Chimeric enzyme for promoting targeted integration of foreign DNA into a host genome
AssigneeFox Chase Cancer Center
InventorsRichard A. Katz, Anna Marie Skalka
Life of this patent
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Abstract
The present invention provides novel chimeric and fusion proteins useful for facilitating site-specific integration of foreign DNA into a host genome. The chimeric enzymes of the invention comprise a DNA binding moiety fused to a retroviral integrase moiety, preferably at the precise N- or C-terminus. Nucleic acids encoding these fusion proteins can be incorporated into standard retroviral vectors, or can be provided as purified proteins. They are capable of exerting the activities of a wildtype retroviral integrase, including processing retroviral DNA termini, nicking double-stranded DNA and integrating a DNA molecule with processed retroviral termini into another DNA strand.
Claims5
A nucleic acid molecule encoding a chimeric enzyme comprising a first segment derived from a first source operably linked to a second segment derived from a second source, said second source being different from said first source, said first segment encoding a DNA binding moiety and said second segment encoding an integrase moiety derivable from a retroelement, said integrase moiety having a carboxy terminus and an amino terminus, said amino terminus having a ZF domain, wherein said integrase moiety is truncated from said amino terminus so as to remove part or all of said ZF domain, and said DNA biding moiety is appended to said integrase moiety at said truncated amino terminus, said enzyme binding specifically to a targeted host DNA molecule having a characteristic determinant recognized by said DNA binding moiety, and possessing integrase activity, whereby said enzyme catalyzes site-specific integration of foreign DNA into a host genome sequence.
Dependencies are inferred from the claim text.