Nature Communications · 2020 · 65 citations · 19 references
Life SciencesBiotechnologyGeneticsGene ExpressionGenetic EngineeringGenome EngineeringOptogeneticsRecombinant Protein ExpressionRecombinant Protein ProductionTransgene ConstructionTransgene DesignTransgene Expression ControlBiomedical ApplicationsCre-lox Recombination SystemsMammalian Expression SystemsMammalian Expression VectorsMolecular TargetingOptogenetic Gene TherapySequence SimilarityViral Vector Production
Optogenetic genome engineering tools enable spatiotemporal control of gene expression and provide new insight into biological function. Here, we report the new version of genetically encoded photoactivatable (PA) Cre recombinase, PA-Cre 3.0. To improve PA-Cre technology, we compare light-dimerization tools and optimize for mammalian expression using a CAG promoter, Magnets, and 2A self-cleaving peptide. To prevent background recombination caused by the high sequence similarity in the dimerization domains, we modify the codons for mouse gene targeting and viral production. Overall, these modifications significantly reduce dark leak activity and improve blue-light induction developing our new version, PA-Cre 3.0. As a resource, we have generated and validated AAV-PA-Cre 3.0 as well as two mouse lines that can conditionally express PA-Cre 3.0. Together these new tools will facilitate further biological and biomedical research.
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A "Silent" Polymorphism in the <i>MDR</i> 1 Gene Changes Substrate Specificity
Chava Kimchi‐Sarfaty, Jung Mi Oh, In‐Wha Kim et al. · Science · 2006 · 2.5K citations · Full text
Gurkan Guntas, Ryan A. Hallett, Seth P. Zimmerman et al. · Proceedings of the National Academy of Sciences · 2014 · 704 citations · Full text