Skip to main content

Dendrimer Mediated Transfer of Engineered Chromosomes

  • Protocol
  • First Online:
Book cover Mammalian Chromosome Engineering

Part of the book series: Methods in Molecular Biology ((MIMB,volume 738))

Abstract

Gene therapy encounters important problems such as insertional mutagenesis caused by the integration of viral vectors. These problems could be circumvented by the use of mammalian artificial chromosomes (MACs) that are unique and high capacity gene delivery tools. MACs were delivered into various target cell lines including stem cells by microcell-mediated chromosome transfer (MMCT), microinjection, and cationic lipid and dendrimer mediated transfers. MACs were also cleansed to more than 95% purity before transfer with an expensive technology. We present here a method by which MACs can be delivered into murine embryonic stem (ES) cells with a nonexpensive, less tedious, but still efficient way.

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

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 139.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
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. Hadlaczky G. Satellite DNA-based artificial chromosomes for use in gene therapy. Curr Opin Mol Ther. 2001;3:125–32.

    PubMed  CAS  Google Scholar 

  2. Lindenbaum M, Perkins E, Csonka E, Fleming E, Garcia L, Greene A, et al. A mammalian artificial chromosome engineering system (ACE System) applicable to biopharmaceutical protein production, transgenesis and gene-based cell therapy. Nucleic Acids Res. 2004;32:e172.

    Article  PubMed  Google Scholar 

  3. Schor SL, Johnson RT, Mullinger AM. Perturbation of mammalian cell division. II. Studies on the isolation and characterization of human mini segregant cells. J Cell Sci. 1975;19:281–303.

    PubMed  CAS  Google Scholar 

  4. Fournier RE, Ruddle FH. Microcell-mediated transfer of murine chromosomes into mouse, Chinese hamster, and human somatic cells. Proc Natl Acad Sci USA. 1977;74:319–23.

    Article  PubMed  CAS  Google Scholar 

  5. Meaburn KJ, Parris CN, Bridger JM. The manipulation of chromosomes by mankind: the uses of microcell-mediated chromosome transfer. Chromosoma. 2005;114:263–74.

    Article  PubMed  Google Scholar 

  6. Doherty AMO, Fisher EMC. Microcell-mediated chromosome transfer (MMCT): small cells with huge potential. Mamm Genome. 2003;14:583–92.

    Article  PubMed  Google Scholar 

  7. Kakeda M, Hiratsuka M, Nagata K, Kuroiwa Y, Kakitani M, Katoh M, et al. Human artificial chromosome (HAC) vector provides long-term therapeutic transgene expression in normal human primary fibroblasts. Gene Ther. 2005;12:852–6.

    Article  PubMed  CAS  Google Scholar 

  8. Yamada H, Kunisato A, Kawahara M, Tahimic CG, Ren X, Ueda H, et al. Exogenous gene expression and growth regulation of hematopoietic cells via a novel human artificial chromosome. J Hum Genet. 2006;51:147–50.

    Article  PubMed  CAS  Google Scholar 

  9. Co DO, Borowski AH, Leung JD, van der Kaa J, Hengst S, Platenburg GJ, et al. Generation of transgenic mice and germline transmission of a mammalian artificial chromosome introduced into embryos by pronuclear microinjection. Chromosome Res. 2000;8:183–91.

    Article  PubMed  CAS  Google Scholar 

  10. Monteith DP, Leung JD, Borowski AH, Co DO, Praznovszky T, Jirik FR, et al. Pronuclear microinjection of purified artificial chromosomes for generation of transgenic mice: pick-and-inject technique. Methods Mol Biol. 2004;240:227–42.

    PubMed  CAS  Google Scholar 

  11. Vanderbyl S, MacDonald GN, Sidhu S, Gung L, Telenius A, Perez C, et al. Transfer and stable transgene expression of a mammalian artificial chromosome into bone marrow-derived human mesenchymal stem cells. Stem Cells. 2004;22:324–33.

    Article  PubMed  CAS  Google Scholar 

  12. Vanderbyl SL, Sullenbarger B, White N, Perez CF, MacDonald GN, Stodola T, et al. Transgene expression after stable transfer of a mammalian artificial chromosome into human hematopoietic cells. Exp Hematol. 2005;33:1470–6.

    Article  PubMed  CAS  Google Scholar 

  13. de Jong G, Telenius A, Vanderbyl S, Meitz A, Drayer J. Efficient in-vitro transfer of a 60-Mb mammalian artificial chromosome into murine and hamster cells using cationic lipids and dendrimers. Chromosome Res. 2001;9:475–85.

    Article  PubMed  Google Scholar 

  14. Katona RL, Sinkó I, Holló G, Szucs KS, Praznovszky T, Kereső J, et al. A combined artificial chromosome-stem cell therapy method in a model experiment aimed at the treatment of Krabbe’s disease in the Twitcher mouse. Cell Mol Life Sci. 2008;65:3830–8.

    Article  PubMed  CAS  Google Scholar 

  15. Nagy A, Rossant J, Nagy R, Abramow-Newerly W, Roder JC. Derivation of completely cell culture-derived mice from early-passage embryonic stem cells. Proc Natl Acad Sci USA. 1993;90:8424–8.

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Robert L. Katona .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2011 Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Katona, R.L. (2011). Dendrimer Mediated Transfer of Engineered Chromosomes. In: Hadlaczky, G. (eds) Mammalian Chromosome Engineering. Methods in Molecular Biology, vol 738. Humana Press. https://doi.org/10.1007/978-1-61779-099-7_11

Download citation

  • DOI: https://doi.org/10.1007/978-1-61779-099-7_11

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-098-0

  • Online ISBN: 978-1-61779-099-7

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics