Genome Research scroll

Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
QUICK SEARCH:   [advanced]


     

This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Right arrow Citation Map
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrow reprints & permissions
Citing Articles
Right arrow Citing Articles via HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Voet, T.
Right arrow Articles by Marynen, P.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Voet, T.
Right arrow Articles by Marynen, P.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us   Add to Digg   Add to Reddit   Add to Technorati  
What's this?

Vol. 11, Issue 1, 124-136, January 2001

METHODS
Efficient Male and Female Germline Transmission of a Human Chromosomal Vector in Mice

Thierry Voet,1,4 Joris Vermeesch,1,2,4 An Carens,1 Joachim Dürr,3 Charlotte Labaere,1 Hein Duhamel,1 Guido David,3 and Peter Marynen1,5,6

1 Human Genome Laboratory, Leuven, Belgium; 2 Aventis CropScience, Ghent; 3 Laboratory for Glycobiology and Developmental Genetics, Center for Human Genetics, Flanders Interuniversity Institute for Biotechnology, University of Leuven, B-3000 Leuven, Belgium

A small accessory chromosome that was mitotically stable in human fibroblasts was transferred into the hprt- hamster cell line CH and developed as a human chromosomal vector (HCV) by the introduction of a selectable marker and the 3' end of an HPRT minigene preceded by a loxP sequence. This HCV is stably maintained in the hamster cell line. It consists mainly of alphoid sequences of human chromosome 20 and a fragment of human chromosome region 1p22, containing the tissue factor gene F3. The vector has an active centromere, and telomere sequences are lacking. By transfecting a plasmid containing the 5' end of HPRT and a Cre-encoding plasmid into the HCV+ hamster cell line, the HPRT minigene was reconstituted by Cre-mediated recombination and expressed by the cells. The HCV was then transferred to male mouse R1-ES cells and it did segregate properly. Chimeras were generated containing the HCV as an independent chromosome in a proportion of the cells. Part of the male and female offspring of the chimeras did contain the HCV. The HCV+ F1 animals harbored the extra chromosome in >80% of the cells. The HCV was present as an independent chromosome with an active centromere and the human F3 gene was expressed from the HCV in a human-tissue-specific manner. Both male and female F1 mice did transmit the HCV to F2 offspring as an independent chromosome with properties similar to the original vector. This modified small accessory chromosome, thus, shows the properties of a useful chromosomal vector: It segregates stably as an independent chromosome, sequences can be inserted in a controlled way and are expressed from the vector, and the HCV is transmitted through the male and female germline in mice.

4 These authors contributed equally to this work.

5 Present address: Center for Human Genetics, Flanders Interuniversity Institute for Biotechnology, University of Leuven, Campus Gasthuisberg, Herestraat 49, B-3000 Leuven, Belgium.

6 Corresponding author.

11:124-136 ©2001 by Cold Spring Harbor Laboratory Press  ISSN 1088-9051/01 $5.00
Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us   Add to Digg Digg   Add to Reddit Reddit   Add to Technorati Technorati    What's this?


This article has been cited by other articles:


Home page
 Stem CellsHome page
M. Paulis, M. Bensi, D. Orioli, C. Mondello, G. Mazzini, M. D'Incalci, C. Falcioni, E. Radaelli, E. Erba, E. Raimondi, et al.
Transfer of a Human Chromosomal Vector from a Hamster Cell Line to a Mouse Embryonic Stem Cell Line
Stem Cells, October 1, 2007; 25(10): 2543 - 2550.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
N. Suzuki, K. Nishii, T. Okazaki, and M. Ikeno
Human Artificial Chromosomes Constructed Using the Bottom-up Strategy Are Stably Maintained in Mitosis and Efficiently Transmissible to Progeny Mice
J. Biol. Chem., September 8, 2006; 281(36): 26615 - 26623.
[Abstract] [Full Text] [PDF]

Home page
 J. Biol. Chem.Home page
L. H. Wong, R. Saffery, M. A. Anderson, E. Earle, J. M. Quach, A. J. Stafford, K. J. Fowler, and K. H. A. Choo
Analysis of Mitotic and Expression Properties of Human Neocentromere-based Transchromosomes in Mice
J. Biol. Chem., February 4, 2005; 280(5): 3954 - 3962.
[Abstract] [Full Text] [PDF]

Home page
 Nucleic Acids ResHome page
M. Lindenbaum, E. Perkins, E. Csonka, E. Fleming, L. Garcia, A. Greene, L. Gung, G. Hadlaczky, E. Lee, J. Leung, et al.
A mammalian artificial chromosome engineering system (ACE System) applicable to biopharmaceutical protein production, transgenesis and gene-based cell therapy
Nucleic Acids Res., December 7, 2004; 32(21): e172 - e172.
[Abstract] [Full Text] [PDF]

Home page
 Arterioscler. Thromb. Vasc. Bio.Home page
G. Schabbauer, M. Tencati, B. Pedersen, R. Pawlinski, and N. Mackman
PI3K-Akt Pathway Suppresses Coagulation and Inflammation in Endotoxemic Mice
Arterioscler. Thromb. Vasc. Biol., October 1, 2004; 24(10): 1963 - 1969.
[Abstract] [Full Text] [PDF]

Home page
 J. Cell Biol.Home page
T. Voet, B. Liebe, C. Labaere, P. Marynen, and H. Scherthan
Telomere-independent homologue pairing and checkpoint escape of accessory ring chromosomes in male mouse meiosis
J. Cell Biol., September 1, 2003; 162(5): 795 - 808.
[Abstract] [Full Text] [PDF]


Home Help [Feedback] [For Subscribers] [Archive] [Search] [Contents]
Genes Dev. Learn. Mem.
Protein Science RNA Genome Res.