Skip to main content
SpringerLink
Log in

Sperm-Mediated Genetic Modifications

  • Protocol
  • First Online:
Transgenic Mouse

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2066))

  • 2255 Accesses

Abstract

The ability to introduce controlled modifications of the genome of animals represents an important tool for biomedical and veterinary research. Among transgenic techniques, we describe here the sperm-mediated gene transfer method that is based on the spontaneous ability of sperm cells to bind and internalize exogenous DNA and to carry it to the oocyte during fertilization, producing genetically modified animals.

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

Access this chapter

Log in via an institution
Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 109.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 219.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Gordon JW, Scangos GA, Plotkin DJ et al (1980) Genetic transformation of mouse embryos by microinjection of purified DNA. Proc Natl Acad Sci U S A 77:7380–7384

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Tan W, Proudfoot C, Lillico SG, Whitelaw CBA (2016) Gene targeting, genome editing: from Dolly to editors. Transgenic Res 25:273–287. https://doi.org/10.1007/s11248-016-9932-x

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Vandamme TF (2015) Rodent models for human diseases. Eur J Pharmacol 759:84–89. https://doi.org/10.1016/j.ejphar.2015.03.046

    Article  CAS  PubMed  Google Scholar 

  4. Th Rülicke, X Montagutelli, B Pintado, R Thon, H J Hedrich, (2016) FELASA guidelines for the production and nomenclature of transgenic rodents. Laboratory Animals 41(3):301–311

    Google Scholar 

  5. Gordon JW, Ruddle FH (1981) Integration and stable germ line transmission of genes injected into mouse pronuclei. Science 214:1244–1246

    Article  CAS  PubMed  Google Scholar 

  6. Belizário JE, Akamini P, Wolf P et al (2012) New routes for transgenesis of the mouse. J Appl Genet 53:295–315. https://doi.org/10.1007/s13353-012-0096-y

    Article  CAS  PubMed  Google Scholar 

  7. Smithies O, Gregg RG, Boggs SS et al (1985) Insertion of DNA sequences into the human chromosomal beta-globin locus by homologous recombination. Nature 317:230–234

    Article  CAS  PubMed  Google Scholar 

  8. Capecchi MR (1989) Altering the genome by homologous recombination. Science 244:1288–1292. https://doi.org/10.1126/science.2660260

    Article  CAS  PubMed  Google Scholar 

  9. Brackett BG, Baranska W, Sawicki W, Koprowski H (1971) Uptake of heterologous genome by mammalian spermatozoa and its transfer to ova through fertilization. Proc Natl Acad Sci U S A 68:353–357

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  10. Lavitrano M, Camaioni A, Fazio VM et al (1989) Sperm cells as vectors for introducing foreign DNA into eggs: genetic transformation of mice. Cell 57:717–723

    Article  CAS  PubMed  Google Scholar 

  11. Francolini M, Lavitrano M, Lamia C et al (1993) Evidence for nuclear internalization of exogenous DNA into mammalian sperm cells. Mol Reprod Dev 34:133–139

    Article  CAS  PubMed  Google Scholar 

  12. Zani M, Lavitrano M, French D et al (1995) The mechanism of binding of exogenous DNA to sperm cells: factors controlling the DNA uptake. Exp Cell Res 217:57–64. https://doi.org/10.1006/excr.1995.1063

    Article  CAS  PubMed  Google Scholar 

  13. Lavitrano M, Maione B, Forte E et al (1997) The interaction of sperm cells with exogenous DNA: a role of CD4 and major histocompatibility complex class II molecules. Exp Cell Res 233(1):58–62. https://doi.org/10.1006/excr.1997.3534

    Article  Google Scholar 

  14. Lavitrano M, Busnelli M, Cerrito MG et al (2006) Sperm-mediated gene transfer. Reprod Fertil Dev 18:19–23

    Article  CAS  PubMed  Google Scholar 

  15. Magnano A, Giordano R, Moscufo N et al (1998) Sperm/DNA interaction: integration of foreign DNA sequences in the mouse sperm genome. J Reprod Immunol 41:187–196

    Article  CAS  PubMed  Google Scholar 

  16. Lavitrano M, Bacci ML, Forni M et al (2002) Efficient production by sperm-mediated gene transfer of human decay accelerating factor (hDAF) transgenic pigs for xenotransplantation. Proc Natl Acad Sci U S A 99:14230–14235. https://doi.org/10.1073/pnas.222550299

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Maione B, Pittoggi C, Achene L et al (1997) Activation of endogenous nucleases in mature sperm cells upon interaction with exogenous DNA. DNA Cell Biol 16:1087–1097. https://doi.org/10.1089/dna.1997.16.1087

    Article  CAS  PubMed  Google Scholar 

  18. Lavitrano M, Giovannoni R, Cerrito MG (2013) Methods for sperm-mediated gene transfer. Methods Mol Biol 927:519–529. https://doi.org/10.1007/978-1-62703-038-0_44

    Article  CAS  PubMed  Google Scholar 

  19. Lavitrano M, Forni M, Bacci ML et al (2003) Sperm mediated gene transfer in pig: selection of donor boars and optimization of DNA uptake. Mol Reprod Dev 64:284–291. https://doi.org/10.1002/mrd.10230

    Article  CAS  PubMed  Google Scholar 

  20. Cong L, Cao G, Renyu X et al (2011) Reducing blood glucose level in TIDM mice by orally administering the silk glands of transgenic hIGF-I silkworms. Biochem Biophys Res Commun 410:721–725. https://doi.org/10.1016/j.bbrc.2011.05.157

