Systems, methods and composition of using RNase III mutants to produce sRNA to control host pathogen infection
AssigneePebble Labs Inc.
InventorsGuohua Yin, Erick Scott Lebrun, Timothy S. Travers
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Abstract
The current invention includes systems, methods and compositions for the generation of sRNA molecules using select RNase III mutants. In one preferred embodiment, invention includes systems, methods and compositions for the generation of sRNA molecules using RNase III mutants to control a host pathogen through the production and diffusion of sRNA molecules that may initiate an RNAi pathway response directed to a host pathogen. Additional embodiments of the current invention include systems, methods and compositions for the DICER-independent generation of sRNA molecules using select RNase III mutants.
Claims20
A genetically modified cell expressing a heterologous polynucleotide sequence operably linked to a promoter sequence encoding an RNase III mutant from E. coli or Enterobacter adapted for enhanced generation of small RNA (sRNA) from catalytic cutting of double stranded RNA (dsRNA), wherein said RNase III mutant comprises:
- an E38A-R107A-R108A RNase III mutant, wherein a glutamic acid is replaced with an alanine at residue 38, and an arginine is replaced with an alanine at residue 107, and an arginine is replaced with an alanine at residue 108, or a homologous RNase III mutant thereof, wherein the homologous RNase III's wild-type sequence includes the conserved residues E38, R107 and R108.
A composition comprising an E38A-R107A-R108A RNase III mutant from E. coli or Enterobacter, wherein a glutamic acid is replaced with an alanine at residue 38, and an arginine is replaced with an alanine at residue 107, and an arginine is replaced with an alanine at residue 108.
A composition comprising polynucleotide sequence operably linked to a promoter sequence encoding an E38A-R107A-R108A RNase III mutant selected from:
- an E38A-R107A-R108A RNase III mutant according to SEQ ID NO. 7, or a fragment thereof;
- an E38A-R107A-R108A RNase III mutant according to SEQ ID NO. 11, or a fragment thereof;
- an E38A-R107A-R108A RNase III mutant according to SEQ ID NO. 15, or a fragment thereof.
A composition comprising an E38A-R107A-R108A RNase III mutant polypeptide mutant selected from:
- an E38A-R107A-R108A RNase III mutant polypeptide according to SEQ ID NO. 12, or a fragment thereof;
- an E38A-R107A-R108A RNase III mutant polypeptide according to SEQ ID NO. 16, or a fragment thereof;
- an E38A-R107A-R108A RNase III mutant polypeptide according to SEQ ID NO. 8 or a fragment thereof.
Dependencies are inferred from the claim text.