Chimeric proteins for targeting dsRNA
Described herein are recombinant chimeric proteins comprising a double stranded RNA (dsRNA) binding domain and a cancer-cell targeting domain for targeting of dsRNA to cancer cells. Methods of use of the described chimeric proteins are also provided herein.
1. A recombinant protein comprising a double stranded RNA (dsRNA) binding domain and a target-binding moiety, wherein said target-binding moiety is EGF and said dsRNA binding domain comprises one or more double-stranded RNA-binding motifs (dsRBM), wherein at least one of said one or more dsRBMs is a dsRBM of dsRNA dependent protein kinase R (PKR), and wherein said recombinant protein further comprises a linker between the dsRNA binding domain and the target-binding moiety, wherein said linker is a peptide of sequence ACSGSACSGSAGNRVRRSVGSSNG (SEQ ID NO: 26).
2. The recombinant protein of claim 1 , wherein said dsRNA binding domain comprises two dsRBMs of a PKR, optionally connected by a flexible linker.
3. The recombinant protein of claim 2 , wherein said dsRNA binding domain is selected from the dsRNA binding domain of human PKR and said two dsRBMs are connected by a flexible linker; or the full length human PKR.
4. The recombinant protein of claim 3 , wherein said dsRNA binding domain comprises amino acid residues 1-197 of human PKR.
5. The recombinant protein of claim 1 , wherein said EGF is human EGF.
6. A complex comprising the recombinant protein of claim 1 and dsRNA.
7. The complex of claim 6 , wherein said dsRNA is dsRNA comprising a polyinosinic acid strand and a polycytidylic acid strand (poly IC).
8. The recombinant protein of claim 1 , wherein said dsRNA binding domain comprises two dsRBMs of human PKR connected by a flexible linker.
9. The recombinant protein of claim 1 , wherein said dsRNA binding domain comprises the amino acid sequence of SEQ ID NO:18.
10. The recombinant protein of claim 1 , wherein said dsRNA binding domain consists of the amino acid sequence of SEQ ID NO:18.
11. The recombinant protein of claim 2 , wherein said EGF is human EGF.
12. The recombinant protein of claim 8 , wherein said EGF is human EGF.
13. The recombinant protein of claim 9 , wherein said EGF is human EGF.
14. The recombinant protein of claim 10 , wherein said EGF is human EGF.
15. A complex comprising the recombinant protein of claim 6 and dsRNA.
16. The complex of claim 15 , wherein said dsRNA is dsRNA comprising a polyinosinic acid strand and a polycytidylic acid strand (poly IC).
17. A complex comprising the recombinant protein of claim 13 and dsRNA.
18. The complex of claim 17 , wherein said dsRNA is dsRNA comprising a polyinosinic acid strand and a polycytidylic acid strand (poly IC).
19. A complex comprising the recombinant protein of claim 14 and dsRNA.
20. The complex of claim 19 , wherein said dsRNA is dsRNA comprising a polyinosinic acid strand and a polycytidylic acid strand (poly IC).