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PMID: 19754740 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Homing endonucleases catalyze double-stranded DNA breaks and somatic transgene excision in Aedes aegypti.

Insect molecular biology ·Vol. 18 ·No. 5 ·2009-10-00 ·Pages 623-33

Traver BE, Anderson MA, Adelman ZN

Abstract

Aedes aegypti is a major vector of arthropod-borne viruses such as yellow fever virus and dengue viruses. Efforts to discern the function of genes involved in important behaviours, such as vector competence and host seeking through reverse genetics, would greatly benefit from the ability to generate targeted gene disruptions. Homing endonucleases are selfish elements which catalyze double-stranded DNA (dsDNA) breaks in a sequence-specific manner. In this report we demonstrate that the homing endonucleases I-PpoI, I-SceI, I-CreI and I-AniI are all able to induce dsDNA breaks in adult female Ae. aegypti chromosomes as well as catalyze the somatic excision of a transgene. These experiments provide evidence that homing endonucleases can be used to manipulate the genome of this important disease vector.

MeSH Terms
Aedes/enzymology,genetics,virology Animals Base Sequence Biocatalysis Biological Assay Blotting, Southern DNA Breaks, Double-Stranded DNA Footprinting DNA Repair DNA, Recombinant/genetics Deoxyribonucleases, Type II Site-Specific/metabolism Molecular Sequence Data Plasmids/genetics Protein Transport Sindbis Virus/physiology Transgenes/genetics
Chemicals
DNA, Recombinant Deoxyribonucleases, Type II Site-Specific
Authors & Affiliations
3 authors, click to expand affiliations / ORCID
Traver B E
Department of Entomology, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061, USA.
Anderson M A E
Adelman Z N
References (49)
49 references, click to expand
  1. Homing endonuclease mediated gene targeting in Anopheles gambiae cells and embryos.
    Nucleic Acids Res. 2007;35(17):5922-33 PMID: 17726053
  2. Repair of ionizing radiation-induced DNA double-strand breaks by non-homologous end-joining.
    Biochem J. 2009 Feb 1;417(3):639-50 PMID: 19133841
  3. A combinatorial approach to create artificial homing endonucleases cleaving chosen sequences.
    Nucleic Acids Res. 2006;34(22):e149 PMID: 17130168
  4. Efficient transformation of the yellow fever mosquito Aedes aegypti using the piggyBac transposable element vector pBac[3xP3-EGFP afm].
    Insect Biochem Mol Biol. 2001 Nov 1;31(12):1137-43 PMID: 11583926
  5. Stable transformation of the yellow fever mosquito, Aedes aegypti, with the Hermes element from the housefly.
    Proc Natl Acad Sci U S A. 1998 Mar 31;95(7):3743-7 PMID: 9520437
  6. Ends-out, or replacement, gene targeting in Drosophila.
    Proc Natl Acad Sci U S A. 2003 Mar 4;100(5):2556-61 PMID: 12589026
  7. Impact of the KU80 pathway on NHEJ-induced genome rearrangements in mammalian cells.
    Mol Cell. 2004 Jun 4;14(5):611-23 PMID: 15175156
  8. Rapid "open-source" engineering of customized zinc-finger nucleases for highly efficient gene modification.
    Mol Cell. 2008 Jul 25;31(2):294-301 PMID: 18657511
  9. Effects of an opal termination codon preceding the nsP4 gene sequence in the O'Nyong-Nyong virus genome on Anopheles gambiae infectivity.
    J Virol. 2006 May;80(10):4992-7 PMID: 16641290
  10. Infectious Sindbis virus transient expression vectors for studying antigen processing and presentation.
    Proc Natl Acad Sci U S A. 1992 Apr 1;89(7):2679-83 PMID: 1372987
  11. Optimization of in vivo activity of a bifunctional homing endonuclease and maturase reverses evolutionary degradation.
    Nucleic Acids Res. 2009 Feb;37(3):877-90 PMID: 19103658
  12. Mariner transposition and transformation of the yellow fever mosquito, Aedes aegypti.
    Proc Natl Acad Sci U S A. 1998 Mar 31;95(7):3748-51 PMID: 9520438
  13. Conservative homologous recombination preferentially repairs DNA double-strand breaks in the S phase of the cell cycle in human cells.
    Nucleic Acids Res. 2004 Jul 13;32(12):3683-8 PMID: 15252152
  14. A targeted gene knockout in Drosophila.
    Genetics. 2001 Mar;157(3):1307-12 PMID: 11238415
  15. I-PpoI and I-CreI homing site sequence degeneracy determined by random mutagenesis and sequential in vitro enrichment.
    J Mol Biol. 1998 Jul 17;280(3):345-53 PMID: 9665841
  16. I-SceI meganuclease mediates highly efficient transgenesis in fish.
    Mech Dev. 2002 Oct;118(1-2):91-8 PMID: 12351173
  17. Highly efficient sex chromosome interchanges produced by I-CreI expression in Drosophila.
    Genetics. 2005 Nov;171(3):1103-14 PMID: 16020774
  18. Genome sequence of Aedes aegypti, a major arbovirus vector.
    Science. 2007 Jun 22;316(5832):1718-23 PMID: 17510324
  19. A transgenic sensor strain for monitoring the RNAi pathway in the yellow fever mosquito, Aedes aegypti.
    Insect Biochem Mol Biol. 2008 Jul;38(7):705-13 PMID: 18549956
  20. High efficiency site-specific genetic engineering of the mosquito genome.
    Insect Mol Biol. 2006 Apr;15(2):129-36 PMID: 16640723
