Institut Pasteur

Other

Centre National De La Recherche Scientifique

Institut Curie

Charles Babinet

Andre Choulika

Sylvie Robine

Daniel Louvard

Michel Cohen-Tannoudji

Frederic Jaisser

Dominique Morello

The invention relates to methods of introducing a heterologous DNA sequence into a mouse embryonic stem cell wherein the DNA sequence is inserted by homologous recombination into a villin gene/I-SceI hybrid by creating a double strand break with I-SceI meganuclease. Subsequently, the mouse embryonic stem cells can be used to generate a transgenic mouse comprising the heterologous DNA sequence. Additionally, the methods can be used for gene replacement in ovo where a mouse oocyte containing a villin gene/I-SceI hybrid within its genome exists or is first generated. More generally, the methods can be used for the targeted insertion of a heterologous DNA sequence into any cell containing a villin gene/I-SceI hybrid sequence within its genome.

I-Sce I induced gene replacement and gene conversion in embryonic stem cells

This invention relates to a method of constructing a villin gene hybrid by inserting an I-Sce I restriction site next to or within a gene or cDNA encoding a villin protein. The insertion site of the I-Sce I restriction site is chosen as to provide a first downstream part and a second upstream part from the site, containing at least twelve nucleotides of the gene or cDNA encoding the villin protein. Furthermore, the insertion of the restriction permits a high frequency of homologous recombination events. The villin gene hybrid may be used to transfect eukaryotic cells, and particularly, embryonic stem cells.

I Sce I-induced gene replacement and gene conversion in embryonic stem

Tissue cell culture preparations derived from the cells of a transgenic animal are disclosed. These tissue cell preparations are capable of expressing a non-analogous gene product, i.e., a gene product which is not naturally produced by the animal cells. Transhybridomas, i.e., hybridomas and immortalized cells created using cells obtained from transgenic animals, and methods for creating these transhybridomas are also disclosed. In particular, two transhybridomas, A 15.9 and B 6.4, both capable of producing human growth hormone, have been disclosed.

Transhybridomas

Growing community of inventors

Paris, France

Charles Babinet