Methods of altering gene expression by perturbing transcription factor multimers that structure regulatory loops
The invention relates to methods of modulating the expression of one or more genes in a cell by modulating the multimerization of a transcription factor and/or modulating the formation of enhancer-promoter DNA loops, and thereby modulating the expression of the one or more genes. The invention also relates to treating diseases and conditions involving aberrant gene expression by modulating the multimerization of a transcription factor and/or modulating the formation of enhancer-promoter DNA loops. The invention also relates to methods for screening for compounds that modulate expression of one or more genes in a cell.
1. A method of decreasing expression of one or more genes in a cell, comprising contacting the cell with a composition for decreasing binding of Yin Yang 1 (YY1) to a YY1 binding site, wherein the composition comprises a site-specific DNA binding domain (DBD) that binds a promoter or an enhancer of the one or more genes and an effector domain having DNA methylation activity, or one or more nucleic acids encoding the site-specific DBD and/or effector domain, and wherein the YY1 binding site is in the promoter or the enhancer of the one or more genes, thereby decreasing expression of the one or more genes.
2. The method of claim 1 , wherein the site-specific DBD comprises a zinc finger (ZF).
3. The method of claim 1 , wherein the site-specific DBD comprises a transcription activator-like effector (TALE).
4. The method of claim 1 , wherein the site-specific DBD comprises a catalytically inactive nuclease and a guide sequence that binds the target sequence.
5. The method of claim 4 , wherein the catalytically inactive nuclease is a catalytically inactive Cas protein.
6. The method of claim 1 , wherein the site-specific DBD is operably linked to the effector domain.
7. The method of claim 1 , wherein the composition comprises a nucleic acid comprising a nucleotide sequence encoding the site-specific DBD operably linked to the effector domain.
8. The method of claim 7 , wherein the composition comprises the nucleic acid formulated in a lipid nanoparticle.
9. The method of claim 1 , wherein the YY1 binding site is in the enhancer of the one or more genes.
10. The method of claim 1 , wherein the YY1 binding site is in the promoter of the one or more genes.
11. The method of claim 1 , wherein the effector domain having DNA methylation activity increases DNA methylation of the one or more genes in a region of about 25 bases about 50 bases, about 100 bases, about 200 bases, about 300 bases, about 400 bases, about 500 bases, about 600 bases, about 700 bases, about 800 bases, about 900 bases, or about 1000 bases spanning the YY1 binding site.
12. The method of claim 1 , wherein the one or more genes comprises an oncogene.
13. The method of claim 1 , wherein binding of YY1 to the YY1 binding site is decreased compared to a control cell not contacted with the composition.
14. A method of decreasing expression of one or more genes in a cell, comprising contacting the cell with a composition for inhibiting formation of an enhancer-promoter DNA loop comprising the one or more genes, wherein the formation of the enhancer-promoter DNA loop is YY1 dependent, wherein the composition comprises a polypeptide for decreasing YY1 multimerization, or one or more nucleic acids encoding the polypeptide.
15. The method of claim 14 , wherein the one or more genes comprises an oncogene.
16. The method of claim 14 , wherein the composition comprises the polypeptide.
17. The method of claim 14 , wherein the composition comprises one or more nucleic acids encoding the polypeptide.
18. The method of claim 17 , wherein the composition comprises the one or more nucleic acids formulated in a lipid nanoparticle.