Skip to main content

Advertisement

Log in

Site-directed integration of transgenes: transposons revisited using DNA-binding-domain technologies

  • Published:
Genetica Aims and scope Submit manuscript

Abstract

In the last 20 years, tools derived from DNA transposons have made major contributions to genetic studies from gene delivery to gene discovery. Various complementary and fairly ubiquitous DNA vehicles have been developed. Although many transposons are efficient DNA vehicles, they appear to have limited ability to target specific sequences, since all that is required at the integration locus is the presence of a short 2- to 4-bp sequence. Consequently, insertions mediated by transposon-based vectors occur somewhat randomly. In the past 5 years, strategies have emerged to enhance the site-specificity of transposon-based vectors, and to avoid random integrations. The first proposes that new target site specificity could be grafted onto a transposase by adding a new DNA-binding domain. Alternative strategies consist of indirectly targeting either the transposase or the transposon to a chosen genomic locus. The most important information available about each strategy are presented, and limitations and future prospects are discussed.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Akerley BJ, Rubin EJ, Camilli A, Lampe DJ, Robertson HM, Mekalanos JJ (1998) Systematic identification of essential genes by in vitro mariner mutagenesis. Proc Natl Acad Sci USA 95:8927–8932

    Article  CAS  PubMed  Google Scholar 

  • Augé-Gouillou C, Hamelin MH, Demattéi MV, Périquet M, Bigot Y (2001) The ITR binding domain of the Mariner MOS1 transposase. Mol Genet Genomics 265:58–65

    Article  PubMed  Google Scholar 

  • Augé-Gouillou C, Brillet B, Germon S, Hamelin MH, Bigot Y (2005) Mariner Mos1 transposase dimerizes prior to ITR binding. J Mol Biol 351:117–130

    Article  PubMed  Google Scholar 

  • Aye M, Irwin B, Beliakova-Bethell N, Chen E, Garrus J, Sandmeyer S (2004) Host factors that affect Ty3 retrotransposition in Saccharomyces cerevisiae. Genetics 168:1159–1176

    Article  CAS  PubMed  Google Scholar 

  • Bessereau JL, Wright A, Williams DC, Schuske K, Davis MW, Jorgensen EM (2001) Mobilization of a Drosophila transposon in the Caenorhabditis elegans germ line. Nature 413:70–74

    Article  CAS  PubMed  Google Scholar 

  • Bushman FD (2003) Targeting survival: integration site selection by retroviruses and LTR-retrotransposons. Cell 115:135–138

    Article  CAS  PubMed  Google Scholar 

  • Cadiñanos J, Bradley A (2007) Generation of an inducible and optimized piggyBac transposon system. Nucleic Acids Res 35(12):e87

    Article  PubMed  Google Scholar 

  • Carapuça E, Azzoni AR, Prazeres DM, Monteiro GA, Mergulhão FJ (2007) Time-course determination of plasmid content in eukaryotic and prokaryotic cells using real-time PCR. Mol Biotechnol 37:120–126

    Article  PubMed  Google Scholar 

  • Coates CJ, Jasinskiene N, Miyashiro L, James AA (1998) Mariner transposition and transformation of the yellow fever mosquito, Aedes aegypti. Proc Natl Acad Sci USA 95:3748–3751

    Article  CAS  PubMed  Google Scholar 

  • Conrath KE, Lauwereys M, Wyns L, Muyldermans S (2001) Camel single-domains antibodies as modular building units for bispecific and bivalent antibody constructs. J Biol Chem 276:7346–7350

    Article  CAS  Google Scholar 

  • Crénès G, Ivo D, Hérisson J, Dion S, Renault S, Bigot Y, Petit A (2009) The bacterial Tn9 chloramphenicol resistance gene: an attractive DNA segment for Mos1 mariner insertions. Mol Genet Genomics 281:315–328

