# US 8,148,144 B2 · pCryptoRNAi

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- Number: US 8,148,144 B2
- Document ID: US8148144B2
- Status: Granted
- Kind: B2
- Application number: 12/410,748
- Filed: 2009-03-25
- Published: 2009-10-15
- Granted: 2012-04-03
- Earliest claimed: 2008-03-27
- Assignees: National University Corporation Chiba University
- Inventors: Jon Ander Ochoa de Eribe Casas; Susumu Kawamoto
- Web page: https://patentgrep.com/patent/US8148144B2

## CPC

- C12N 15/80: for fungi
- C12N 15/111: {General methods applicable to biologically active non-coding nucleic acids}
- C12N 2330/51: Specially adapted vectors
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)

## Abstract

A vector developed to transform fungi can be used to study the expression of a gene of interest. The vector can provide for the expression of signal proteins in fungi that can be observed and/or monitored. The vector can be used to investigate the effects of RNA interference on a gene of interest in pathogenic fungi. Systems and methods of using the vector are provided.

## Claims

### Claim 1

A nucleic acid comprising the nucleic acid SEQ ID NO: 1.

### Claim 2 (depends on 1)

A vector comprising the nucleic acid of claim 1.

### Claim 3 (depends on 2)

The vector of claim 2, wherein the vector is a plasmid vector.

### Claim 4 (depends on 3)

The vector of claim 3, further comprising a gene of interest inserted into a cloning site in the vector.

### Claim 5 (depends on 4)

The vector of claim 4, wherein the gene of interest comprises a double-stranded RNA sequence capable of activating RNA-dependent Induced Silencing Complex (RISC).

### Claim 6 (depends on 3)

A transformed fungus comprising the vector of claim 3.

### Claim 7 (depends on 6)

The transformed fungus of claim 6, wherein the vector is stably incorporated into the genome of the fungus.

### Claim 8 (depends on 3)

A method of transforming a fungus, wherein the vector of claim 3 is introduced into the fungus genome.

### Claim 9

A method of assessing the effects of RNA interference in a fungus, comprising:
transforming the fungus with a plasmid vector comprising SEQ ID NO: 1 and a gene of interest inserted into a cloning site in the plasmid vector; and
detecting the presence or absence of one or more of a first signal protein and a second signal protein encoded by the plasmid vector.

### Claim 10 (depends on 4)

An in vivo assay system for determining the effect of RNA interference on fungi comprising:
a composition of fungi cells;
the vector of claim 4; and
an imaging device.

### Claim 11 (depends on 10)

The in vivo assay system of claim 10, wherein the fungi comprise Cryptococcus.