Compositions and methods for epigenome editing
Disclosed herein are CRISPR/Cas9-based gene activation systems that include a fusion protein of a Cas9 protein and a protein having histone acetyltransferase activity, and methods of using said systems.
1. An isolated polynucleotide encoding a fusion protein comprising two heterologous polypeptide domains,
wherein the first polypeptide domain comprises a Clustered Regularly Interspaced Short Palindromic Repeats associated (Cas) protein,
wherein the second polypeptide domain comprises a p300 histone acetyltransferase effector domain, and
wherein the fusion protein comprises the polypeptide sequence of SEQ ID NO: 140, SEQ ID NO: 141, or SEQ ID NO: 149.
2. The isolated polynucleotide of claim 1 , wherein the fusion protein activates transcription of a target gene by activating a regulatory element.
3. The isolated polynucleotide of claim 2 , wherein the regulatory element comprises a promoter of the target gene.
4. A vector comprising the isolated polynucleotide of claim 1 .
5. A kit comprising the isolated polynucleotide of claim 1 .
6. An isolated cell comprising the isolated polynucleotide of claim 1 .
7. An isolated polynucleotide encoding a fusion protein comprising two heterologous polypeptide domains,
wherein the first polypeptide domain comprises a Clustered Regularly Interspaced Short Palindromic Repeats associated (Cas) protein,
wherein the second polypeptide domain comprises a p300 histone acetyltransferase effector domain, and
wherein the first polypeptide domain comprises the polypeptide sequence of SEQ ID NO: 10 and the second polypeptide domain comprises the polypeptide sequence of SEQ ID NO: 3, or the first polypeptide domain comprises the polypeptide sequence of SEQ ID NO: 1 and the second polypeptide domain comprises the polypeptide sequence of SEQ ID NO: 2.
8. The isolated polynucleotide of claim 7 , wherein the fusion protein activates transcription of a target gene by activating a regulatory element.
9. The isolated polynucleotide of claim 8 , wherein the regulatory element comprises a promoter of the target gene.
10. The isolated polynucleotide of claim 7 , further comprising a linker connecting the first polypeptide domain to the second polypeptide domain.
11. A vector comprising the isolated polynucleotide of claim 7 .
12. A kit comprising the isolated polynucleotide of claim 7 .
13. An isolated cell comprising the isolated polynucleotide of claim 7 .