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Transgenic Research

, Volume 13, Issue 5, pp 411–425 | Cite as

Formation and loss of large, unstable tandem arrays of the piggyBac transposable element in the yellow fever mosquito, Aedes aegypti

  • Zach Adelman
  • Nijole Jasinskiene
  • K. Vally
  • Corrie Peek
  • Emily Travanty
  • Ken Olson
  • Susan Brown
  • Janice Stephens
  • Dennis Knudson
  • Craig Coates
  • Anthony James
Article

Abstract

The Class II transposable element, piggyBac, was used to transform the yellow fever mosquito, Aedes aegypti. In two transformed lines only 15–30 of progeny inherited the transgene, with these individuals displaying mosaic expression of the EGFP marker gene. Southern analyses, gene amplification of genomic DNA, and plasmid rescue experiments provided evidence that these lines contained a high copy number of piggyBac transformation constructs and that much of this DNA consisted of both donor and helper plasmids. A detailed analysis of one line showed that the majority of piggyBac sequences were unit-length donor or helper plasmids arranged in a large tandem array that could be lost en masse in a single generation. Despite the presence of a transposase source and many intact donor elements, no conservative (cut and paste) transposition of piggyBac was observed in these lines. These results reveal one possible outcome of uncontrolled and/or unexpected recombination in this mosquito, and support the conclusion that further investigation is necessary before transposable elements such as piggyBac can be used as genetic drive mechanisms to move pathogen-resistance genes into mosquito populations.

