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Genetics, Vol. 157, 1217-1226, March 2001, Copyright © 2001

Creation of Low-Copy Integrated Transgenic Lines in Caenorhabditis elegans

Vida Praitisa, Elizabeth Caseya, David Collara, and Judith Austina
a Department of Molecular Genetics and Cell Biology, University of Chicago, Chicago, Illinois 60637

Corresponding author: Judith Austin, Department of Molecular Genetics and Cell Biology, University of Chicago, 920 E. 58th St. Chicago, IL 60637, jaustin{at}midway.uchicago.edu (E-mail)

Communicating editor: B. J. MEYER

In Caenorhabditis elegans, transgenic lines are typically created by injecting DNA into the hermaphrodite germline to form multicopy extrachromosomal DNA arrays. This technique is a reliable means of expressing transgenes in C. elegans, but its use has limitations. Because extrachromosomal arrays are semistable, only a fraction of the animals in a transgenic extrachromosomal array line are transformed. In addition, because extrachromosomal arrays can contain hundreds of copies of the transforming DNA, transgenes may be overexpressed, misexpressed, or silenced. We have developed an alternative method for C. elegans transformation, using microparticle bombardment, that produces single- and low-copy chromosomal insertions. Using this method, we find that it is possible to create integrated transgenic lines that reproducibly express GFP reporter constructs without the variations in expression level and pattern frequently exhibited by extrachromosomal array lines. In addition, we find that low-copy integrated lines can also be used to express transgenes in the C. elegans germline, where conventional extrachromosomal arrays typically fail to express due to germline silencing.




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J. Novelli, A. P. Page, and J. Hodgkin
The C Terminus of Collagen SQT-3 Has Complex and Essential Functions in Nematode Collagen Assembly
Genetics, April 1, 2006; 172(4): 2253 - 2267.
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 Mol. Biol. CellHome page
C. C.-H. Chen, P. J. Schweinsberg, S. Vashist, D. P. Mareiniss, E. J. Lambie, and B. D. Grant
RAB-10 Is Required for Endocytic Recycling in the Caenorhabditis elegans Intestine
Mol. Biol. Cell, March 1, 2006; 17(3): 1286 - 1297.
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 Proc. Natl. Acad. Sci. USAHome page
Z. Bao, J. I. Murray, T. Boyle, S. L. Ooi, M. J. Sandel, and R. H. Waterston
Automated cell lineage tracing in Caenorhabditis elegans
PNAS, February 21, 2006; 103(8): 2707 - 2712.
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 DevelopmentHome page
H. Liu, T. J. Strauss, M. B. Potts, and S. Cameron
Direct regulation of egl-1 and of programmed cell death by the Hox protein MAB-5 and by CEH-20, a C. elegans homolog of Pbx1
Development, February 15, 2006; 133(4): 641 - 650.
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 DevelopmentHome page
A. E. Burrows, B. K. Sceurman, M. E. Kosinski, C. T. Richie, P. L. Sadler, J. M. Schumacher, and A. Golden
The C. elegans Myt1 ortholog is required for the proper timing of oocyte maturation
Development, February 15, 2006; 133(4): 697 - 709.
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 Nucleic Acids ResHome page
C. T. Dolphin and I. A. Hope
Caenorhabditis elegans reporter fusion genes generated by seamless modification of large genomic DNA clones.
Nucleic Acids Res., January 1, 2006; 34(9): e72 - e72.
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 Mol. Biol. CellHome page
J. M. Squirrell, Z. T. Eggers, N. Luedke, B. Saari, A. Grimson, G. E. Lyons, P. Anderson, and J. G. White
CAR-1, a Protein That Localizes with the mRNA Decapping Component DCAP-1, Is Required for Cytokinesis and ER Organization in Caenorhabditis elegans Embryos
Mol. Biol. Cell, January 1, 2006; 17(1): 336 - 344.
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 Genes Dev.Home page
E. Martinez-Perez and A. M. Villeneuve
HTP-1-dependent constraints coordinate homolog pairing and synapsis and promote chiasma formation during C. elegans meiosis
Genes & Dev., November 15, 2005; 19(22): 2727 - 2743.
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 Nucleic Acids ResHome page
H. E. Sassi, S. Renihan, A. M. Spence, and R. L. Cooperstock
Gene CATCHR--Gene Cloning And Tagging for Caenorhabditis elegans using yeast Homologous Recombination: a novel approach for the analysis of gene expression
Nucleic Acids Res., October 27, 2005; 33(18): e163 - e163.
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 JCBHome page
A. Audhya, F. Hyndman, I. X. McLeod, A. S. Maddox, J. R. Yates III, A. Desai, and K. Oegema
A complex containing the Sm protein CAR-1 and the RNA helicase CGH-1 is required for embryonic cytokinesis in Caenorhabditis elegans
J. Cell Biol., October 24, 2005; 171(2): 267 - 279.
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 Genes Dev.Home page
T. A. Berset, E. F. Hoier, and A. Hajnal
The C. elegans homolog of the mammalian tumor suppressor Dep-1/Scc1 inhibits EGFR signaling to regulate binary cell fate decisions
Genes & Dev., June 1, 2005; 19(11): 1328 - 1340.
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 JCBHome page
H.-y. Yang, P. E. Mains, and F. J. McNally
Kinesin-1 mediates translocation of the meiotic spindle to the oocyte cortex through KCA-1, a novel cargo adapter
J. Cell Biol., May 9, 2005; 169(3): 447 - 457.
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 Mol. Biol. CellHome page
D. Poteryaev, J. M. Squirrell, J. M. Campbell, J. G. White, and A. Spang
Involvement of the Actin Cytoskeleton and Homotypic Membrane Fusion in ER Dynamics in Caenorhabditis elegans
Mol. Biol. Cell, May 1, 2005; 16(5): 2139 - 2153.
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 Mol. Cell. Biol.Home page
J. S. Martin, N. Winkelmann, M. I. R. Petalcorin, M. J. McIlwraith, and S. J. Boulton
RAD-51-Dependent and -Independent Roles of a Caenorhabditis elegans BRCA2-Related Protein during DNA Double-Strand Break Repair
Mol. Cell. Biol., April 15, 2005; 25(8): 3127 - 3139.
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 Genes Dev.Home page
V. J.P. Robert, T. Sijen, J. van Wolfswinkel, and R. H.A. Plasterk
Chromatin and RNAi factors protect the C. elegans germline against repetitive sequences
Genes & Dev., April 1, 2005; 19(7): 782 - 787.
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