Use of regulatory sequences for specific, transient expression in neuronal determined cells

摘要

The present invention relates to the use of regulatory sequences for mediating specific, early transient expression in proliferative neuronal determined cells. Furthermore, the uses of recombinant nucleic acid molecules comprising said defined regulatory sequences for mediating specific, early transient expression in proliferative neuronal determined cells as well as for the generation of non-human transgenic organisms and/or host cells are disclosed. In addition, the invention provides for transgenic non-human animals and/or host cells comprising said regulatory sequences and/or recombinant nucleic acid molecules. The invention also describes methods for the preparation of such vectors, host cells and transgenic non-human animals as well as methods for the detection and/or isolation of neuronal determined cells. Additionally, methods for screening of compounds capable of regulating neuronal determined cell activity, neurogenesis, stimulating proliferation of neuronally committed precursor cells and/or neuronal differentiation are provided and the invention also relates to methods for the detection and analysis of neuronal differentiation, neuronal migration and/or neuronal determination processes. Finally, the invention relates to diagnostic and pharmaceutical compositions comprising the regulatory sequences, recombinant nucleic acid molecules, host-cells or isolated neuronal determined cells described herein.

主权项

1. A method for the detection of (a) proliferative neuronal determined cells comprising the steps of:
expressing in a plurality of cells a recombinant nucleic acid molecule comprising a heterologous nucleotide sequence to be expressed, said sequence under the control of a regulatory sequence selected from the group consisting of:
(a) regulatory sequences comprising the nucleotide sequence shown in SEQ ID NO: 1, as shown in SEQ ID NO: 2, as shown in SEQ ID NO: 3 or as shown in SEQ ID NO: 4;(b) regulatory sequences comprising the nucleotide sequence contained in the insertion of clone DSM 15111 and obtainable by amplification using two oligonucleotides having the sequences indicated under SEQ ID NO: 9 and SEQ ID NO: 10;(c) regulatory sequences comprising at least one nucleotide sequence of SEQ ID NO: 1 from position 1166 to 1746, from position 1166 to 2049, from position 1785 to 1843 or from position 1953 to 2775;(d) regulatory sequences comprising at least one nucleotide sequence of SEQ ID NO: 2 from position 529 to 1079, from position 529 to 1390, from position 1118 to 1175 or from position 1291 to 2137; regulatory sequences comprising a nucleotide sequence which is at least 75% identical to a sequence as defined in (a) to (d) or which comprises a nucleotide sequence which is at least 78% identical to the nucleotide sequence as shown in SEQ ID NO: 1 from position 1166 to 1746 or from position 1166 to 2049 or to the nucleotide sequence shown in SEQ ID NO: 2 from position 529 to 1079 or from position 529 to 1390, which comprises a nucleotide sequence which is at least 82% identical to the nucleotide sequence as shown in SEQ ID NO: 1 from position 1785 to 1843 or to the nucleotide sequence as shown in SEQ ID NO: 2 from position 1118 to 1175 or which comprises a nucleotide sequence which is at least 75% identical to the nucleotide sequence as shown in SEQ ID NO: 1 from position 1953 to 2775 or to the nucleotide sequence as shown in SEQ ID NO: 2 from position 1291 to 2137; and(f) regulatory sequences comprising a nucleotide sequence which hybridizes under stringent conditions with a complementary strand of the regulatory sequence as defined in (a) to (e) and which provides early, transient expression of a heterologous nucleotide sequence in proliferative neuronal determined cells; and selecting cells which express a heterologous nucleotide sequence under the control of the regulatory sequence.