rds  Genetic instability Mosquito vector PiggyBac Transgenesis 

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References

  1. Adelman ZN, Jasinskiene N and James AA (2002a) Development and applications of transgenesis in the yellow fever mosquito, Aedes aegypti. Mol Biochem Parasitol 121: 1-10.Google Scholar
  2. Adelman ZN, Sanchez-Vargas I, Travanty EA, Carlson JO, Beaty BJ, Blair CD and Olson KE (2002b) RNA silencing of dengue virus type 2 replication in transformed C6/36 mosquito cells transcribing an inverted-repeat RNA derived from the virus genome. J Virol 76: 12925-12933.Google Scholar
  3. Atkinson PW and James AA (2002) Germline transformants spreading out to many insect species. Adv Genet 47: 49-86.Google Scholar
  4. Aultman KS, Beaty BJ and Walker ED (2001) Genetically manipulated vectors of human disease: a practical overview. Trends Parasitol 17: 507-509.Google Scholar
  5. Ausubel FM, Brent R, Kingston RE, Moore DD, Seidman JG, Smith JA and Struhl K (eds.) (1987) Current protocols in molecular biology. Greene Publishing Associates and Wiley-Interscience, New York.Google Scholar
  6. Barth C, Fraser DJ and Fisher PR (1998) Co-insertional replication is responsible for tandem multimer formation during plasmid integration into the Dictyostelium genome. Plasmid 39: 141-153.Google Scholar
  7. Brown SE and Knudson DL (1997) FISH landmarks for Aedes aegypti chromosomes. Insect Mol Biol 6: 197-202.Google Scholar
  8. Brown SE, Menninger J, Difillipantonio M, Beaty BJ, Ward DC and Knudson DL (1995) Toward a physical map of Aedes aegypti. Insect Mol Biol 4: 161-167.Google Scholar
  9. Carlson CM, Dupuy AJ, Fritz S, Roberg-Perez KJ, Fletcher CF and Largaespada DA (2003) Transposon mutagenesis of the mouse germline. Genetics 165: 243-256.Google Scholar
  10. Coates CJ, Jasinskiene N, Miyashiro L and James AA (1998) Mariner transposition and transformation of the yellow fever mosquito, Aedes aegypti. Proc Natl Acad Sci USA 95: 3748-3751.Google Scholar
  11. de Lara Capurro M, Coleman J, Beerntsen BT, Myles KM, Olson KE, Rocha E, Krettli AU and James AA (2000) Virus-expressed, recombinant single-chain antibody blocks sporozoite infection of salivary glands in Plasmodium gallinaceum-infected Aedes aegypti. Am J Trop Med Hyg 62: 427-433.Google Scholar
  12. Dorer DR and Henikoff S (1994) Expansions of transgene repeats cause heterochromatin formation and gene silencing in Drosophila. Cell 77: 993-1002.Google Scholar
  13. Fraser MJ, Ciszczon T, Elick T and Bauser C (1996) Precise excision of TTAA-specific lepidopteran transposons piggyBac (IFP2) and tagalong (TFP3) from the baculovirus genome in cell lines from two species of Lepidoptera. Insect Mol Biol 5: 141-151.Google Scholar
  14. French WL, Baker RH and Kitzmiller JB (1962) Preparation of mosquito chromosomes. Mosquito News 22: 377-383.Google Scholar
  15. Gordon JW, Scangos GA, Plotkin DJ, Barbosa JA and Ruddle FH (1980) Genetic transformation of mouse embryos by microinjection of purified DNA. Proc Natl Acad Sci USA 77: 7380-7384.Google Scholar
  16. Grossman GL, Rafferty CS, Fraser MJ and Benedict MQ (2000) The piggyBac element is capable of precise excision and transposition in cells and embryos of the mosquito, Anopheles gambiae. Insect Biochem Mol Biol 30: 909-914.Google Scholar
  17. Handler AM and Harrell RA (2nd) (1999) Germline transformation of Drosophila melanogaster with the piggyBac transposon vector. Insect Mol Biol 8: 449-457.Google Scholar
  18. Handler AM and Harrell RA 2nd (2001) Transformation of the Caribbean fruit fly, Anastrepha suspensa, with a piggyBac vector marked with polyubiquitin-regulated GFP. Insect Biochem Mol Biol 31: 199-205.Google Scholar
  19. Handler AM and McCombs SD (2000) The piggyBac transposon mediates germ-line transformation in the Oriental fruit fly and closely related elements exist in its genome. Insect Mol Biol 9: 605-612.Google Scholar
  20. Handler AM, McCombs SD, Fraser MJ and Saul SH (1998) The lepidopteran transposon vector, piggyBac, mediates germ-line transformation in the Mediterranean fruit fly. Proc Natl Acad Sci USA 95: 7520-7525.Google Scholar
  21. Horn C, Jaunich B and Wimmer EA (2000) Highly sensitive, fluorescent transformation marker for Drosophila transgenesis. Dev Genes Evol 210: 623-629.Google Scholar
  22. Horn C, Offen N, Nystedt S, Hacker U and Wimmer EA (2003) piggyBac-Based Insertional Mutagenesis and Enhancer Detection as a Tool for Functional Insect Genomics. Genetics 163: 647-661.Google Scholar
  23. Horn C and Wimmer EA (2000) A versatile vector set for animal transgenesis. Dev Genes Evol 210: 630-637.Google Scholar
  24. Ito J, Ghosh A, Moreira LA, Wimmer EA and Jacobs-Lorena M (2002) Transgenic anopheline mosquitoes impaired in transmission of a malaria parasite. Nature 417: 452-455.Google Scholar
  25. Jasinskiene N, Coates CJ, Benedict MQ, Cornel AJ, Rafferty CS, James AA and Collins FH (1998) Stable transformation of the yellow fever mosquito, Aedes aegypti, with the Hermes element from the housefly. Proc Natl Acad Sci USA 95: 3743-3747.Google Scholar
  26. Kokoza V, Ahmed A, Cho WL, Jasinskiene N, James AA and Raikhel A (2000) Engineering blood meal-activated systemic immunity in the yellow fever mosquito, Aedes aegypti. Proc Natl Acad Sci USA 97: 9144-9149.Google Scholar
  27. Kokoza V, Ahmed A, Wimmer EA and Raikhel AS (2001) Efficient transformation of the yellow fever mosquito Aedes aegypti using the piggyBac transposable element vector pBac[3 · P3-EGFP afm]. Insect Biochem Mol Biol 31: 1137-1143.Google Scholar
