# US 11,898,155 B2 · Method for introducing molecule and composition containing inhibitor

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- Number: US 11,898,155 B2
- Document ID: US11898155B2
- Status: Granted
- Kind: B2
- Application number: 16/770,496
- Filed: 2018-12-06
- Published: 2021-06-17
- Granted: 2024-02-13
- Earliest claimed: 2017-12-07
- Assignees: Pearl Kogyo Co., Ltd.; I-Gene Corporation; National University Corporation Ehime University
- Applicants: Pearl Kogyo Co., Ltd.; i-Gene Corporation; National University Corporation Ehime University
- Inventors: Masafumi Jinno; Yuki Isozaki; Yuto Kikuchi; Yugo Kido; Susumu Satoh
- Web page: https://patentgrep.com/patent/US11898155B2

## CPC

- C12N 15/87: Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
- C12N 9/99: Enzyme inactivation by chemical treatment
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

Provided is a method for introducing a selected molecule, including: an inhibitor treatment step of allowing a target cell or a target tissue to internalizing an inhibitor of vesicular transport or lysosomal membrane fusion; and an introduction step of contacting an introduction liquid containing a selected molecule with the target cell or the target tissue to introduce the selected molecule into the target cell or the target tissue.

## Claims

### Claim 1

A method for introducing a molecule, comprising:
allowing a target cell or a target tissue to internalizing an inhibitor of an endocytosis- or autophagy-mediated lysosomal degradation pathway; and
contacting a liquid containing a molecule with the target cell or the target tissue to introduce the molecule into the target cell or the target tissue.

### Claim 2 (depends on 1)

The introduction method according to claim 1, wherein in the contacting, the molecule is introduced via endocytosis into the target cell or the target tissue.

### Claim 3 (depends on 1)

The introduction method according to claim 1, wherein in the contacting, the molecule is introduced by irradiating the target cell or the target tissue with plasma.

### Claim 4 (depends on 1)

The introduction method according to claim 1, wherein the inhibitor comprises a low-molecular-weight compound permeable to a cell membrane of the target cell or a cell as a component of the target tissue.

### Claim 5 (depends on 4)

The introduction method according to claim 4, wherein the low-molecular-weight compound comprises at least one selected from the group consisting of nocodazole, chloroquine, LY-294002, and wortmannin.

### Claim 6 (depends on 1)

The introduction method according to claim 1, wherein in the allowing, the target cell or the target tissue is allowed to internalize the inhibitor by contacting a composition containing the inhibitor with the target cell or the target tissue.

### Claim 7 (depends on 6)

The introduction method according to claim 6, wherein the composition is a culture medium for the target cell or the target tissue.

### Claim 8 (depends on 1)

The introduction method according to claim 1, wherein the molecule comprises a nucleic acid molecule, protein or peptide.

### Claim 9 (depends on 1)

The introduction method according to claim 1, wherein the molecule comprises DNA or RNA.

### Claim 10 (depends on 1)

The introduction method according to claim 1, wherein the molecule comprises a plasmid.

### Claim 11 (depends on 1)

The introduction method according to claim 1, wherein the inhibitor comprises at least one selected from the group consisting of nocodazole, bafilomycin, chloroquine, Lys05, LY-294002 (2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one), wortmannin, 3-methyladenine, cycloheximide, bafilomycin A1, leupeptin, E64d, and pepstatin A.

### Claim 12 (depends on 1)

The introduction method according to claim 1, wherein the introduction liquid comprises water or aqueous solution.

### Claim 13 (depends on 1)

The introduction method according to claim 1, wherein the target cell or target tissue comprises a plurality of target cells.

### Claim 14 (depends on 1)

The introduction method according to claim 1, wherein the target cell or target tissue comprises a prokaryotic cell.

### Claim 15 (depends on 1)

The introduction method according to claim 1, wherein the target cell or target tissue comprises a eukaryotic cell.

### Claim 16 (depends on 1)

The introduction method according to claim 1, wherein the target cell or target tissue comprises a cell from a mouse, a rat, a dog, a rabbit, or a goat.

### Claim 17 (depends on 1)

The introduction method according to claim 1, wherein the target cell or target tissue comprises a donor-derived organ used for transplantation or a pre-differentiation plant tissue constructed by callus culture.