LIVIVO - The Search Portal for Life Sciences

zur deutschen Oberfläche wechseln
Advanced search

Search results

Result 1 - 3 of total 3

Search options

  1. Article ; Online: Repurposing the Homing Endonuclease I-SceI for Positive Selection and Development of Gene-Editing Technologies.

    Lee, Kok Zhi / Mechikoff, Michael A / Parasa, Mrugesh Krishna / Rankin, Tyler J / Pandolfi, Paula / Fitzgerald, Kevin S / Hillman, Ethan T / Solomon, Kevin V

    ACS synthetic biology

    2022  Volume 11, Issue 1, Page(s) 53–60

    Abstract: Prokaryote genomes encode diverse programmable DNA endonucleases with significant potential for biotechnology and gene editing. However, these endonucleases differ significantly in their properties, which must be screened and measured. While positive ... ...

    Abstract Prokaryote genomes encode diverse programmable DNA endonucleases with significant potential for biotechnology and gene editing. However, these endonucleases differ significantly in their properties, which must be screened and measured. While positive selection screens based on ccdB and barnase have been developed to evaluate such proteins, their high levels of toxicity make them challenging to use. Here, we develop and validate a more robust positive selection screen based on the homing endonuclease I-SceI. Candidate endonucleases target and cure the I-SceI expression plasmid preventing induction of I-SceI-mediated double strand DNA breaks that lead to cell death in
    MeSH term(s) Deoxyribonuclease I ; Deoxyribonucleases, Type II Site-Specific/genetics ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Gene Editing ; Saccharomyces cerevisiae Proteins/genetics
    Chemical Substances Saccharomyces cerevisiae Proteins ; Deoxyribonuclease I (EC 3.1.21.1) ; Deoxyribonucleases, Type II Site-Specific (EC 3.1.21.4)
    Language English
    Publishing date 2022-01-10
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
    ISSN 2161-5063
    ISSN (online) 2161-5063
    DOI 10.1021/acssynbio.1c00340
    Database MEDical Literature Analysis and Retrieval System OnLINE

    More links

    Kategorien

    Order via subito

    This service is chargeable due to the Delivery terms set by subito. Orders including an article and supplementary material will be classified as separate orders. In these cases, fees will be demanded for each order.

    Inter-library loan at ZB MED

    Your chosen title can be delivered directly to ZB MED Cologne location if you are registered as a user at ZB MED Cologne.

  2. Article ; Online: Degradation of hydrogel beads for the detection of serum bicarbonate levels for the diagnosis of metabolic alkalosis at the point of care.

    Pandolfi, Paula / Zhang, Hanxu / Nadalin, Yui E / Prasetyo, Matthew E / Toubian, Adam A / Wu, Benjamin M / Kamei, Daniel T

    Soft matter

    2022  Volume 18, Issue 42, Page(s) 8147–8156

    Abstract: In this work, we present a novel point-of-care hydrogel-based diagnostic device for the rapid detection of elevated bicarbonate levels in serum for the diagnosis of mild to severe cases of metabolic alkalosis. Our system consists of hydrogel beads ... ...

    Abstract In this work, we present a novel point-of-care hydrogel-based diagnostic device for the rapid detection of elevated bicarbonate levels in serum for the diagnosis of mild to severe cases of metabolic alkalosis. Our system consists of hydrogel beads composed of calcium alginate and the nonionic polymer dextran. This assay utilizes the reaction of sodium bicarbonate and citric acid to produce citrate, a metal chelator capable of competitively binding to calcium cations in the gel matrix to trigger hydrogel degradation. This results in successful detection of elevated bicarbonate concentrations in less than one hour. Specifically, critically high bicarbonate concentrations of 50, 45, and 40 mmol L
    MeSH term(s) Humans ; Bicarbonates ; Point-of-Care Systems ; Hydrogels ; Alkalosis ; Citric Acid ; Alginates
    Chemical Substances Bicarbonates ; Hydrogels ; Citric Acid (2968PHW8QP) ; Alginates
    Language English
    Publishing date 2022-11-02
    Publishing country England
    Document type Journal Article
    ZDB-ID 2191476-X
    ISSN 1744-6848 ; 1744-683X
    ISSN (online) 1744-6848
    ISSN 1744-683X
    DOI 10.1039/d2sm01014c
    Database MEDical Literature Analysis and Retrieval System OnLINE

    More links

    Kategorien

    Order via subito

    This service is chargeable due to the Delivery terms set by subito. Orders including an article and supplementary material will be classified as separate orders. In these cases, fees will be demanded for each order.

