# US 8,785,612 B2 · Sugarcane bacilliform viral (SCBV) enhancer and its use in plant functional genomics

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- Number: US 8,785,612 B2
- Document ID: US8785612B2
- Status: Granted
- Kind: B2
- Application number: 13/220,564
- Filed: 2011-08-29
- Published: 2012-03-01
- Granted: 2014-07-22
- Earliest claimed: 2010-08-30
- Assignees: Agrigenetics, Inc.
- Applicants: John P. Davies; Vaka S. Reddy; William M. Ainley; Mark A. Thompson
- Inventors: John P. Davies; Vaka S. Reddy; William M. Ainley; Mark A. Thompson
- Web page: https://patentgrep.com/patent/US8785612B2

## CPC

- C12N 15/8216: {Methods for controlling, regulating or enhancing expression of transgenes in plant cells}
- 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)}
- C12N 15/8241: {Phenotypically and genetically modified plants via recombinant DNA technology}
- C12N 15/8261: {with agronomic (input) traits, e.g. crop yield}
- Y02A 40/146: Genetically Modified [GMO] plants, e.g. transgenic plants
- C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00)
- Y02A: TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE

## Abstract

Identification of new enhancer sequence has significant utility in the plant functional genomics. The sugarcane bacilliform badnavirus (SCBV) transcriptional enhancer has been identified. This enhancer can be used to increase the rate of transcription from gene promoters and in activation tagging experiments. A ten-fold increase in transcription was observed when a 4× array of the SCBV enhancer was placed upstream of a truncated form of the maize alcohol dehydrogenase minimal promoter. Methods of using the SCBV transcriptional enhancer are described, as are chimeric transcription regulatory regions, constructs, cells, tissues, and organisms that comprise one or more copies of the enhancer.

## Claims

### Claim 1

A chimeric transcription regulatory region comprising:
a sugarcane bacilliform viral (SCBV) enhancer element consisting of position 337 to position 618 of SEQ ID NO: 1; and
operably linked thereto, a heterologous promoter comprising an RNA polymerase binding site and a mRNA initiation site,
wherein the chimeric transcription regulatory region enhances transcription of an operably linked nucleotide sequence of interest, compared to transcription of the nucleotide sequence of interest operably linked to a chimeric transcription regulatory region comprising the promoter and not comprising the SCBV enhancer sequence.

### Claim 2 (depends on 1)

The chimeric transcription regulatory region of claim 1, further comprising two or more copies of the sugarcane bacilliform viral (SCBV) enhancer element consisting of position 337 to position 618 of SEQ ID NO: 1.

### Claim 3 (depends on 1)

The chimeric transcription regulatory region of claim 1, wherein the promoter is obtained from the upstream region of a plant virus gene, a bacterial gene, a fungal gene, a plant nuclear gene, a plant extra-nuclear gene, an invertebrate gene, or a vertebrate gene.

### Claim 4 (depends on 1)

A construct comprising the transcription regulatory region of claim 1 operably linked to a transcribable polynucleotide molecule operably linked to a 3′ transcription termination polynucleotide molecule.

### Claim 5 (depends on 4)

The construct of claim 4, wherein said transcribable polynucleotide molecule confers an agronomic trait to a plant in which it is expressed.

### Claim 6 (depends on 4)

A transgenic plant stably transformed with the construct of claim 4.

### Claim 7 (depends on 6)

The transgenic plant of claim 6, wherein the transcribable polynucleotide molecule confers an agronomic trait to a plant in which it is expressed.

### Claim 8 (depends on 6)

A seed of the transgenic plant of claim 6, wherein the seed comprises the construct.

### Claim 9 (depends on 6)

The transgenic plant of claim 6, wherein the plant is a maize plant.

### Claim 10 (depends on 1)

A transgenic plant cell comprising the chimeric transcription regulatory region of claim 1.

### Claim 11 (depends on 4)

A method of producing a transgenic plant comprising:
(i) transforming a plant cell or tissue with the construct of claim 4; and
(ii) regenerating the transgenic plant from the transformed plant cell or tissue, thereby producing a transgenic plant.

### Claim 12 (depends on 11)

The method of claim 11, wherein the transgenic plant is a dicotyledon.

### Claim 13 (depends on 11)

The method of claim 11, wherein the transgenic plant is a monocotyledon.

### Claim 14 (depends on 4)

A plant cell or tissue transformed with the construct of claim 4.

### Claim 15 (depends on 14)

The plant cell or tissue of claim 14, wherein the plant cell or tissue is from a dicotyledon.

### Claim 16 (depends on 14)

The plant cell or tissue of claim 14, wherein the plant cell or tissue is derived from a monocotyledon.

### Claim 17 (depends on 5)

A transgenic plant cell, fruit, leaf, root, shoot, flower, seed, cutting or other reproductive material useful in sexual or asexual propagation, transgenic progeny plant inclusive of an F1 hybrid, transgenic male-sterile plant or any other transgenic plant or transgenic plant product, derivable from the transgenic plant of claim 5, wherein the transgenic plant cell, fruit, leaf, root, shoot, flower, seed, cutting or other reproductive material useful in sexual or asexual propagation, transgenic male-sterile plant, or other transgenic plant, or transgenic plant product comprises the construct.

### Claim 18

A maize plant cell, tissue or plant comprising two or more copies of the sugarcane bacilliform viral (SCBV) enhancer element consisting of position 337 to position 618 of SEQ ID NO: 1, in which the two or more copies of the SCBV enhancer element is inserted into a genome of the maize plant cell, tissue or plant at a random location.

### Claim 19 (depends on 18)

The maize plant cell, tissue or plant of claim 18, wherein the SCBV enhancer imparts enhanced transcription of a nucleotide sequence of interest which is under regulatory control of the SCBV enhancer and a promoter, as compared to transcription of the nucleotide sequence of interest in the absence of the SCBV enhancer.

### Claim 20 (depends on 2)

The chimeric transcription regulatory region of claim 2, comprising at least three copies of the SCBV enhancer element.

### Claim 21 (depends on 2)

The chimeric transcription regulatory region of claim 2, comprising at least four copies of the SCBV enhancer element.