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Article ; Online: Current and Prospective Applications of CRISPR-Cas12a in Pluricellular Organisms.

Khan, Shaheen / Sallard, Erwan

2022 Volume 65, Issue 2, Page(s) 196–205

Abstract: CRISPR-Cas systems play a critical role in the prokaryotic adaptive immunity against mobile genetic elements, such as phages and foreign plasmids. In the last decade, Cas9 has been established as a powerful and versatile gene editing tool. In its wake, ... ...

Abstract	CRISPR-Cas systems play a critical role in the prokaryotic adaptive immunity against mobile genetic elements, such as phages and foreign plasmids. In the last decade, Cas9 has been established as a powerful and versatile gene editing tool. In its wake, the novel RNA-guided endonuclease system CRISPR-Cas12a is transforming biological research due to its unique properties, such as its high specificity or its ability to target T-rich motifs, to induce staggered double-strand breaks and to process RNA arrays. Meanwhile, there is an increasing need for efficient and safe gene activation, repression or editing in pluricellular organisms for crop improvement, gene therapy, research model development, and other goals. In this article, we review CRISPR-Cas12a applications in pluricellular organisms and discuss how the challenges characteristic of these complex models, such as vectorization or temperature variations in ectothermic species, can be overcome.
MeSH term(s)	CRISPR-Cas Systems ; Gene Editing ; Endonucleases/metabolism ; Plasmids ; RNA
Chemical Substances	Endonucleases (EC 3.1.-) ; RNA (63231-63-0)
Language	English
Publishing date	2022-08-08
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	1193057-3
ISSN	1559-0305 ; 1073-6085
ISSN (online)	1559-0305
ISSN	1073-6085
DOI	10.1007/s12033-022-00538-5
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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 ; Online: Current and Prospective Applications of CRISPR-Cas12a in Pluricellular Organisms

Khan, Shaheen / Sallard, Erwan

Mol Biotechnol. 2023 Feb., v. 65, no. 2 p.196-205

2023

Abstract	CRISPR-Cas systems play a critical role in the prokaryotic adaptive immunity against mobile genetic elements, such as phages and foreign plasmids. In the last decade, Cas9 has been established as a powerful and versatile gene editing tool. In its wake, the novel RNA-guided endonuclease system CRISPR-Cas12a is transforming biological research due to its unique properties, such as its high specificity or its ability to target T-rich motifs, to induce staggered double-strand breaks and to process RNA arrays. Meanwhile, there is an increasing need for efficient and safe gene activation, repression or editing in pluricellular organisms for crop improvement, gene therapy, research model development, and other goals. In this article, we review CRISPR-Cas12a applications in pluricellular organisms and discuss how the challenges characteristic of these complex models, such as vectorization or temperature variations in ectothermic species, can be overcome.
Keywords	CRISPR-Cas systems ; RNA ; adaptive immunity ; ectothermy ; gene activation ; gene therapy ; genes ; plasmids ; temperature
Language	English
Dates of publication	2023-02
Size	p. 196-205.
Publishing place	Springer US
Document type	Article ; Online
Note	Review
ZDB-ID	1193057-3
ISSN	1559-0305 ; 1073-6085
ISSN (online)	1559-0305
ISSN	1073-6085
DOI	10.1007/s12033-022-00538-5
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Article ; Online: The Adenovirus Vector Platform: Novel Insights into Rational Vector Design and Lessons Learned from the COVID-19 Vaccine.

Sallard, Erwan / Zhang, Wenli / Aydin, Malik / Schröer, Katrin / Ehrhardt, Anja

Viruses

2023 Volume 15, Issue 1

Abstract: The adenovirus vector platform remains one of the most efficient toolboxes for generation of transfer vehicles used in gene therapy and virotherapy to treat tumors, as well as vaccines to protect from infectious diseases. The adenovirus genome and ... ...

