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Artikel ; Online: A Dual-Function "TRE-Lox" System for Genetic Deletion or Reversible, Titratable, and Near-Complete Downregulation of Cathepsin D.

Terron, Heather M / Maranan, Derek S / Burgard, Luke A / LaFerla, Frank M / Lane, Shelley / Leissring, Malcolm A

International journal of molecular sciences

2023  Band 24, Heft 7

Abstract: Commonly employed methods for reversibly disrupting gene expression, such as those based on RNAi or CRISPRi, are rarely capable of achieving >80-90% downregulation, making them unsuitable for targeting genes that require more complete disruption to ... ...

Abstract Commonly employed methods for reversibly disrupting gene expression, such as those based on RNAi or CRISPRi, are rarely capable of achieving >80-90% downregulation, making them unsuitable for targeting genes that require more complete disruption to elicit a phenotype. Genetic deletion, on the other hand, while enabling complete disruption of target genes, often produces undesirable irreversible consequences such as cytotoxicity or cell death. Here we describe the design, development, and detailed characterization of a dual-function "TRE-Lox" system for effecting either (a) doxycycline (Dox)-mediated downregulation or (b) genetic deletion of a target gene-the lysosomal aspartyl protease cathepsin D (CatD)-based on targeted insertion of a tetracycline-response element (TRE) and two LoxP sites into the 5' end of the endogenous CatD gene (
Mesh-Begriff(e) Animals ; Mice ; Cathepsin D/genetics ; Cathepsin D/metabolism ; Down-Regulation/genetics ; Fibroblasts/metabolism ; Tetracycline ; Doxycycline/pharmacology ; Response Elements
Chemische Substanzen Cathepsin D (EC 3.4.23.5) ; Tetracycline (F8VB5M810T) ; Doxycycline (N12000U13O)
Sprache Englisch
Erscheinungsdatum 2023-04-04
Erscheinungsland Switzerland
Dokumenttyp Journal Article
ZDB-ID 2019364-6
ISSN 1422-0067 ; 1422-0067 ; 1661-6596
ISSN (online) 1422-0067
ISSN 1422-0067 ; 1661-6596
DOI 10.3390/ijms24076745
Datenquelle MEDical Literature Analysis and Retrieval System OnLINE

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