Modified es cells and es cells-specific gene
OwnerHerve Acloque, Anne-Marie Birot +3inventors
InventorsHerve Acloque, Anne-Marie Birot, Valerie Risson, Bertrand Pain, Jacques Samarut
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Abstract
The invention concerns modified avian ES cells, specifically expressing an exogenous gene when they have a pluripotent character. The invention also concerns a nucleic acid and a polypeptide specifically expressed in pluripotent avian cells, and methods for detecting the pluripotent character of cells using said nucleic acid and polypeptide.
Claims45
A purified or isolated nucleic acid characterized in that it comprises a nucleic acid sequence chosen from the group of following sequences:
- a) SEQ ID No. 1, or the fragment corresponding to nucleotides 1409-2878 of SEQ ID No. 1;
- b) the sequence of a fragment of at least 15 consecutive nucleotides of a sequence chosen from SEQ ID No. 1, in particular the fragment corresponding to nucleotides 3111-3670 of SEQ ID No. 1;
- c) a nucleic acid sequence having a percentage identity of at least 80%, after the optimal alignment, with a sequence defined in a) or b), said sequence not being defined by nucleotides 2308-2927 or 3094-3753 of SEQ ID No. 1;
- d) a nucleic acid sequence which hybridizes, under high stringency conditions, with a nucleic acid sequence defined in a) or b), said sequence not being defined by nucleotides 2308-2927 or 3094-3753 of SEQ ID No. 1;
- e) the complementary sequence or the RNA sequence corresponding to a sequence as defined in a), b), c) or d).
A purified or isolated nucleic acid, characterized in that it encodes a polypeptide which has a continuous fragment of at least 200 amino acids of the protein SEQ ID No. 2.
An isolated polypeptide, characterized in that it comprises a polypeptide chosen from:
- a) a polypeptide corresponding to SEQ ID No. 2;
- b) a variant polypeptide of a polypeptide of sequence defined in a);
- c) a polypeptide homologous to a polypeptide defined in a) or b), comprising at least 80% homology with said polypeptide of a);
- d) a fragment of at least 15 consecutive amino acids of a polypeptide defined in a), b) or c);
- e) a biologically active fragment of a polypeptide defined in a), b) or c).
A method for determining the pluripotent nature of an avian ES cell, characterized in that the presence of a product of expression of the gene corresponding to SEQ ID No. 1, or of the mRNA of SEQ ID No. 1, is determined.
A method for screening for a substance capable of restoring the pluripotent nature of differentiated cells, characterized in that it comprises the following steps:
- a) maintaining differentiated cells in a suitable culture medium;
- b) replacing said culture medium with a medium which makes it possible to maintain a pluripotent phenotype and which contains said substance to be tested;
- c) determining the restoration of the pluripotent nature of said cells by the expression of the protein SEQ ID No. 2 or of the exogenous gene, in said cells.
The use of a nucleic acid corresponding to nucleotides 3111-3670 of SEQ ID No. 1, as a promoter of a gene of interest, for specific expression of said gene of interest in avian pluripotent cells.
Dependencies are inferred from the claim text.