    Article  CAS  PubMed  Google Scholar 

  21. Sin FY, Walker SP, Symonds JE et al (2000) Electroporation of salmon sperm for gene transfer: efficiency, reliability, and fate of transgene. Mol Reprod Dev 56:285–288. https://doi.org/10.1002/(SICI)1098-2795(200006)56:2+<285::AID-MRD16>3.0.CO;2-4

    Article  CAS  PubMed  Google Scholar 

  22. Campos VF, Amaral MG, Seixas FK et al (2011) Exogenous DNA uptake by South American catfish (Rhamdia quelen) spermatozoa after seminal plasma removal. Anim Reprod Sci 126:136–141. https://doi.org/10.1016/j.anireprosci.2011.05.004

    Article  CAS  PubMed  Google Scholar 

  23. Collares T, Campos VF, De Leon PM et al (2011) Transgene transmission in chickens by sperm-mediated gene transfer after seminal plasma removal and exogenous DNA treated with dimethylsulfoxide or N,N-dimethylacetamide. J Biosci 36:613–620

    Article  CAS  PubMed  Google Scholar 

  24. Wang H, Lin A, Zhang Z, Chen Y (2001) Expression of porcine growth hormone gene in transgenic rabbits as reported by green fluorescent protein. Anim Biotechnol 12:101–110. https://doi.org/10.1081/ABIO-100108336

    Article  CAS  PubMed  Google Scholar 

  25. Webster NL, Forni M, Bacci ML et al (2005) Multi-transgenic pigs expressing three fluorescent proteins produced with high efficiency by sperm mediated gene transfer. Mol Reprod Dev 72:68–76. https://doi.org/10.1002/mrd.20316

    Article  CAS  PubMed  Google Scholar 

  26. Perry ACF, Wakayama T, Kishikawa H et al (1999) Mammalian transgenesis by intracytoplasmic sperm injection. Science 284:1180–1183. https://doi.org/10.1126/science.284.5417.1180

    Article  CAS  PubMed  Google Scholar 

  27. Moisyadi S, Kaminski JM, Yanagimachi R (2009) Use of intracytoplasmic sperm injection (ICSI) to generate transgenic animals. Comp Immunol Microbiol Infect Dis 32:47–60. https://doi.org/10.1016/j.cimid.2008.05.003

    Article  PubMed  Google Scholar 

  28. Moreira P, Pérez-Cerezales S, Laguna R et al (2016) Transgenic mouse offspring generated by ROSI. J Reprod Dev 62:37–42. https://doi.org/10.1262/jrd.2015-105

    Article  CAS  PubMed  Google Scholar 

  29. Kimura Y, Yanagimachi R, Kuretake S et al (1998) Analysis of mouse oocyte activation suggests the involvement of sperm perinuclear material. Biol Reprod 58:1407–1415

    Article  CAS  PubMed  Google Scholar 

  30. O’Donnell L, O’Bryan MK (2014) Microtubules and spermatogenesis. Semin Cell Dev Biol 30:45–54. https://doi.org/10.1016/j.semcdb.2014.01.003

    Article  CAS  PubMed  Google Scholar 

  31. Katebi S, Esmaeili A, Ghaedi K (2016) Static magnetic field reduced exogenous oligonucleotide uptake by spermatozoa using magnetic nanoparticle gene delivery system. J Magn Magn Mater 402:184–189. https://doi.org/10.1016/j.jmmm.2015.11.057

    Article  CAS  Google Scholar 

  32. Zhang Y, Xi Q, Ding J et al (2012) Production of transgenic pigs mediated by pseudotyped lentivirus and sperm. PLoS One 7:e35335. https://doi.org/10.1371/journal.pone.0035335

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  33. Whittingham DG (1971) Culture of mouse ova. J Reprod Fertil Suppl 14:7–21

    CAS  PubMed  Google Scholar 

  34. Montoliu L (1997) Generation of transgenic mice. A laboratory manual

    Google Scholar 

Download references

Acknowledgments

This work was supported by the Italian Minister of Research and University (FIRB, RBAP06LAHL).

Author information

Author notes

  1. Roberto Giovannoni

    Present address: Department of Biology, University of Pisa, Pisa, Italy

Authors and Affiliations

  1. School of Medicine and Surgery, University of Milano-Bicocca, Monza, Italy

    Marialuisa Lavitrano, Laura Farina, Maria Grazia Cerrito & Roberto Giovannoni

Authors
  1. Marialuisa Lavitrano
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Laura Farina
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Maria Grazia Cerrito
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Roberto Giovannoni
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Marialuisa Lavitrano .

Editor information

Editors and Affiliations

  1. Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Transgenic and Gene-Targeting Facility, University of Kansas Medical Center, Kansas City, KS, USA, Kansas City, KS, USA

    Melissa A. Larson

Rights and permissions

Reprints and permissions

Copyright information

© 2020 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Lavitrano, M., Farina, L., Cerrito, M.G., Giovannoni, R. (2020). Sperm-Mediated Genetic Modifications. In: Larson, M. (eds) Transgenic Mouse. Methods in Molecular Biology, vol 2066. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9837-1_11

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-9837-1_11

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9836-4

  • Online ISBN: 978-1-4939-9837-1

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics

Search

Navigation