  21. Targeting the X chromosome during spermatogenesis induces Y chromosome transmission ratio distortion and early dominant embryo lethality in Anopheles gambiae.
    PLoS Genet. 2008 Dec;4(12):e1000291 PMID: 19057670
  22. Dengue.
    Lancet. 2007 Nov 10;370(9599):1644-52 PMID: 17993365
  23. Gene targeting by homologous recombination in Drosophila.
    Science. 2000 Jun 16;288(5473):2013-8 PMID: 10856208
  24. Mutations altering the cleavage specificity of a homing endonuclease.
    Nucleic Acids Res. 2002 Sep 1;30(17):3870-9 PMID: 12202772
  25. Gene drive systems for insect disease vectors.
    Nat Rev Genet. 2006 Jun;7(6):427-35 PMID: 16682981
  26. Engineering of large numbers of highly specific homing endonucleases that induce recombination on novel DNA targets.
    J Mol Biol. 2006 Jan 20;355(3):443-58 PMID: 16310802
  27. The expression of chloramphenicol acetyltransferase in Aedes albopictus (C6/36) cells and Aedes triseriatus mosquitoes using a double subgenomic recombinant Sindbis virus.
    Insect Biochem Mol Biol. 1994 Jan;24(1):39-48 PMID: 8111422
  28. Generation of highly site-specific DNA double-strand breaks in human cells by the homing endonucleases I-PpoI and I-CreI.
    Biochem Biophys Res Commun. 1999 Feb 5;255(1):88-93 PMID: 10082660
  29. Homing endonuclease I-CreI derivatives with novel DNA target specificities.
    Nucleic Acids Res. 2006;34(17):4791-800 PMID: 16971456
  30. Site-specific selfish genes as tools for the control and genetic engineering of natural populations.
    Proc Biol Sci. 2003 May 7;270(1518):921-8 PMID: 12803906
  31. The population genetics of using homing endonuclease genes in vector and pest management.
    Genetics. 2008 Aug;179(4):2013-26 PMID: 18660532
  32. An efficient method to generate chromosomal rearrangements by targeted DNA double-strand breaks in Drosophila melanogaster.
    Genome Res. 2004 Jul;14(7):1382-93 PMID: 15197166
  33. Directed evolution and substrate specificity profile of homing endonuclease I-SceI.
    J Am Chem Soc. 2006 Feb 22;128(7):2477-84 PMID: 16478204
  34. Homing endonucleases: keeping the house in order.
    Nucleic Acids Res. 1997 Sep 1;25(17):3379-88 PMID: 9254693
  35. The 5' regulatory region from the Drosophila pseudoobscura hsp82 gene results in a high level of reporter gene expression in Lucilia cuprina embryos.
    Gene. 1996 Oct 10;175(1-2):199-201 PMID: 8917099
  36. Altered target site specificity variants of the I-PpoI His-Cys box homing endonuclease.
    Nucleic Acids Res. 2007;35(17):5839-50 PMID: 17720708
  37. High efficiency, site-specific excision of a marker gene by the phage P1 cre-loxP system in the yellow fever mosquito, Aedes aegypti.
    Nucleic Acids Res. 2003 Nov 15;31(22):e147 PMID: 14602940
  38. Intraspecific DNA variation in nuclear genes of the mosquito Aedes aegypti.
    Insect Mol Biol. 2003 Dec;12(6):631-9 PMID: 14986924
  39. In vivo selection of engineered homing endonucleases using double-strand break induced homologous recombination.
    Nucleic Acids Res. 2005 Nov 23;33(20):e178 PMID: 16306233
  40. Development of an orally infectious Sindbis virus transducing system that efficiently disseminates and expresses green fluorescent protein in Aedes aegypti.
    Insect Mol Biol. 2003 Apr;12(2):107-16 PMID: 12653932
  41. Purification, cloning, and characterization of the CEL I nuclease.
    Biochemistry. 2000 Apr 4;39(13):3533-41 PMID: 10736152
  42. Green fluorescent protein expressed in living mosquitoes--without the requirement of transformation.
    Biotechniques. 1996 Oct;21(4):660-4 PMID: 8891217
  43. The role of double-strand break-induced allelic homologous recombination in somatic plant cells.
    Plant J. 2002 Nov;32(3):277-84 PMID: 12410807
  44. Genetic manipulation of genomes with rare-cutting endonucleases.
    Trends Genet. 1996 Jun;12(6):224-8 PMID: 8928227
  45. I-SceI endonuclease, a new tool for studying DNA double-strand break repair mechanisms in Drosophila.
    Genetics. 1999 Jul;152(3):1037-44 PMID: 10388822
  46. Characterization of homing endonucleases.
    Methods. 2002 Nov;28(3):365-73 PMID: 12431440
  47. Engineered I-CreI derivatives cleaving sequences from the human XPC gene can induce highly efficient gene correction in mammalian cells.
    J Mol Biol. 2007 Aug 3;371(1):49-65 PMID: 17561112
  48. Targeted gene knockout in mammalian cells by using engineered zinc-finger nucleases.
    Proc Natl Acad Sci U S A. 2008 Apr 15;105(15):5809-14 PMID: 18359850
  49. The homologous chromosome is an effective template for the repair of mitotic DNA double-strand breaks in Drosophila.
    Genetics. 2003 Dec;165(4):1831-42 PMID: 14704169
Article Info
Journal
Insect molecular biology
Abbr.
Insect Mol Biol
ISSN
1365-2583
DOI
10.1111/j.1365-2583.2009.00905.x
Published
2009-10-00
Pages
623-33
Language
English
Region
England
NLM ID
9303579
PMCID
PMC3606018
Subset
IM
Grants
NIAID NIH HHS · R21 AI071208 · United States
NIAID NIH HHS · 1R21AI071208-01A1 · United States
External Links
PubMed source DOI: 10.1111/j.1365-2583.2009.00905.x PMC Full Text View journal details
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