    Article  PubMed  Google Scholar 

  • Fadool JM, Hartl DL, Dowling JE (1998) Transposition of the mariner element from Drosophila mauritiana in zebrafish. Proc Natl Acad Sci USA 95:5182–5186

    Article  CAS  PubMed  Google Scholar 

  • Granger L, Martin E, Ségalat L (2004) Mos as a tool for genome-wide insertional mutagenesis in Caenorhabditis elegans: results of a pilot study. Nucleic Acids Res 32:e117

    Article  PubMed  Google Scholar 

  • Guddat LW, Herron JN, Edmundson AB (1993) Three-dimensional structure of a human immunoglobulin with a hinge deletion. Proc Natl Acad Sci USA 90:4271–4275

    Article  CAS  PubMed  Google Scholar 

  • Gueiros-Filho FJ, Beverley SM (1997) Trans-kingdom transposition of the Drosophila element mariner within the protozoan Leishmania. Science 276:1716–1719

    Article  CAS  PubMed  Google Scholar 

  • Guimond N, Bideshi DK, Pinkerton AC, Atkinson PW, O’Brochta DA (2003) Patterns of Hermes transposition in Drosophila melanogaster. Mol Genet Genomics 268:779–790

    CAS  PubMed  Google Scholar 

  • Hama C, Ali Z, Kornberg TB (1990) Region-specific recombination and expression are directed by portions of the Drosophila engrailed promoter. Genes Dev 4:1079–1093

    Article  CAS  PubMed  Google Scholar 

  • Handler AM (2002) Prospects for using genetic transformation for improved SIT and new biocontrol methods. Genetica 116:137–149

    Article  CAS  PubMed  Google Scholar 

  • Handler AM, Harrell RAII (1998) Germline transformation of Drosophila melanogaster with the piggyBac transposon vector. Insect Mol Biol 8:449–457

    Article  Google Scholar 

  • Ivics Z, Hackett PB, Plasterk RH, Izsvák Z (1997) Molecular reconstruction of Sleeping Beauty, a Tc1-like transposon from fish, and its transposition in human cells. Cell 14:501–510

    Article  Google Scholar 

  • Ivics Z, Katzer A, Stüwe EE, Fiedler D, Knespel S, Izsvák Z (2007) Targeted Sleeping Beauty transposition in human cells. Mol Ther 15:1137–1144

    CAS  PubMed  Google Scholar 

  • Izsvák Z, Khare D, Behlke J, Heinemann U, Plasterk RH, Ivics Z (2002) Involvement of a bifunctional, paired-like DNA-binding domain and a transpositional enhancer in Sleeping Beauty transposition. J Biol Chem 277:34581–34588

    Article  PubMed  Google Scholar 

  • Kassis JA, Noll E, VanSickle EP, Odenwald WF, Perrimon N (1992) Altering the insertional specificity of a Drosophila transposable element. Proc Natl Acad Sci USA 89:1919–1923

    Article  CAS  PubMed  Google Scholar 

  • Kavoosi M, Creagh AL, Kilburn DG, Haynes CA (2007) Strategy for selecting and characterizing linker peptides for CBM9-tagged fusion proteins expressed in Escherichia coli. Biotechnol Bioeng 98:599–610

    Article  CAS  PubMed  Google Scholar 

  • Keng VW, Yae K, Hayakawa T, Mizuno S, Uno Y, Yusa K, Kokubu C, Kinoshita T, Akagi K, Jenkins NA, Copeland NG, Horie K, Takeda J (2005) Region-specific saturation germline mutagenesis in mice using the Sleeping Beauty transposon system. Nat Methods 2:763–769

    Article  CAS  PubMed  Google Scholar 

  • Keravala A, Liu D, Lechman ER, Wolfe D, Nash JA, Lampe DJ, Robbins PD (2006) Hyperactive Himar1 transposase mediates transposition in cell culture and enhances gene expression in vivo. Hum Gene Ther 17:1006–1018

