Cell type–specific manipulation with GFP-dependent Cre recombinase
نویسندگان
چکیده
منابع مشابه
Stringent doxycycline dependent control of CRE recombinase in vivo.
The strategy of modulating gene activities in vivo via CRE/loxP recombination would greatly profit from subjecting the recombination event to an independent and stringent temporal control. Here, we describe a transgenic mouse line, LC-1, where the expression of the cre and luciferase gene is tightly controlled by the Tet system. Using the R26R mouse line as indicator for CRE activity, and mouse...
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Despite rapid advances in genome engineering technologies, inserting genes into precise locations in the human genome remains an outstanding problem. It has been suggested that site-specific recombinases can be adapted towards use as transgene delivery vectors. The specificity of recombinases can be altered either with directed evolution or via fusions to modular DNA-binding domains. Unfortunat...
متن کاملGenomic targeting with purified Cre recombinase.
Purified Cre recombinase protein introduced directly into cultured mammalian cells by lipofection catalyzes both site-specific chromosomal integration of a co-transfected lox targeting vector and precise excision of genomic DNA flanked by directly repeated lox sites. This procedure eliminates the need to transfect cre expression plasmids to activate recombination at lox sites. We used this simp...
متن کاملPreparation of cell-permeable Cre recombinase by expressed protein ligation
BACKGROUND Protein transduction is safer than viral vector-mediated transduction for the delivery of a therapeutic protein into a cell. Fusion proteins with an arginine-rich cell-penetrating peptide have been produced in E. coli, but the low solubility of the fusion protein expressed in E. coli impedes the large-scale production of fusion proteins from E. coli. RESULTS Expressed protein ligat...
متن کاملCRE recombinase-based positive-negative selection systems for genetic manipulation in Trypanosoma brucei.
The limited repertoire of drug-resistance markers imposes a serious obstacle to genetic manipulation of Trypanosoma brucei. Here we describe experiments with a fusion protein that allows positive selection for genome integration followed by CRE recombinase-mediated excision of the marker cassette that can be selected by ganciclovir, although the excision event is so efficient that selection is ...
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ژورنال
عنوان ژورنال: Nature Neuroscience
سال: 2015
ISSN: 1097-6256,1546-1726
DOI: 10.1038/nn.4081