Controlling gene expression with the Q repressible binary expression system in Caenorhabditis elegans

We established a transcription-based binary gene expression system in Caenorhabditis elegans using the recently developed Q system. This system, derived from genes in Neurospora crassa, uses the transcriptional activator QF to induce the expression of target genes. Activation can be efficiently suppressed by the transcriptional repressor QS, and suppression can be relieved by the nontoxic small molecule quinic acid. We used QF, QS and quinic acid to achieve temporal and spatial control of transgene expression in various tissues in C. elegans. We also developed a split Q system, in which we separated QF into two parts encoding its DNA-binding and transcription-activation domains. Each domain showed negligible transcriptional activity when expressed alone, but expression of both reconstituted QF activity, providing additional combinatorial power to control gene expression.

[1]  Dana Carroll,et al.  Gene Activation Using FLP Recombinase in C. elegans , 2008, PLoS genetics.

[2]  K. Svoboda,et al.  Genetic Dissection of Neural Circuits , 2008, Neuron.

[3]  Erik M Jorgensen,et al.  Single-copy insertion of transgenes in Caenorhabditis elegans , 2008, Nature Genetics.

[4]  N. Munakata.  [Genetics of Caenorhabditis elegans]. , 1989, Tanpakushitsu kakusan koso. Protein, nucleic acid, enzyme.

[5]  M. Chalfie,et al.  Combinatorial Marking of Cells and Organelles with Reconstituted Fluorescent Proteins , 2004, Cell.

[6]  R. Porter,et al.  DNA transformation. , 1988, Methods in enzymology.

[7]  Oliver Hobert,et al.  A Toolkit and Robust Pipeline for the Generation of Fosmid-Based Reporter Genes in C. elegans , 2009, PloS one.

[8]  J. Barry,et al.  cis regulatory requirements for hypodermal cell-specific expression of the Caenorhabditis elegans cuticle collagen gene dpy-7 , 1997, Molecular and cellular biology.

[9]  C. Kenyon,et al.  MIG-13 Positions Migrating Cells along the Anteroposterior Body Axis of C. elegans , 1999, Cell.

[10]  D. Baillie,et al.  Molecular cloning and characterization of the dpy-20 gene of Caenorhabditis elegans , 1995, Molecular and General Genetics MGG.

[11]  Evan Z. Macosko,et al.  A huband-spoke circuit drives pheromone attraction and social behaviour in C . elegans , 2009 .

[12]  M. Case,et al.  The Wilhelmine E. Key 1989 invitational lecture. Organization and regulation of the qa (quinic acid) genes in Neurospora crassa and other fungi. , 1991, The Journal of heredity.

[13]  M. Gossen,et al.  Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. , 1992, Proceedings of the National Academy of Sciences of the United States of America.

[14]  John Tyler Bonner,et al.  Morphogenesis , 1965, The Physics of Living Matter: Space, Time and Information.

[15]  N. Perrimon,et al.  Targeted gene expression as a means of altering cell fates and generating dominant phenotypes. , 1993, Development.

[16]  D M Miller,et al.  Expression of the unc-4 homeoprotein in Caenorhabditis elegans motor neurons specifies presynaptic input. , 1995, Development.

[17]  L. Regan,et al.  Antiparallel Leucine Zipper-Directed Protein Reassembly: Application to the Green Fluorescent Protein , 2000 .

[18]  L. Nollet.  Food analysis by HPLC , 2012 .

[19]  M. Chalfie,et al.  Targeted cell killing by reconstituted caspases , 2007, Proceedings of the National Academy of Sciences.

[20]  Haojiang Luan,et al.  Refined Spatial Manipulation of Neuronal Function by Combinatorial Restriction of Transgene Expression , 2006, Neuron.

[21]  S. Shaham,et al.  Temporal Control of Cell-Specific Transgene Expression in Caenorhabditis elegans , 2007, Genetics.

[22]  Andrew Fire,et al.  Chapter 19 DNA Transformation , 1995 .

[23]  A. Fire,et al.  Recognition and silencing of repeated DNA. , 2000, Annual review of genetics.

[24]  J. Spieth,et al.  Operons in C. elegans: Polycistronic mRNA precursors are processed by trans-splicing of SL2 to downstream coding regions , 1993, Cell.

[25]  A. Fire,et al.  Soma-germline asymmetry in the distributions of embryonic RNAs in Caenorhabditis elegans. , 1994, Development.

[26]  Liqun Luo,et al.  Mosaic Analysis with a Repressible Cell Marker for Studies of Gene Function in Neuronal Morphogenesis , 1999, Neuron.

[27]  Liang Liang,et al.  The Q System: A Repressible Binary System for Transgene Expression, Lineage Tracing, and Mosaic Analysis , 2010, Cell.

[28]  N. Pei.  Application of Green Fluorescent Protein , 2001 .

[29]  S. Brenner,et al.  The structure of the nervous system of the nematode Caenorhabditis elegans. , 1986, Philosophical transactions of the Royal Society of London. Series B, Biological sciences.