Antimicrobial compositions and methods of use thereof
Antimicrobial compositions including a cell penetrating peptide (CPP) having the amino acid sequence Tyr-Ala-Arg-Val-Arg-Arg-Arg-Gly-Pro-Arg-Gly-Tyr-Ala-Arg-Val-Arg-Arg-Arg-Gly-Pro-Arg-Arg (SEQ ID NO:1) or variant thereof are disclosed. The CPP, which itself has antimicrobial properties, can be advantageously combined with or conjugated to a cargo to increase the delivery, efficacy, or combinations thereof, of the cargo into cells. In preferred embodiments, the CPP is combined with or conjugated to a functional nucleic acid, such as an external guide sequence (EGS) which can target and reduce expression of essential microbial genes or genes than impart resistance to antimicrobial drugs. Methods of using the compositions alone or in combination with traditional antimicrobial drugs to treat infections are also disclosed.
1. A composition comprising
a basic cell penetrating peptide (CPP) comprising the amino acid sequence of SEQ ID NO:1 or a variant of SEQ ID NO:1 having at least 90% sequence identity to SEQ ID NO:1,
and an external guide sequence (EGS).
2. The composition of claim 1 wherein the EGS is conjugated to the CPP.
3. The composition of claim 2 wherein the EGS is covalently conjugated to the CPP.
4. The composition of claim 1 wherein the EGS comprises a sequence antisense to a target mRNA.
5. The composition of claim 4 wherein the target mRNA is encoded by an essential microbial gene or antimicrobial resistance gene.
6. The composition of claim 4 wherein the target mRNA is encoded by a gene comprising the nucleic acid sequence SEQ ID NO: 4, 5, 6, 7, 8, 9, 12, 15, 16, 17, 18, or 19.
7. The composition of claim 1 wherein the EGS comprises the nucleic acid sequence SEQ ID NO: 14, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, or 37.
8. The composition of claim 1 wherein the EGS is a morpholino oligonucleotide.
9. The composition of claim 1 wherein the composition further comprises an antimicrobial drug.
10. The composition of claim 1 wherein the CPP is not a fusion partner in a fusion protein.
11. The composition of claim 1 wherein the CPP comprises the amino acid sequence of SEQ ID NO: 1, 2, 3, 49, 50, 51, 52, or 53.
12. The composition of claim 1 , wherein the CPP is conjugated to the EGS, and wherein the EGS comprises a sequence comprising five nucleotides of complementarity to an mRNA encoding an essential microbial gene or antimicrobial resistance gene.
13. The composition of claim 12 , wherein the CPP-EGS conjugate binds more tightly to the mRNA relative to an unconjugated EGS having the same sequence.
14. The composition of claim 12 , wherein the CPP comprises the amino acid sequence of SEQ ID NO:1.