Article ; Online: A Dual-Function "TRE-Lox" System for Genetic Deletion or Reversible, Titratable, and Near-Complete Downregulation of Cathepsin D.
International journal of molecular sciences
2023 Volume 24, Issue 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 ( |
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| MeSH term(s) | Animals ; Mice ; Cathepsin D/genetics ; Cathepsin D/metabolism ; Down-Regulation/genetics ; Fibroblasts/metabolism ; Tetracycline ; Doxycycline/pharmacology ; Response Elements |
| Chemical Substances | Cathepsin D (EC 3.4.23.5) ; Tetracycline (F8VB5M810T) ; Doxycycline (N12000U13O) |
| Language | English |
| Publishing date | 2023-04-04 |
| Publishing country | Switzerland |
| Document type | 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 |
| Database | MEDical Literature Analysis and Retrieval System OnLINE |
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