Abstract	The adenovirus vector platform remains one of the most efficient toolboxes for generation of transfer vehicles used in gene therapy and virotherapy to treat tumors, as well as vaccines to protect from infectious diseases. The adenovirus genome and capsids can be modified using highly efficient techniques, and vectors can be produced at high titers, which facilitates their rapid adaptation to current needs and disease applications. Over recent years, the adenovirus vector platform has been in the center of attention for vaccine development against the ongoing coronavirus SARS-CoV-2/COVID-19 pandemic. The worldwide deployment of these vaccines has greatly deepened the knowledge on virus-host interactions and highlighted the need to further improve the effectiveness and safety not only of adenovirus-based vaccines but also of gene therapy and oncolytic virotherapy vectors. Based on the current evidence, we discuss here how adenoviral vectors can be further improved by intelligent molecular design. This review covers the full spectrum of state-of-the-art strategies to avoid vector-induced side effects ranging from the vectorization of non-canonical adenovirus types to novel genome engineering techniques.
MeSH term(s)	Humans ; COVID-19 Vaccines ; Pandemics ; COVID-19/prevention & control ; SARS-CoV-2/genetics ; Adenoviridae/genetics ; Vaccines ; Genetic Vectors/genetics
Chemical Substances	COVID-19 Vaccines ; Vaccines
Language	English
Publishing date	2023-01-11
Publishing country	Switzerland
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't
ZDB-ID	2516098-9
ISSN	1999-4915 ; 1999-4915
ISSN (online)	1999-4915
ISSN	1999-4915
DOI	10.3390/v15010204
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Article ; Online: Electrophysiology of ionotropic GABA receptors.

Sallard, Erwan / Letourneur, Diane / Legendre, Pascal

Cellular and molecular life sciences : CMLS

2021 Volume 78, Issue 13, Page(s) 5341–5370

Abstract: ... ...

Abstract	GABA
MeSH term(s)	Animals ; Electrophysiology ; Humans ; Neurons/cytology ; Neurons/physiology ; Receptors, GABA-A/metabolism ; gamma-Aminobutyric Acid/metabolism
Chemical Substances	Receptors, GABA-A ; gamma-Aminobutyric Acid (56-12-2)
Language	English
Publishing date	2021-06-01
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	1358415-7
ISSN	1420-9071 ; 1420-682X
ISSN (online)	1420-9071
ISSN	1420-682X
DOI	10.1007/s00018-021-03846-2
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Book ; Online: jvanheld/SARS-CoV-2_origins

Jacques van Helden / Erwan Sallard

MedecineSciences-preprint

2020

Abstract: Version of the code used to generate the results and figures of the article "Retrouver les origines du SARS-COV-2 dans les phylogénies de coronavirus", to be published in the Aug-Sept 2020 issue of the journal Médecine & Sciences. The article has also ... ...

Abstract	Version of the code used to generate the results and figures of the article "Retrouver les origines du SARS-COV-2 dans les phylogénies de coronavirus", to be published in the Aug-Sept 2020 issue of the journal Médecine & Sciences. The article has also been fully translated to English under the title "Tracing the origins of SARS-COV-2 in coronavirus phylogenies", that will become available in HAL (https://hal.archives-ouvertes.fr/).
Keywords	covid19
Publishing date	2020-07-06
Publishing country	eu
Document type	Book ; Online
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Article: Electrophysiology of ionotropic GABA receptors

Sallard, Erwan / Letourneur, Diane / Legendre, Pascal

Cellular and molecular life sciences. 2021 July, v. 78, no. 13

2021

Abstract: GABAA receptors are ligand-gated chloride channels and ionotropic receptors of GABA, the main inhibitory neurotransmitter in vertebrates. In this review, we discuss the major and diverse roles GABAA receptors play in the regulation of neuronal ... ...

