Methods and Compositions for Selective Regulation of Protein Expression

Patent metadata

Field	Value
Publication number	US-2016208283-A1
Application number	US-201514720052-A
Country	US
Kind code	A1
Filing date	May 22, 2015
Priority date	Jul 1, 2011
Publication date	Jul 21, 2016
Grant date	—

Abstract

The invention provides methods and compositions for selectively suppressing the expression of a recombinant protein in a male reproductive tissue of a transgenic plant. The invention also provides methods and compositions for inducing male sterility in a transgenic plant. Plants, plant cells, plant parts, seeds, and commodity products including such compositions are aspects of the invention.

First claim

1 . A recombinant DNA construct comprising a protein-coding sequence operably linked to a DNA sequence comprising a male tissue-specific siRNA (mts-siRNA) element, wherein said mts-siRNA element is heterologous with respect to said protein-coding sequence.

2 . The recombinant DNA construct of claim 1 , wherein said mts-siRNA element comprises at least one mts-siRNA sequence.

3 . The recombinant DNA construct of claim 2 , wherein said mts-siRNA sequence comprises any one or more of SEQ ID NO: 1-56 and 105-149.

4 . The recombinant DNA construct of claim 1 , wherein said mts-siRNA element comprises any one or more of SEQ ID NO: 57-86, 92-94 and 96-104.

5 . The recombinant DNA construct of claim 1 , wherein the expression of said protein-coding sequence in a transgenic plant confers at least vegetative herbicide tolerance to said plant.

6 . The recombinant DNA construct of claim 5 , wherein said protein-coding sequence encodes a glyphosate-tolerant EPSPS.

7 . A method of making a recombinant DNA construct comprising
a) identifying an mts-siRNA element comprising at least one mts-siRNA sequence, and b) operably linking said mts-siRNA element to a protein-coding sequence, wherein said mts-siRNA element is heterologous with respect to said protein-coding sequence.

8 . The method of claim 7 , wherein said at least one mts-siRNA sequence comprises any one or more of SEQ ID NO: 1-56 and 105-149 or said mts-siRNA element comprises any one or more of SEQ ID NO: 57-86, 92-94 and 96-104.

9 . The method of claim 7 , wherein said mts-siRNA element is tassel-specific.

10 . A transgenic plant having in its genome the recombinant DNA construct of claim 1 .

11 . A seed, progeny, or plant part of the transgenic plant of claim 10 .

12 . The transgenic plant of claim 10 , wherein said transgenic plant is a monocotyledonous plant.

13 . The transgenic plant of claim 10 , wherein said transgenic plant is a maize plant.

14 . A method of selectively suppressing the expression of a recombinant protein in a male reproductive tissue of a transgenic plant comprising expressing in said transgenic plant a recombinant DNA construct of claim 1 .

15 . (canceled)

16 . (canceled)

17 . The method of claim 14 , wherein said mts-siRNA element comprises at least one mts-siRNA sequence comprising any one or more of SEQ ID NO: 1-56 and 105-149.

18 . The method of claim 14 , wherein said wherein said mts-siRNA element comprises any one or more of SEQ ID NO: 57-86, 92-94 and 96-104.

19 . (canceled)

20 . (canceled)

21 . A method of inducing male-sterility in a transgenic plant comprising applying an effective amount of an herbicide to a transgenic plant comprising a recombinant DNA construct of claim 1 , wherein said herbicide application is carried out during the development of the male reproductive tissue of said transgenic plant thereby inducing male-sterility in said transgenic plant.

22 . (canceled)

23 . (canceled)

24 . (canceled)

25 . (canceled)

26 . (canceled)

27 . (canceled)

28 . (canceled)

29 . (canceled)

30 . (canceled)

31 . Hybrid seed harvested from a male-sterile transgenic plant that has been fertilized with pollen from a second plant, wherein said male-sterile transgenic plant comprises a recombinant DNA construct of comprising a protein-coding sequence operably linked to a DNA sequence comprising a male tissue-specific siRNA (mts-siRNA) element, wherein said mts-siRNA element is heterologous with respect to said protein-coding sequence and has been induced to be male-sterile by application of an effective amount of herbicide during the development of the male reproductive tissue of said transgenic plant.

32 . The transgenic plant of claim 10 , wherein said mts-siRNA element is recognized by at least one mts-siRNA produced from a dsRNA formed between transcripts of opposite polarity.

33 . The transgenic plant of claim 10 , wherein said mts-siRNA element is recognized by at least one mts-siRNA, the production of which requires RNA-dependent RNA polymerase activity.

Assignees

• Monsanto Technology Llc

Inventors

• Huang Jintai
• Ivashuta Sergey
• Qi Youlin
• Wiggins Barbara Elizabeth
• Zhang Yuanji

Classifications

• C12N15/8289 Primary

  Male sterility · CPC title

• C12N15/8218

  Antisense, co-suppression, viral induced gene silencing [VIGS], post-transcriptional induced gene silencing [PTGS] · CPC title

• A01N37/46

  N-acyl derivatives · CPC title

• C12N15/8275

  Glyphosate · CPC title

• A01N65/44

  Poaceae or Gramineae [Grass family], e.g. bamboo, lemon grass or citronella grass · CPC title