Use of the piggyBac transposon for germ-line transformation of insects.

Germ-line transformation of insects is now possible with four independent transposable element vector systems. Among these, the TTAA-insertion site specific transposon, piggyBac, discovered in Trichoplusia ni, is one of the most widely used. Transformations have been achieved in a wide variety of dipterans, lepidopterans, and a coleopteran, and for many species, piggyBac transposition was first tested by plasmid-based mobility assays in cell lines and embryos. All plasmid and genomic insertions are consistent with the duplication of a TTAA insertion site, and most germ-line integrations appear to be stable, though this is largely based on stable marker phenotypes. Of the vector systems presently in use for non-drosophilids, piggyBac is the only one not currently associated with a superfamily of transposable elements, though other elements exist that share its TTAA insertion site specificity. While functional piggyBac elements have only been isolated from T. ni, nearly identical elements have been discovered in a dipteran species, Bactrocera dorsalis, and closely related elements exist in another moth species, Spodoptera frugiperda. It appears that piggyBac has recently traversed insect orders by horizontal transmission, possibly mediated by a baculovirus or other viral system. This interspecies movement has important implications for the practical use of piggyBac to create transgenic insect strains for field release.

[1]  A. Handler,et al.  Germline transformation of Drosophila melanogaster with the piggyBac transposon vector , 1999, Insect molecular biology.

[2]  T. Loukeris,et al.  Gene Transfer into the Medfly, Ceratitis capitata, with a Drosophila hydei Transposable Element , 1995, Science.

[3]  T. Miller,et al.  Precise excision and transposition of piggyBac in pink bollworm embryos , 1999, Insect molecular biology.

[4]  V. Pirrotta,et al.  A transposable P vector that confers selectable G418 resistance to Drosophila larvae , 1985, The EMBO journal.

[5]  C. Bauser,et al.  PCR analysis of insertion site specificity, transcription, and structural uniformity of the Lepidopteran transposable element IFP2 in the TN-368 cell genome , 1996, Genetica.

[6]  D. O’brochta,et al.  Interplasmid transposition of Drasopbila hobo elements in non-drosophilid insects , 1994, Molecular and General Genetics MGG.

[7]  S. Higgs,et al.  Green fluorescent protein expressed in living mosquitoes--without the requirement of transformation. , 1996, BioTechniques.

[8]  A. Handler,et al.  The lepidopteran transposon vector, piggyBac, mediates germ-line transformation in the Mediterranean fruit fly. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[9]  A. Handler,et al.  A current perspective on insect gene transformation. , 2001, Insect biochemistry and molecular biology.

[10]  A. Handler,et al.  Transformation of the Caribbean fruit fly, Anastrepha suspensa, with a piggyBac vector marked with polyubiquitin-regulated GFP. , 2001, Insect biochemistry and molecular biology.

[11]  D. O’brochta,et al.  Transposable elements and gene transformation in non-drosophilid insects. , 1996, Insect biochemistry and molecular biology.

[12]  R. L. Rabb,et al.  Movement of highly mobile insects: concepts and methodology in research. Proceedings of a conference 'Movement of selected species of Lepidoptera in the southeastern United States,' Raleigh, North Carolina. April 9-11, 1979. , 1979 .

[13]  M. Fraser,et al.  Transposon mutagenesis of baculoviruses: analysis of TFP3 lepidopteran transposon insertions at the FP locus of nuclear polyhedrosis viruses. , 1989, Gene.

[14]  Toshiki Tamura,et al.  Germline transformation of the silkworm Bombyx mori L. using a piggyBac transposon-derived vector , 2000, Nature Biotechnology.

[15]  A. Handler,et al.  Germ‐line transformation of the South American malaria vector, Anopheles albimanus, with a piggyBac/EGFP transposon vector is routine and highly efficient , 2002, Insect molecular biology.

[16]  G. Rubin,et al.  Genetic transformation of Drosophila with transposable element vectors. , 1982, Science.

[17]  Xiao-Fan Wang,et al.  Transgenic anopheline mosquitoes impaired in transmission of a malaria parasite , 2002 .

[18]  David W. Miller,et al.  A virus mutant with an insertion of a copia-like transposable element , 1982, Nature.

[19]  M. Summers,et al.  Transposon-mediated mutagenesis of a baculovirus. , 1985, Virology.

[20]  T. Hazelrigg,et al.  Implications for bcd mRNA localization from spatial distribution of exu protein in Drosophila oogenesis , 1994, Nature.

[21]  M. Summers,et al.  Acquisition of Host Cell DNA Sequences by Baculoviruses: Relationship Between Host DNA Insertions and FP Mutants of Autographa californica and Galleria mellonella Nuclear Polyhedrosis Viruses , 1983, Journal of virology.

[22]  W. Gelbart,et al.  Identification of a fully‐functional hobo transposable element and its use for germ‐line transformation of Drosophila. , 1989, The EMBO journal.

[23]  A. Handler,et al.  Mobility of P elements in drosophilids and nondrosophilids. , 1988, Proceedings of the National Academy of Sciences of the United States of America.

[24]  H. G. Wang,et al.  Assay for movement of Lepidopteran transposon IFP2 in insect cells using a baculovirus genome as a target DNA. , 1995, Virology.