    Article  CAS  PubMed  Google Scholar 

  • Lampe DJ, Churchill ME, Robertson HM (1996) A purified mariner transposase is sufficient to mediate transposition in vitro. EMBO J 15(5):470–479

    Google Scholar 

  • Lipkow K, Buisine N, Chalmers R (2004) Promiscuous target interactions in the mariner transposon Himar1. J Biol Chem 279:48569–48575

    Article  CAS  PubMed  Google Scholar 

  • Liu X, Liu M, Xue Z, Pan Q, Wu L, Long Z, Xia K, Liang D, Xia J (2007) Non-viral ex vivo transduction of human hepatocyte cells to express factor VIII using a human ribosomal DNA-targeting vector. J Thromb Haemost 5:347–351

    Article  CAS  PubMed  Google Scholar 

  • Luo G, Ivics Z, Izsvák Z, Bradley A (1998) Chromosomal transposition of a Tc1/mariner-like element in mouse embryonic stem cells. Proc Natl Acad Sci USA 95:10769–10773

    Article  CAS  PubMed  Google Scholar 

  • Maragathavally KJ, Kaminski JM, Coates CJ (2006) Chimeric Mos1 and piggyBac transposases result in site-directed integration. FASEB J 20:1880–1882

    Article  CAS  PubMed  Google Scholar 

  • McNamara AR, Ford KG (2000) A novel four zinc-finger protein targeted against p190(BcrAbl) fusion oncogene cDNA: utilisation of zinc-finger recognition codes. Nucleic Acids Res 28:4865–4872

    Article  CAS  PubMed  Google Scholar 

  • Medhora M, Maruyama K, Hartl DL (1991) Molecular and functional analysis of the mariner mutator element Mos1 in Drosophila. Genetics 128:311–318

    CAS  PubMed  Google Scholar 

  • Palazzoli F, Carnus E, Wells DJ, Bigot Y (2008) Sustained transgene expression using non-viral enzymatic systems for stable chromosomal integration. Curr Gene Ther 8:367–390

    Article  CAS  PubMed  Google Scholar 

  • Porteus MH, Carroll D (2005) Gene targeting using zinc finger nucleases. Nat Biotechnol 23:967–973

    Article  CAS  PubMed  Google Scholar 

  • Rubin GM, Spradling AC (1982) Genetic transformation of Drosophila with transposable element vectors. Science 218:348–353

    Article  CAS  PubMed  Google Scholar 

  • Schweidenback CT, Baker TA (2008) Dissecting the roles of MuB in Mu transposition: ATP regulation of DNA binding is not essential for target delivery. Proc Natl Acad Sci USA 105:12101–12107

    Article  CAS  PubMed  Google Scholar 

  • Sharma N, Moldt B, Dalsgaard T, Jensen TG, Mikkelsen JG (2008) Regulated gene insertion by steroid-induced PhiC31 integrase. Nucleic Acids Res 36(11):e67

    Article  PubMed  Google Scholar 

  • Sherman A, Dawson A, Mather C, Gilhooley H, Li Y, Mitchell R, Finnegan D, Sang H (1998) Transposition of the Drosophila element mariner into the chicken germ line. Nat Biotechnol 16:1050–1053

    Article  CAS  PubMed  Google Scholar 

  • Sumitani M, Yamamoto DS, Oishi K, Lee JM, Hatakeyama M (2003) Germline transformation of the sawfly, Athalia rosae (Hymenoptera: Symphyta), mediated by a piggyBac-derived vector. Insect Biochem Mol Biol 33:449–458

    Article  CAS  PubMed  Google Scholar 

  • Szabó M, Müller F, Kiss J, Balduf C, Strähle U, Olasz F (2003) Transposition and targeting of the prokaryotic mobile element IS30 in zebrafish. FEBS Lett 550:46–50

    Article  PubMed  Google Scholar 

  • Takeda J, Keng VW, Horie K (2007) Germline mutagenesis mediated by Sleeping Beauty transposon system in mice. Genome Biol 8(Suppl 1):S14