  28. Kurenova E, Champion L, Biessmann H and Mason JM (1998) Directional gene silencing induced by a complex subtelomeric satellite from Drosophila. Chromosoma 107: 311-320.Google Scholar
  29. Li X, Lobo N, Bauser CA and Fraser MJ Jr (2001) The minimum internal and external sequence requirements for transposition of the eukaryotic transformation vector piggyBac. Mol Genet Genomics 266: 190-198.Google Scholar
  30. Lobo NF, Hua-Van A, Li X, Nolen BM and Fraser MJ Jr (2002) Germ line transformation of the yellow fever mosquito, Aedes aegypti, mediated by transpositional insertion of a piggyBac vector. Insect Mol Biol 11: 133-139.Google Scholar
  31. Lowenberger C, Charlet M, Vizioli J, Kamal S, Richman A, Christensen BM and Bulet P (1999) Antimicrobial activity spectrum, cDNA cloning, and mRNA expression of a newly isolated member of the cecropin family from the mosquito vector Aedes aegypti. J Biol Chem 274: 20092-20097.Google Scholar
  32. Monroe TJ, Muhlmann-Diaz MC, Kovach MJ, Carlson JO, Bedford JS and Beaty BJ (1992) Stable transformation of a mosquito cell line results in extraordinarily high copy numbers of the plasmid. Proc Natl Acad Sci USA 89: 5725-5729.Google Scholar
  33. Mooibroek H, Arnberg AC, de Jong B and Venema G (1985) Effect of concentration on the subsequent fate of plasmid DNA in human fibroblasts. Mol Gen Genet 199: 82-88.Google Scholar
  34. Moreira LA, Ito J, Ghosh A, Devenport M, Zieler H, Abraham EG, Crisanti A, Nolan T, Catteruccia F and Jacobs-Lorena M (2002) Bee venom phospholipase inhibits malaria parasite development in transgenic mosquitoes. J Biol Chem 277: 40839-40843.Google Scholar
  35. Nunamaker RA, Brown SE and Knudson DL (1999) Fluorescence in situ hybridization landmarks for chromosomes for Culicoides variipennis (Diptera: Ceratopogonidae). J Med Entomol 36: 171-175.Google Scholar
  36. O'Brochta DA, Sethuraman N, Wilson R, Hice RH, Pinkerton AC, Levesque CS, Bideshi DK, Jasinskiene N, Coates CJ, James AA, Lehane MJ and Atkinson PW (2003) Gene vector and transposable element behavior in mosquitoes. J Exp Biol 206: 3823-3834.Google Scholar
  37. Pawlowski WP and Somers DA (1998) Transgenic DNA integrated into the oat genome is frequently interspersed by host DNA. Proc Natl Acad Sci USA 95: 12106-12110.Google Scholar
  38. Perera OP, Harrell IR and Handler AM (2002) Germ-line transformation of the South American malaria vector, Anopheles albimanus, with a piggyBac/EGFP transposon vector is routine and highly efficient. Insect Mol Biol 11: 291-297.Google Scholar
  39. Perucho M, Hanahan D and Wigler M (1980) Genetic and physical linkage of exogenous sequences in transformed cells. Cell 22: 309-317.Google Scholar
  40. Plessis A and Dujon B (1993) Multiple tandem integrations of transforming DNA sequences in yeast chromosomes suggest a mechanism for integrative transformation by homologous recombination. Gene 134: 41-50.Google Scholar
  41. Pomerantz BJ, Naujokas M and Hassell JA (1983) Homologous recombination between transfected DNAs. Mol Cell Biol 3: 1680-1685.Google Scholar
  42. Ribeiro JM and Kidwell MG (1994) Transposable elements as population drive mechanisms: specification of critical parameter values. J Med Entomol 31: 10-16.Google Scholar
  43. Robins DM, Ripley S, Henderson AS and Axel R (1981) Transforming DNA integrates into the host chromosome. Cell 23: 29-39.Google Scholar
  44. Roche SE and Rio DC (1998) Trans-silencing by P elements inserted in subtelomeric heterochromatin involves the Drosophila Polycomb group gene, Enhancer of zeste. Genetics 149: 1839-1855.Google Scholar
  45. Ronsseray S, Boivin A and Anxolabehere D (2001) P-Element repression in Drosophila melanogaster by variegating clusters of P-lacZ-white transgenes Genetics 159: 1631-1642.Google Scholar
  46. Scangos G and Ruddle FH (1981) Mechanisms and applications of DNA-mediated gene transfer in mammalian cells-a review. Gene 14: 1-10.Google Scholar
  47. Smartt CT, Kiley LM, Hillyer JF, Dasgupta R and Christensen BM (2001) Aedes aegypti glutamine synthetase: expression and gene structure. Gene 274: 35-45.Google Scholar
  48. Uhlirova M, Asahina M, Riddiford LM and Jindra M (2002) Heat-inducible transgenic expression in the silkmoth Bombyx mori. Dev Genes Evol 212: 145-151.Google Scholar
  49. Walter MF, Bozorgnia L, Maheshwari A and Biessmann H (2001) The rate of terminal nucleotide loss from a telomere of the mosquito Anopheles gambiae. Insect Mol Biol 10: 105-110.Google Scholar
  50. Wilson R, Orsetti J, Klocko AD, Aluvihare C, Peckham E, Atkinson PW, Lehane MJ and O'Brochta DA (2003) Postintegration behavior of a Mos1 mariner gene vector in Aedes aegypti. Insect Biochem Mol Biol 33: 853-863.Google Scholar
  51. Wong EA and Capecchi MR (1987) Homologous recombination between coinjected DNA sequences peaks in early to mid-S phase. Mol Cell Biol 7: 2294-2295.Google Scholar

Copyright information

© Kluwer Academic Publishers 2004

Authors and Affiliations

  • Zach Adelman
    • 1
  • Nijole Jasinskiene
    • 1
  • K. Vally
    • 2
  • Corrie Peek
    • 1
  • Emily Travanty
    • 3
  • Ken Olson
    • 3
  • Susan Brown
    • 4
  • Janice Stephens
    • 4
  • Dennis Knudson
    • 4
  • Craig Coates
    • 2
  • Anthony James
    • 1
  1. 1.Department of Molecular Biology and BiochemistryUniversity of CaliforniaUSA
  2. 2.Department of EntomologyCollege of Agriculture and Life Sciences, Texas A&M UniversityUSA
  3. 3.Department of MicrobiologyImmunology and PathologyUSA
  4. 4.Department of Bioagricultural Sciences and Pest ManagementCollege of Agricultural SciencesUSA

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