Predetermined chromosomal deletion encompassing the Nf-1 gene.
Complex chromosomal rearrangements (deletions, inversions, translocations) are a hallmark of human tumour cells. Yet, the generation of animal models for gross chromosomal abnormalities still presents a formidable challenge. Here, we describe a versatile procedure for chromosomal engineering that was used to generate an ES cell line with a megabase deletion encompassing the tumour suppressor gene neurofibromatosis-1 (Nf-1) on mouse chromosome 11, which is often deleted in tumours of neural crest origin. Homologous recombination into sites flanking Nf-1 was used to introduce artificial sequences (triple-helix, loxP, vector backbone) that can be employed for in vitro recovery of intervening sequences or the generation of in vivo deletions. This strategy may be developed into a scheme by which large chromosomal regions with precisely defined end points may be excised from mammalian cells and reintroduced after suitable in vitro modification.
Schlake
T
T
Department of Developmental Immunology, Max-Planck-Institute for Immunobiology, Stuebeweg 51, D-79108 Freiburg, Germany.
Schupp
I
I
Kutsche
K
K
Mincheva
A
A
Lichter
P
P
Boehm
T
T
D000595:Amino Acid Sequence; D000818:Animals; D015139:Blotting, Southern; D002460:Cell Line; D002872:Chromosome Deletion; D003001:Cloning, Molecular / Q000379:methods; D015321:Gene Rearrangement; D005821:Genetic Techniques; D049452:Green Fluorescent Proteins; D019426:Integrases / Q000235:genetics; D008164:Luminescent Proteins / Q000235:genetics / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D025542:Neurofibromin 1; D016133:Polymerase Chain Reaction; D011506:Proteins / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D018741:Replication Origin / Q000235:genetics; D013234:Stem Cells; D014764:Viral Proteins
D008164:Luminescent Proteins; D025542:Neurofibromin 1; D011506:Proteins; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D049452:Green Fluorescent Proteins; C045073:Cre recombinase; D019426:Integrases