Multiple domain proteins
Described herein are methods and compositions for generating and using fusion proteins.
1. A fusion protein comprising two heterologous polypeptide domains connected by a linker, wherein a first polypeptide domain is configured to associate with a nucleic acid, and a second polypeptide domain has RNA cleavage activity;
wherein the first polypeptide domain is:
SEQ ID NO: 7 (pumilio); and
wherein the second polypeptide domain is a dicer endoribonuclease, wherein the dicer endoribonuclease comprises two RNase III domains and a PAZ domain, and wherein the dicer endoribonuclease cleaves double-stranded RNA and pre-microRNA.
2. A isolated cell comprising the fusion protein of claim 1 .
3. A cell-free system comprising the fusion protein of claim 1 .
4. An in vitro method for modifying a target nucleic acid, comprising contacting the target nucleic acid with the fusion protein of claim 1 under modification conditions, whereby the target nucleic acid is modified by the fusion protein.