    Article  PubMed  Google Scholar 

  • Tan W, Zhu K, Segal DJ, Barbas CF 3rd, Chow SA (2004) Fusion proteins consisting of human immunodeficiency virus type 1 integrase and the designed polydactyl zinc finger protein E2C direct integration of viral DNA into specific sites. J Virol 78:1301–1313

    Article  CAS  PubMed  Google Scholar 

  • Tan W, Dong Z, Wilkinson TA, Barbas CF 3rd, Chow SA (2006) Human immunodeficiency virus type 1 incorporated with fusion proteins consisting of integrase and the designed polydactyl zinc finger protein E2C can bias integration of viral DNA into a predetermined chromosomal region in human cells. J Virol 80:1939–1948

    Article  CAS  PubMed  Google Scholar 

  • van der Linden R, de Geus B, Stok W, Bos W, van Wassenaar D, Verrips T, Frenken L (2000) Induction of immune responses and molecular cloning of the heavy chain antibody repertoire of Lama glama. J Immunol Methods 240:185–195

    Article  PubMed  Google Scholar 

  • Vigdal TJ, Kaufman CD, Izsvák Z, Voytas DF, Ivics Z (2002) Common physical properties of DNA affecting target site selection of Sleeping beauty and other Tc1/mariner transposable elements. J Mol Biol 323:441–452

    Article  CAS  PubMed  Google Scholar 

  • Voigt K, Ivics Z (2008) Targeted of SB transposon by a fusion of the SB Tpase with zinc fingers. International congress on Transposable Elements, St Malo

    Google Scholar 

  • Walisko O, Schorn A, Rolfs F, Devaraj A, Miskey C, Izsvák Z, Ivics Z (2006) Transcriptional activities of the Sleeping Beauty transposon and shielding its genetic cargo with insulators. Mol Ther 16:359–369

    Article  Google Scholar 

  • Wang W, Lin C, Lu D, Ning Z, Cox T, Melvin D, Wang X, Bradley A, Liu P (2008) Chromosomal transposition of PiggyBac in mouse embryonic stem cells. Proc Natl Acad Sci USA 105:9290–9295

    Article  CAS  PubMed  Google Scholar 

  • Wilson MH, Kaminski JM, George AL Jr (2005) Functional zinc finger/sleeping beauty transposase chimeras exhibit attenuated overproduction inhibition. FEBS Lett 579:6205–6209

    Article  CAS  PubMed  Google Scholar 

  • Wu SC, Meir YJ, Coates CJ, Handler AM, Pelczar P, Moisyadi S, Kaminski JM (2006) piggyBac is a flexible and highly active transposon as compared to Sleeping beauty, Tol2, and Mos1 in mammalian cells. Proc Natl Acad Sci USA 103:15008–15013

    Article  CAS  PubMed  Google Scholar 

  • Yant SR, Huang Y, Akache B, Kay MA (2007) Site-directed transposon integration in human cells. Nucleic Acids Res 35:1–13

    Article  Google Scholar 

  • Zhu Y, Dai J, Fuerst PG, Voytas DF (2003) Controlling integration specificity of a yeast retrotransposon. Proc Natl Acad Sci USA 100:5891–5895

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

This work was supported by the University of François Rabelais of Tours, the C.N.R.S., the French Ministère de l’Education Nationale, de la Recherche et de la Technologie (MENRT) and funded by grants from the European Commission (Project SyntheGeneDelivery, No. 018716), and the Agence Nationale de la Recherche (ANR POGM 05-003). The English text has been revised by Dr M. Ghosh.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Sylvaine Renault.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Demattei, MV., Thomas, X., Carnus, E. et al. Site-directed integration of transgenes: transposons revisited using DNA-binding-domain technologies. Genetica 138, 531–540 (2010). https://doi.org/10.1007/s10709-009-9390-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10709-009-9390-y

Keywords

Navigation