Compositions and methods for screening cis regulatory elements
AssigneeThe Broad Institute, Inc.
InventorsBinhui Zhao, Benjamin E. Deverman
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Abstract
The invention provides compositions and methods that are useful for screening gene regulatory elements for cell type-specific expression in vivo.
Claims20
A vector comprising a polynucleotide comprising a cis regulatory element and a promoter sequence, each within a region of the polynucleotide defined by two recombinase recognition sites, wherein contacting the polynucleotide with a recombinase forms a mini-circle and stabilizes mRNA transcribed from the polynucleotide in a cell.
A method for screening cis regulatory elements for cell type-specific expression, the method comprising:
- administering to a subject a vector comprising a polynucleotide comprising the following disposed within a region of the polynucleotide defined by two recombinase recognition sites:
- a cis regulatory element, a promoter sequence, a barcode, and an invertible spacer sequence that is disposed within a second set of recombinase recognition sites.
A vector comprising in order from 5′ to 3′:
- a flippase recognition target (FRT) site, a 3′ UTR, a cis regulatory element, a promoter, a (S/W)15VHDB bar code, a lox site, an invertible spacer sequence, a second lox site, and a second flippase recognition target (FRT) site.
A method for screening cis regulatory elements for cell type-specific expression, the method comprising:
- contacting cells with a vector comprising a polynucleotide comprising the following disposed within a region of the polynucleotide defined by two recombinase recognition sites:
- a cis regulatory element, a promoter sequence, a barcode, and an invertible spacer sequence that is disposed within a second set of recombinase recognition sites.
Dependencies are inferred from the claim text.