US 12,344,853 B2
bZIP transcription factors regulate conversion of nicotine to nornicotine and reduce levels of tobacco specific (TSNA) precursors
Ling Yuan, Lexington, KY (US); Sanjay K. Singh, Lexington, KY (US); Sitakanta Pattanaik, Lexington, KY (US); and Darlene Madeline Lawson, Chapel Hill, NC (US)
Assigned to University of Kentucky Research Foundation, Lexington, KY (US); and R. J. Reynolds Tobacco Company, Winston-Salem, NC (US)
Appl. No. 17/605,506
Filed by University of Kentucky Research Foundation, Lexington, KY (US); and R.J. Reynolds Tobacco Company, Winston-Salem, NC (US)
PCT Filed Jun. 5, 2020, PCT No. PCT/US2020/036426
§ 371(c)(1), (2) Date Oct. 21, 2021,
PCT Pub. No. WO2020/247821, PCT Pub. Date Dec. 10, 2020.
Claims priority of provisional application 62/857,718, filed on Jun. 5, 2019.
Prior Publication US 2022/0213496 A1, Jul. 7, 2022
This patent is subject to a terminal disclaimer.
Int. Cl. C12N 15/82 (2006.01); C07K 14/415 (2006.01); A24B 3/08 (2006.01)
CPC C12N 15/8243 (2013.01) [C07K 14/415 (2013.01); C12N 15/8218 (2013.01); A24B 3/08 (2013.01)] 40 Claims
 
1. A method of decreasing conversion of nicotine to nornicotine and/or a method of reducing levels of at least one tobacco specific (TSNA) precursor, the method comprising
reducing the activity and/or expression of at least one basic region/leucine zipper (bZIP) type transcription factor in a tobacco plant, a part of a tobacco plant, a tobacco plant cell or cell culture, or a tobacco plant seed, wherein the bZIP transcription factor comprises
an amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 1 or 3;
an amino acid having the sequence of SEQ ID NO: 2, 4, 5, or 6;
an amino acid sequence having at least 95% identity to the amino acid sequence encoded by the nucleic acid sequence of SEQ ID NO: 1 or 3 and which binds a promoter of a nicotine N-demethylase (NND) selected from the group consisting of CYP82E4v1, CYP82E5v2, and CYP82E10;
an amino acid sequence having at least 95% identity to the amino acid sequence of SEQ ID NO: 2, 4, 5, or 6 and which binds a promoter of a nicotine N-demethylase (NND) selected from the group consisting of CYP82E4v1, CYP82E5v2, and CYP82E10; or
combinations thereof.