RNA-Guided Transcriptional Regulation

A method of modulating expression of a target nucleic acid in a cell comprising providing to the cell a guide RNA complementary to the target nucleic acid sequence including a transcriptional activator or repressor domain connected thereto for modulating target nucleic acid expression in vivo,

• providing to the cell a nuclease null Cas9 protein that interacts with the guide RNA and binds to the target nucleic acid sequence in a site specific manner,
• wherein the guide RNA including the transcriptional activator or repressor domain connected thereto and the Cas9 protein co-localize to the target nucleic acid sequence and wherein the transcriptional activator or repressor domain modulates expression of the target nucleic acid.

The method of claim 2

• wherein the guide RNA including the transcriptional activator or repressor domain connected thereto is provided to the cell by introducing to the cell a nucleic acid encoding the guide RNA including the transcriptional activator or repressor domain as a fusion protein,
• wherein the Cas9 protein is provided to the cell by introducing to the cell a nucleic acid encoding the Cas9 protein, and
• wherein the cell expresses the guide RNA including the transcriptional activator or repressor domain as a fusion protein and the Cas9 protein.

The method of claim 1 wherein the cell is a eukaryotic cell.

The method of claim 1 wherein the cell is a yeast cell, a plant cell or a mammalian cell.

The method or claim 1 wherein the cell is a human cell.

The method of claim 1 wherein the guide RNA is between about 10 to about 250 nucleotides.

The method of claim 1 wherein the guide RNA is between about 20 to about 100 nucleotides.

The method of claim 1 wherein the guide RNA is between about 100 to about 250 nucleotides.

The method of claim 1 wherein the target nucleic acid is genomic DNA, mitochondrial DNA, viral DNA or exogenous DNA.

A method of modulating expression of a target nucleic acid in a cell comprising

• providing to the cell a guide RNA complementary to viral DNA including the target nucleic acid sequence, wherein the guide RNA includes a transcriptional activator or repressor domain connected thereto for modulating target nucleic acid expression in vivo,
• providing to the cell a nuclease null Cas9 protein that interacts with the guide RNA and binds to the target nucleic acid sequence in a site specific manner,
• wherein the guide RNA including the transcriptional activator or repressor domain connected thereto and the Cas9 protein co-localize to the target nucleic acid sequence and wherein the transcriptional activator or repressor domain modulates expression of the target nucleic acid.

The method of claim 10

• wherein the guide RNA including the transcriptional activator or repressor domain connected thereto is provided to the cell by introducing to the cell a nucleic acid encoding the guide RNA including the transcriptional activator or repressor domain as a fusion protein,
• wherein the Cas9 protein is provided to the cell by introducing to the cell a nucleic acid encoding the Cas9 protein, and
• wherein the cell expresses the guide RNA including the transcriptional activator or repressor domain as a fusion protein and the Cas9 protein.

The method of claim 10 wherein the cell is a eukaryotic cell.

The method of claim 10 wherein the cell is a yeast cell, a plant cell or a mammalian cell.

The method or claim 10 wherein the cell is a human cell.

The method of claim 10 wherein the guide RNA is between about 10 to about 250 nucleotides.

The method of claim 10 wherein the guide RNA is between about 20 to about 100 nucleotides.

The method of claim 10 wherein the guide RNA is between about 100 to about 250 nucleotides.