Abstract	GABAA receptors are ligand-gated chloride channels and ionotropic receptors of GABA, the main inhibitory neurotransmitter in vertebrates. In this review, we discuss the major and diverse roles GABAA receptors play in the regulation of neuronal communication and the functioning of the brain. GABAA receptors have complex electrophysiological properties that enable them to mediate different types of currents such as phasic and tonic inhibitory currents. Their activity is finely regulated by membrane voltage, phosphorylation and several ions. GABAA receptors are pentameric and are assembled from a diverse set of subunits. They are subdivided into numerous subtypes, which differ widely in expression patterns, distribution and electrical activity. Substantial variations in macroscopic neural behavior can emerge from minor differences in structure and molecular activity between subtypes. Therefore, the diversity of GABAA receptors widens the neuronal repertoire of responses to external signals and contributes to shaping the electrical activity of neurons and other cell types.
Keywords	brain ; chlorides ; electric potential difference ; electrophysiology ; neurons ; neurotransmitters ; phosphorylation
Language	English
Dates of publication	2021-07
Size	p. 5341-5370.
Publishing place	Springer International Publishing
Document type	Article
Note	Review
ZDB-ID	1358415-7
ISSN	1420-9071 ; 1420-682X
ISSN (online)	1420-9071
ISSN	1420-682X
DOI	10.1007/s00018-021-03846-2
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Article: Tracing the origins of SARS-COV-2 in coronavirus phylogenies: a review.

Sallard, Erwan / Halloy, José / Casane, Didier / Decroly, Etienne / van Helden, Jacques

Environmental chemistry letters

2021 Volume 19, Issue 2, Page(s) 769–785

Abstract: SARS-CoV-2 is a new human coronavirus (CoV), which emerged in China in late 2019 and is responsible for the global COVID-19 pandemic that caused more than 97 million infections and 2 million deaths in 12 months. Understanding the origin of this virus is ... ...

Abstract	SARS-CoV-2 is a new human coronavirus (CoV), which emerged in China in late 2019 and is responsible for the global COVID-19 pandemic that caused more than 97 million infections and 2 million deaths in 12 months. Understanding the origin of this virus is an important issue, and it is necessary to determine the mechanisms of viral dissemination in order to contain future epidemics. Based on phylogenetic inferences, sequence analysis and structure-function relationships of coronavirus proteins, informed by the knowledge currently available on the virus, we discuss the different scenarios on the origin-natural or synthetic-of the virus. The data currently available are not sufficient to firmly assert whether SARS-CoV2 results from a zoonotic emergence or from an accidental escape of a laboratory strain. This question needs to be solved because it has important consequences on the risk/benefit balance of our interactions with ecosystems, on intensive breeding of wild and domestic animals, on some laboratory practices and on scientific policy and biosafety regulations. Regardless of COVID-19 origin, studying the evolution of the molecular mechanisms involved in the emergence of pandemic viruses is essential to develop therapeutic and vaccine strategies and to prevent future zoonoses. This article is a translation and update of a French article published in Médecine/Sciences, August/September 2020 (10.1051/medsci/2020123). Supplementary information: The online version of this article (10.1007/s10311-020-01151-1) contains supplementary material, which is available to authorized users.
Language	English
Publishing date	2021-02-04
Publishing country	United States
Document type	Journal Article ; Review
ZDB-ID	2107984-5
ISSN	1610-3661 ; 1610-3653
ISSN (online)	1610-3661
ISSN	1610-3653
DOI	10.1007/s10311-020-01151-1
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Article: Simplified Point-of-Care Full SARS-CoV-2 Genome Sequencing Using Nanopore Technology.

Pembaur, Anton / Sallard, Erwan / Weil, Patrick Philipp / Ortelt, Jennifer / Ahmad-Nejad, Parviz / Postberg, Jan

Microorganisms

2021 Volume 9, Issue 12

Abstract: The scale of the ongoing SARS-CoV-2 pandemic warrants the urgent establishment of a global decentralized surveillance system to recognize local outbreaks and the emergence of novel variants of concern. Among available deep-sequencing technologies, ... ...

