# US 11,299,733 B2 · Process of delivering small RNAs to sperm

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 11,299,733 B2
- Document ID: US11299733B2
- Status: Granted
- Kind: B2
- Application number: 16/315,004
- Filed: 2017-07-12
- Published: 2019-10-10
- Granted: 2022-04-12
- Earliest claimed: 2016-07-15
- Assignees: University of Massachusetts
- Applicants: University of Massachusetts
- Inventors: Oliver Rando; Upasna Sharma; Colin Conine
- Web page: https://patentgrep.com/patent/US11299733B2

## CPC

- C12N 15/113: Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; {Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing (when used in plants C12N15/8218)}
- A61B 17/43: for artificial insemination
- A61B 17/435: for embryo {or ova} transplantation
- A61D 19/02: for artificial insemination
- A61D 19/04: for embryo transplantation
- A61K 9/0034: {Urogenital system, e.g. vagina, uterus, cervix, penis, scrotum, urethra, bladder; Personal lubricants}
- A61K 9/06: Ointments; Bases therefor; {Other semi-solid forms, e.g. creams, sticks, gels (composition of ointments, creams or gels A61K47/00)}
- C12N 5/0604: {Whole embryos; Culture medium therefor}
- C12N 5/061: {Sperm cells, spermatogonia}
- C12N 15/111: {General methods applicable to biologically active non-coding nucleic acids}
- C12N 2310/14: interfering nucleic acids [NA]
- C12N 2310/141: MicroRNAs, miRNAs
- C12N 2320/30: Special therapeutic applications
- C12N 2501/65: MicroRNA
- C12N 2510/00: Genetically modified cells
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- A61B: DIAGNOSIS; SURGERY; IDENTIFICATION (usefulness limited to only animals A61D)
- A61D: VETERINARY INSTRUMENTS, IMPLEMENTS, TOOLS, OR METHODS
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)

## Abstract

Methods and compositions directed to altering a population of sRNAs in a sperm using vesicles isolated from an epididymosome are provided. Methods and compositions directed to altering a population of sRNAs in an oocyte using vesicles isolated from an epididymosome are also provided. Methods for altering an sRNA population in a sperm or an oocyte can be used to prevent, or reduce the severity of, a disease, disorder, or condition that would otherwise be inherited by progeny. For example, certain epigenetic inherited conditions due to paternal effects, such as certain metabolic and stress disorders and conditions, can be ameliorated in progeny using sperm or oocytes having an altered sRNA population.

## Claims

### Claim 1

A method of altering the sRNA population in a sperm, comprising contacting the sperm with a vesicle comprising an epididymosome loaded with a molecular cargo comprising a synthetic sRNA to produce a sperm having an altered sRNA population, wherein the sperm having an altered sRNA population is capable of fertilizing an oocyte.

### Claim 2 (depends on 1)

The method of claim 1, wherein the sRNA is selected from the group consisting of a siRNA, a miRNA, a piRNA, a snoRNA, a srRNA, a U-RNA, and a tRNA fragment.

### Claim 3 (depends on 1)

The method of claim 1, wherein the sRNA is a tRNA fragment selected from the group consisting of a tRNA-Gly-CCC fragment, a tRNA-Gly-TCC fragment, a tRNA-Gly-GCC fragment, a tRNA-Val-CAC fragment, a tRNA-Glu-CTC fragment, a tRNA-Lys-CTT fragment, and a tRNA-His-GTG fragment.

### Claim 4 (depends on 1)

The method of claim 1, wherein the epididymosome is selected from the group consisting of caput epididymosome, corpus epididymosome and cauda epididymosome.

### Claim 5 (depends on 1)

The method of claim 1, wherein the resulting sperm comprises an sRNA population comprising increased siRNA, miRNA, piRNA, snoRNA, srRNA, U-RNA, or tRNA fragment content.

### Claim 6 (depends on 5)

The method of claim 5, wherein the tRNA fragment content comprises a tRNA-Gly-CCC fragment content, tRNA-Gly-TCC fragment content, tRNA-Gly-GCC fragment content, tRNA-Val-CAC fragment content, tRNA-Glu-CTC fragment content, tRNA-Lys-CTT fragment content, or tRNA-His-GTG fragment content.

### Claim 7 (depends on 5)

The method of claim 5, wherein the miRNA is selected from the group consisting of miR-10a/b, miR-141, miR-143, miR-148 and miR-200a.

### Claim 8 (depends on 1)

The method of claim 1, wherein the vesicle comprises molecular cargo comprising a transgene.

### Claim 9 (depends on 1)

The method of claim 1, wherein the sperm is capable of fertilizing an oocyte in vitro.

### Claim 10 (depends on 1)

The method of claim 1, wherein the sperm is capable of fertilizing an oocyte in vivo.

### Claim 11 (depends on 1)

The method of claim 1, wherein prior to contacting the sperm, the sperm is frozen.

### Claim 12 (depends on 1)

The method of claim 1, wherein the vesicle is loaded with molecular cargo using electroporation.

### Claim 13 (depends on 1)

The method of claim 1, wherein the vesicle is loaded with molecular cargo using co-incubation.