Faithful Expression of Multiple Proteins via 2A-Peptide Self-Processing: A Versatile and Reliable Method for Manipulating Brain Circuits

A method allowing for efficient and quantitative coexpression of multiple heterologous proteins in neurons in vivo would be highly valuable for many applications in neuroscience. To date, different approaches, such as internal ribosomal entry site (IRES) elements ([Douin et al., 2004][1]),

[1]  Markus Affolter,et al.  Structural basis of BMP signalling inhibition by the cystine knot protein Noggin , 2002, Nature.

[2]  R. Kotin,et al.  Immunological Aspects of Recombinant Adeno-Associated Virus Delivery to the Mammalian Brain , 2002, Journal of Virology.

[3]  Philippe Soriano Generalized lacZ expression with the ROSA26 Cre reporter strain , 1999, Nature Genetics.

[4]  M. Gossen,et al.  Co-regulation of two gene activities by tetracycline via a bidirectional promoter. , 1995, Nucleic acids research.

[5]  G. Brewer,et al.  Optimized survival of hippocampal neurons in B27‐supplemented neurobasal™, a new serum‐free medium combination , 1993, Journal of neuroscience research.

[6]  R. Malinow,et al.  Postsynaptic Density 95 controls AMPA Receptor Incorporation during Long-Term Potentiation and Experience-Driven Synaptic Plasticity , 2004, The Journal of Neuroscience.

[7]  H. Hauser,et al.  Composition and arrangement of genes define the strength of IRES-driven translation in bicistronic mRNAs. , 2001, Nucleic acids research.

[8]  M. Ryan,et al.  Targeting of Proteins Derived from Self‐Processing Polyproteins Containing Multiple Signal Sequences , 2004, Traffic.

[9]  Takeharu Nagai,et al.  Shift anticipated in DNA microarray market , 2002, Nature Biotechnology.

[10]  Jinhyung Kim,et al.  A Genetic Switch for Epilepsy in Adult Mice , 2004, The Journal of Neuroscience.

[11]  M. Ryan,et al.  Cleavage of foot-and-mouth disease virus polyprotein is mediated by residues located within a 19 amino acid sequence. , 1991, The Journal of general virology.

[12]  Roberto Malinow,et al.  Synaptic AMPA Receptor Plasticity and Behavior , 2009, Neuron.

[13]  R. Tsien,et al.  A monomeric red fluorescent protein , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[14]  S. Kügler,et al.  Promoters and serotypes: targeting of adeno‐associated virus vectors for gene transfer in the rat central nervous system in vitro and in vivo , 2005, Experimental physiology.

[15]  H. Büeler,et al.  Recombinant AAV vectors containing the foot and mouth disease virus 2A sequence confer efficient bicistronic gene expression in cultured cells and rat substantia nigra neurons , 2001, Gene Therapy.

[16]  Murtaza Z Mogri,et al.  Targeting and Readout Strategies for Fast Optical Neural Control In Vitro and In Vivo , 2007, The Journal of Neuroscience.

[17]  R. Sprengel,et al.  Protein fusions with the kanamycin resistance gene from transposon Tn5. , 1984, EMBO Journal.

[18]  Takeharu Nagai,et al.  Functional Fluorescent Ca2+ Indicator Proteins in Transgenic Mice under TET Control , 2004, PLoS biology.

[19]  Y. Humeau,et al.  Dopamine gates LTP induction in lateral amygdala by suppressing feedforward inhibition , 2003, Nature Neuroscience.

[20]  D. Muller,et al.  A simple method for organotypic cultures of nervous tissue , 1991, Journal of Neuroscience Methods.

[21]  J. Lengler,et al.  FMDV-2A sequence and protein arrangement contribute to functionality of CYP2B1-reporter fusion protein. , 2005, Analytical biochemistry.

[22]  D Gani,et al.  Analysis of the aphthovirus 2A/2B polyprotein 'cleavage' mechanism indicates not a proteolytic reaction, but a novel translational effect: a putative ribosomal 'skip'. , 2001, The Journal of general virology.

[23]  B. Byrne,et al.  Recombinant adeno-associated virus purification using novel methods improves infectious titer and yield , 1999, Gene Therapy.

[24]  N. Nakatsuji,et al.  Efficient Multicistronic Expression of a Transgene in Human Embryonic Stem Cells , 2007, Stem cells.

[25]  P. Seeburg,et al.  Codon‐improved Cre recombinase (iCre) expression in the mouse , 2002, Genesis.

[26]  Takeharu Nagai,et al.  Cyan-emitting and orange-emitting fluorescent proteins as a donor/acceptor pair for fluorescence resonance energy transfer. , 2004, The Biochemical journal.

[27]  Pavel Osten,et al.  Stereotaxic gene delivery in the rodent brain , 2007, Nature Protocols.

[28]  K. Deisseroth,et al.  eNpHR: a Natronomonas halorhodopsin enhanced for optogenetic applications , 2008, Brain cell biology.

[29]  Claire Halpin,et al.  E unum pluribus: multiple proteins from a self-processing polyprotein. , 2006, Trends in biotechnology.

[30]  B. Dalby,et al.  Advanced transfection with Lipofectamine 2000 reagent: primary neurons, siRNA, and high-throughput applications. , 2004, Methods.

[31]  L. Créancier,et al.  Use and comparison of different internal ribosomal entry sites (IRES) in tricistronic retroviral vectors , 2004, BMC biotechnology.