CONSTRUCT FOR EPIGENETIC MODIFICATION AND ITS USE IN THE SILENCING OF GENES
The present invention concerns a construct for epigenomic modification of genes that includes the following components: a) a Krüppel-associated box zinc finger protein or homologous, b) a DNA region capable of binding to the target gene or homologous, c) a human DNA methyltransferase DNMT3A or homologous and d) a murine DNA methyltransferase Dnmt3L or homologous whereby components a), b), c) and d) are linked to each other either directly or via at least one linker. The construct is a designer epigenome modifier which can be used to silence genes coding for a protein in leukocytes which avoids the internalization of HI viruses in immune cells.
1 . A construct for targeted epigenomic modification of genes comprising the following components:
a) a Krüppel-associated box zinc finger protein or homologous,
b) a DNA region capable of binding to the target gene or homologous,
c) a human DNA methyltransferase DNMT3A or homologous and
d) a murine DNA methyltransferase Dnmt3L or homologous
whereby components a), b), c) and d) are linked to each other either directly or via at least one linker.
2 . The construct according to claim 1 , characterized in that the single components are proteins and/or peptides.
3 . The construct according to claim 1 , characterized in that the construct is a nucleic acid based construct coding for the components a)-d) and at least a linker.
4 . The construct according to claim 3 , characterized in that the nucleic acid construct is contained within a vector.
5 . The construct according to claim 4 , characterized in that the vector is a vector derived from a lentivirus, an adenovirus or an adeno-associated virus.
6 . The construct according to claim 3 , characterized in that it is an mRNA molecule which may additionally comprise further components selected from the group consisting of a 7-methylguanosin cap (or an artificial cap analogue) at the 5′-end, a non-coding region at the 5′-end, a non-coding region at the 3′-end and/or a polyA tail at the 3′-end.
7 . The construct according to claim 3 , characterized in that the nucleic acid is a DNA.
8 . A method for silencing a gene or genes of interest comprising the steps of:
(a) introducing into target cells a construct for targeted epigenomic modification of genes according to claim 1 ;
(b) silencing the gene or genes of interest in said target cells via epigenetic modification.
9 . The method according to claim 8 , characterized in that the target cells are primary cells.
10 . The method according to claim 8 , wherein step (a) is performed by introducing a polynucleotide RNA molecule coding for said construct for targeted epigenomic modification of genes according to claim 1 into said target cells.
11 . The method according to claim 10 , wherein step (a) is performed ex vivo.
12 . The method according to claim 8 , wherein the chromatin architecture of said target cells is dissected.
13 . The method according to claim 8 , wherein said method is applied to the treatment of a disease or disorder in a patient in need thereof.
14 . The method according to claim 13 , wherein said target cells are isolated lymphocytes, said genes of interest being silenced are genes required for the expression of a co-receptor required by an immune deficiency virus to enter into said cells and said disease or disorder being treated is an infection with said immune deficiency virus.
15 . The method according to claim 13 , wherein said target cells are isolated leukocytes, said genes of interest being silenced are genes required for the expression of a receptor required by human virus to enter into said cells and said disease or disorder being treated is an infection with said human virus.
16 . The method according to claim 13 , wherein said target cells are isolated human cells, said genes of interest being silenced are genes required for the multiplication of a human pathogen and said disease or disorder being treated is an infection with said human pathogen.