PMID: 9647640Jul 1, 1998Paper

Rescue of targeted regions of mammalian chromosomes by in vivo recombination in yeast

Genome Research
N KouprinaM Koi

Abstract

In contrast to other animal cell lines, the chicken pre-B cell lymphoma line, DT40, exhibits a high level of homologous recombination, which can be exploited to generate site-specific alterations in defined target genes or regions. In addition, the ability to generate human/chicken monochromosomal hybrids in the DT40 cell line opens a way for specific targeting of human genes. Here we describe a new strategy for direct isolation of a human chromosomal region that is based on targeting of the chromosome with a vector containing a yeast selectable marker, centromere, and an ARS element. This procedure allows rescue of the targeted region by transfection of total genomic DNA into yeast spheroplasts. Selection for the yeast marker results in isolation of chromosome sequences in the form of large circular yeast artificial chromosomes (YACs) up to 170 kb in size containing the targeted region. These YACs are generated by homologous recombination in yeast between common repeated sequences in the targeted chromosomal fragment. Alternatively, the targeted region can be rescued as a linear YACs when a YAC fragmentation vector is included in the yeast transformation mixture. Because the entire isolation procedure of the chromosomal region...Continue Reading

References

Sep 15, 1992·Proceedings of the National Academy of Sciences of the United States of America·H ShizuyaM Simon
Jun 1, 1990·Proceedings of the National Academy of Sciences of the United States of America·S G GrantD Hanahan
Aug 1, 1994·Human Molecular Genetics·K E BrownW R Brown
Jun 7, 1994·Proceedings of the National Academy of Sciences of the United States of America·G Bepler, M A Garcia-Blanco
Jan 9, 1996·Proceedings of the National Academy of Sciences of the United States of America·V LarionovM A Resnick
Nov 26, 1996·Proceedings of the National Academy of Sciences of the United States of America·V LarionovM A Resnick
Nov 29, 1996·Cell·J V MoranH H Kazazian
Jan 7, 1997·Proceedings of the National Academy of Sciences of the United States of America·P Ross-MacdonaldM Snyder
May 1, 1997·Nature Genetics·D M SassamanH H Kazazian
Jan 1, 1997·Cytogenetics and Cell Genetics·M KoiJ C Barrett
Jul 8, 1997·Proceedings of the National Academy of Sciences of the United States of America·V LarionovM A Resnick
May 16, 1998·Proceedings of the National Academy of Sciences of the United States of America·N KouprinaV Larionov

❮ Previous
Next ❯

Citations

Mar 10, 2010·Nucleic Acids Research·Gwynedd A BendersClyde A Hutchison
Mar 28, 2003·Human Gene Therapy·Craig A HokansonMichael J Mendez

❮ Previous
Next ❯

Related Concepts

Related Feeds

B-Cell Lymphoma

B-cell lymphomas include lymphomas that affect B cells. This subtype of cancer accounts for over 80% of non-Hodgkin lymphomas in the US. Here is the latest research.

Artificial Chromosomes

Artificial chromosomes are genetically engineered chromosomes derived from the DNA of a species. Discover the latest research on artificial chromosomes here.