Abstract
Here, we present MultiSite Assembly of Gateway Induced Clones (MAGIC), which leverages Gateway-based recombinatorial cloning technology for rapid, modular assembly of plasmids to facilitate transgenesis in cells and vertebrate animal models. The MAGIC collection of plasmids spans a range of in vitro and in vivo uses, from tools for optically and chemically tunable gene expression, to simultaneous expression of microRNAs and fluorescent reporters, to a suite of distinct subcellular compartmental fluorescent reporters, to Cre and Dre recombinase-dependent gene expression. MAGIC system components are compatible with existing MultiSite Gateway Tol2 systems currently used in zebrafish and mammalian lentiviral and adenoviral Destination vectors, allowing rapid cross-species experimentation. The kit also includes novel vectors for stable transgene integration into host genomes in vitro or in vivo when used with piggyBac transposase, I-Sce meganuclease or Tol2 transposase. Collectively, the MAGIC system facilitates transgenesis in cultured mammalian cells, mouse, chick and zebrafish embryos, enabling the rapid generation of innovative DNA constructs for biological research due to a shared, common plasmid platform.
Keywords:
piggyBac; Cloning; Cre/lox; Dre/rox; Fluorescence; Gateway; LexA/LexOp; Tet/dox.