Abstract	The scale of the ongoing SARS-CoV-2 pandemic warrants the urgent establishment of a global decentralized surveillance system to recognize local outbreaks and the emergence of novel variants of concern. Among available deep-sequencing technologies, nanopore-sequencing could be an important cornerstone, as it is mobile, scalable, and cost-effective. Therefore, streamlined nanopore-sequencing protocols need to be developed and optimized for SARS-CoV-2 variants identification. We adapted and simplified existing workflows using the 'midnight' 1200 bp amplicon split primer sets for PCR, which produce tiled overlapping amplicons covering almost the entire SARS-CoV-2 genome. Subsequently, we applied Oxford Nanopore Rapid Barcoding and the portable MinION Mk1C sequencer combined with the interARTIC bioinformatics pipeline. We tested a simplified and less time-consuming workflow using SARS-CoV-2-positive specimens from clinical routine and identified the CT value as a useful pre-analytical parameter, which may help to decrease sequencing failures rates. Complete pipeline duration was approx. 7 h for one specimen and approx. 11 h for 12 multiplexed barcoded specimens. The adapted protocol contains fewer processing steps and can be completely conducted within one working day. Diagnostic CT values deduced from qPCR standardization experiments can act as principal criteria for specimen selection. As a guideline, SARS-CoV-2 genome copy numbers lower than 4 × 10
Language	English
Publishing date	2021-12-16
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	2720891-6
ISSN	2076-2607
ISSN	2076-2607
DOI	10.3390/microorganisms9122598
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Article ; Online: Novel Group C Oncolytic Adenoviruses Carrying a miRNA Inhibitor Demonstrate Enhanced Oncolytic Activity In Vitro and In Vivo.

Doerner, Johannes / Sallard, Erwan / Zhang, Wenli / Solanki, Manish / Liu, Jing / Ehrke-Schulz, Eric / Zirngibl, Hubert / Lieber, André / Ehrhardt, Anja

Molecular cancer therapeutics

2022 Volume 21, Issue 3, Page(s) 460–470

Abstract: Oncolytic adenoviruses (OAd) represent an attractive treatment option for cancer. Clinical efficacy of commonly utilized human adenovirus type 5 (Ad5)-based oncolytic viruses is limited by variable expression levels of the coxsackie- and adenovirus ... ...

Abstract	Oncolytic adenoviruses (OAd) represent an attractive treatment option for cancer. Clinical efficacy of commonly utilized human adenovirus type 5 (Ad5)-based oncolytic viruses is limited by variable expression levels of the coxsackie- and adenovirus receptor (CAR) in tumor cells and high prevalence of neutralizing antibodies against human Ad5. However, previous studies have highlighted alternative human Ad types as promising candidates for oncolytic therapy. In this study, we generated novel OAds based on Ad1, -2, -5, and -6 derived from species C Ads. These OAds contain a 24-bp deletion in the early gene E1A for tumor selective replication and express the RNAi inhibitor P19. We examined these OAds for in vitro anticancer activity on various cancer cell lines derived from lung, colon, gynecologic, bone, and pancreatic carcinoma. In most surveyed cell lines, OAds based on Ad1, -2, and -6 demonstrated higher cell lysis capability compared with Ad5, suggesting enhanced oncolytic potential. Moreover, enhanced oncolytic activity was associated with P19 expression in a cell type-dependent manner. We further explored a A549 tumor xenograft mouse model to compare the novel OAds directly with Ad5 and H101, an oncolytic adenovirus used in clinical trials. These P19-containing OAds based on Ad1, -2, and -6 showed significantly decelerated tumor progression compared with H101, indicating better antitumor potency in vivo. Our studies provide a novel path for OAd development based on alternative Ad types with improved effectiveness by RNA interference suppression.
MeSH term(s)	Adenoviridae/genetics ; Animals ; Cell Line, Tumor ; Female ; Genetic Vectors ; Humans ; Mice ; MicroRNAs/metabolism ; Oncolytic Virotherapy ; Oncolytic Viruses/genetics ; Oncolytic Viruses/metabolism ; Virus Replication ; Xenograft Model Antitumor Assays
Chemical Substances	MicroRNAs
Language	English
Publishing date	2022-01-13
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2063563-1
ISSN	1538-8514 ; 1535-7163
ISSN (online)	1538-8514
ISSN	1535-7163
DOI	10.1158/1535-7163.MCT-21-0240
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