  3. Article ; Online: NgAgo possesses guided DNA nicking activity.

    Lee, Kok Zhi / Mechikoff, Michael A / Kikla, Archana / Liu, Arren / Pandolfi, Paula / Fitzgerald, Kevin / Gimble, Frederick S / Solomon, Kevin V

    Nucleic acids research

    2021  Volume 49, Issue 17, Page(s) 9926–9937

    Abstract: Prokaryotic Argonautes (pAgos) have been proposed as more flexible tools for gene-editing as they do not require sequence motifs adjacent to their targets for function, unlike popular CRISPR/Cas systems. One promising pAgo candidate, from the halophilic ... ...

    Abstract Prokaryotic Argonautes (pAgos) have been proposed as more flexible tools for gene-editing as they do not require sequence motifs adjacent to their targets for function, unlike popular CRISPR/Cas systems. One promising pAgo candidate, from the halophilic archaeon Natronobacterium gregoryi (NgAgo), has been the subject of debate regarding its potential in eukaryotic systems. Here, we revisit this enzyme and characterize its function in prokaryotes. NgAgo expresses poorly in non-halophilic hosts with most of the protein being insoluble and inactive even after refolding. However, we report that the soluble fraction does indeed act as a nicking DNA endonuclease. NgAgo shares canonical domains with other catalytically active pAgos but also contains a previously unrecognized single-stranded DNA binding domain (repA). Both repA and the canonical PIWI domains participate in DNA cleavage activities of NgAgo. NgAgo can be programmed with guides to nick targeted DNA in Escherichia coli and in vitro 1 nt outside the 3' end of the guide sequence. We also found that these endonuclease activities are essential for enhanced NgAgo-guided homologous recombination, or gene-editing, in E. coli. Collectively, our results demonstrate the potential of NgAgo for gene-editing and provide new insight into seemingly contradictory reports.
    MeSH term(s) Argonaute Proteins/metabolism ; DNA Cleavage ; DNA Helicases/genetics ; DNA, Bacterial/genetics ; DNA, Bacterial/metabolism ; Escherichia coli/genetics ; Gene Editing/methods ; Homologous Recombination/genetics ; Natronobacterium/enzymology ; Natronobacterium/genetics ; Natronobacterium/metabolism ; Trans-Activators/genetics
    Chemical Substances Argonaute Proteins ; DNA, Bacterial ; Trans-Activators ; replication initiator protein ; DNA Helicases (EC 3.6.4.-)
    Language English
    Publishing date 2021-09-03
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, U.S. Gov't, Non-P.H.S.
    ZDB-ID 186809-3
    ISSN 1362-4962 ; 1362-4954 ; 0301-5610 ; 0305-1048
    ISSN (online) 1362-4962 ; 1362-4954
    ISSN 0301-5610 ; 0305-1048
    DOI 10.1093/nar/gkab757
    Database MEDical Literature Analysis and Retrieval System OnLINE

    More links

    Kategorien

    In stock of ZB MED Cologne/Königswinter

    Zs.A 1067: Show issues Location:
    Je nach Verfügbarkeit (siehe Angabe bei Bestand)
    bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular
    Jg. 1995 - 2021: Lesesall (1.OG)
    ab Jg. 2022: Lesesaal (EG)

    Order via subito

    This service is chargeable due to the Delivery terms set by subito. Orders including an article and supplementary material will be classified as separate orders. In these cases, fees will be demanded for each order.

To top