[25]  J. Vlak,et al.  Horizontal Escape of the Novel Tc1-Like Lepidopteran Transposon TCp3.2 into Cydia pomonella Granulovirus , 1998, Journal of Molecular Evolution.

[26]  F. Catteruccia,et al.  piggyBac-mediated Germline Transformation of the Malaria Mosquito Anopheles stephensi Using the Red Fluorescent Protein dsRED as a Selectable Marker* , 2002, The Journal of Biological Chemistry.

[27]  A. Handler,et al.  The piggyBac transposon mediates germ‐line transformation in the Oriental fruit fly and closely related elements exist in its genome , 2000, Insect molecular biology.

[28]  M. Fraser.  The TTAA-Specific Family of Transposable Elements: Identification, Functional Characterization, and Utility for Transformation of Insects , 2000 .

[29]  I. White,et al.  Fruit Flies of Economic Significance: Their Identification and Bionomics , 1992 .

[30]  M. Scott,et al.  piggyBac‐mediated transposition in Drosophila melanogaster: an evaluation of the use of constitutive promoters to control transposase gene expression , 2001, Insect molecular biology.

[31]  S. Lukyanov,et al.  Fluorescent proteins from nonbioluminescent Anthozoa species , 1999, Nature Biotechnology.

[32]  S. Thibault,et al.  Germ‐line transformation of pink bollworm (Lepidoptera: Gelechiidae) mediated by the piggyBac transposable element , 2000, Insect molecular biology.

[33]  M. Da Rocha,et al.  3xP3-EGFP marker facilitates screening for transgenic silkworm Bombyx mori L. from the embryonic stage onwards. , 2002, Insect biochemistry and molecular biology.

[34]  A. Hua-Van,et al.  Germ line transformation of the yellow fever mosquito, Aedes aegypti, mediated by transpositional insertion of a piggyBac vector , 2002, Insect molecular biology.

[35]  N. Lobo,et al.  Transposition of the piggyBac element in embryos of Drosophila melanogaster, Aedes aegypti and Trichoplusia ni , 1999, Molecular and General Genetics MGG.

[36]  H. G. Wang,et al.  Transposon mutagenesis of baculoviruses: analysis of Trichoplusia ni transposon IFP2 insertions within the FP-locus of nuclear polyhedrosis viruses. , 1989, Virology.

[37]  A. Handler,et al.  Insect transgenesis: methods and applications. , 2000 .

[38]  C. Bauser,et al.  The minimum internal and external sequence requirements for transposition of the eukaryotic transformation vector piggyBac , 2001, Molecular Genetics and Genomics.

[39]  A. Heath,et al.  Germ‐line transformation of the Australian sheep blowfly Lucilia cuprina , 2002, Insect molecular biology.

[40]  B. Schmid,et al.  Fluorescent transformation markers for insect transgenesis. , 2002, Insect biochemistry and molecular biology.

[41]  A. Raikhel,et al.  Efficient transformation of the yellow fever mosquito Aedes aegypti using the piggyBac transposable element vector pBac[3xP3-EGFP afm]. , 2001, Insect biochemistry and molecular biology.

[42]  M. Summers,et al.  Sequence comparison of cellular and viral copies of host cell DNA insertions found in Autographa californica nuclear polyhedrosis virus. , 1990, Virology.

[43]  P. O’Farrell,et al.  A nuclear GFP that marks nuclei in living Drosophila embryos; maternal supply overcomes a delay in the appearance of zygotic fluorescence. , 1995, Developmental biology.

[44]  E. Wimmer,et al.  A versatile vector set for animal transgenesis , 2000, Development Genes and Evolution.

[45]  T. K. Stevens,et al.  Germline transformation of the malaria vector, Anopheles gambiae, with the piggyBac transposable element , 2001, Insect molecular biology.

[46]  G. Saccone,et al.  The white Gene of Ceratitis capitata: A Phenotypic Marker for Germline Transformation , 1995, Science.

[47]  M. Klingler,et al.  Genetic techniques: A universal marker for transgenic insects , 1999, Nature.

[48]  M. Chalfie,et al.  Green fluorescent protein as a marker for gene expression. , 1994, Science.

[49]  M. Benedict,et al.  The piggyBac element is capable of precise excision and transposition in cells and embryos of the mosquito, Anopheles gambiae. , 2000, Insect biochemistry and molecular biology.

[50]  J. Lis,et al.  Structure and expression of ubiquitin genes of Drosophila melanogaster , 1988, Molecular and cellular biology.

[51]  Polyubiquitin-regulated DsRed marker for transgenic insects. , 2001, BioTechniques.

[52]  A. Handler,et al.  A functional analysis of the P-element gene-transfer vector in insects. , 1993, Archives of insect biochemistry and physiology.

[53]  E. Wimmer,et al.  Genetic transformation of the housefly Musca domestica with the lepidopteran derived transposon piggyBac , 2001, Insect molecular biology.

[54]  M. Fraser,et al.  TTAA serves as the target site for TFP3 lepidopteran transposon insertions in both nuclear polyhedrosis virus and Trichoplusia ni genomes , 1993, Insect molecular biology.