / Cascone, Giulia / Dokalis, Nikolaos / Vlachos, Andreas / Ruzsics, Zsolt / Henneke, Philipp / Prinz, Marco / Blank, Thomas

Nature communications

2023 Volume 14, Issue 1, Page(s) 2721

Abstract: While the precise processes underlying a sex bias in the development of central nervous system (CNS) disorders are unknown, there is growing evidence that an early life immune activation can contribute to the disease pathogenesis. When we mimicked an ... ...

Abstract	While the precise processes underlying a sex bias in the development of central nervous system (CNS) disorders are unknown, there is growing evidence that an early life immune activation can contribute to the disease pathogenesis. When we mimicked an early systemic viral infection or applied murine cytomegalovirus (MCMV) systemically in neonatal female and male mice, only male adolescent mice presented behavioral deficits, including reduced social behavior and cognition. This was paralleled by an increased amount of infiltrating T cells in the brain parenchyma, enhanced interferon-γ (IFNγ) signaling, and epigenetic reprogramming of microglial cells. These microglial cells showed increased phagocytic activity, which resulted in abnormal loss of excitatory synapses within the hippocampal brain region. None of these alterations were seen in female adolescent mice. Our findings underscore the early postnatal period's susceptibility to cause sex-dependent long-term CNS deficiencies following infections.
MeSH term(s)	Animals ; Female ; Male ; Mice ; Microglia/pathology ; Brain ; Central Nervous System Diseases/pathology ; Interferon-gamma/genetics ; Epigenesis, Genetic
Chemical Substances	Interferon-gamma (82115-62-6)
Language	English
Publishing date	2023-05-11
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2553671-0
ISSN	2041-1723 ; 2041-1723
ISSN (online)	2041-1723
ISSN	2041-1723
DOI	10.1038/s41467-023-38373-0
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