Compositions and methods for altering gene expression
AssigneeWhitehead Institute for Biomedical Research
InventorsJill M. Dowen, Zi Peng Fan, Denes Hnisz, Tong Ihn Lee, Richard A. Young
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Abstract
Provided herein are improved compositions and methods for the directed control of gene expression.
Claims8
A method of decreasing expression of a murine Nanog gene, the method comprising contacting a murine embryonic stem cell with a gene modulatory composition , wherein
- the Nanog gene is located in an insulated neighborhood (IN) formed by looping of two CTCF interaction sites occupied by cohesin on a chromosome in the cell,
- and the gene modulatory composition is a CRISPR/Cas9 system comprising Cas9 and a first guide RNA complementary to SEQ ID NO:
- 8 and a second guide RNA complementary to SEQ ID NO:
- 9, wherein the gene modulatory composition deletes the sequence of one of the CTCF interaction sites and disrupts binding of CTCF protein, and wherein expression of the Nanog gene is decreased by about 40%.
A method of decreasing expression of a Nanog gene in a murine embryonic stem cell in cell culture, the method comprising deleting a DNA sequence comprising one of two CTCF interaction sites that flank the Nanog gene on a chromosome in the cell using a CRISPR/Cas9 system, thereby generating a deleted CTCF interaction site and disrupting binding of CTCF protein to the deleted CTCF interaction site,
- wherein the Nanog gene is located in an insulated neighborhood (IN) formed by looping of the two CTCF interaction sites occupied by cohesin on the chromosome;
- and
- wherein the CRISPR/Cas9 system comprises Cas9 and a first guide RNA complementary to SEQ ID NO:
- 8 and a second guide RNA complementary to SEQ ID NO:
- 9, and wherein expression of the Nanog gene is decreased by about 40%.
Dependencies are inferred from the claim text.