Targeted transcriptional regulation using synthetic transcription factors
AssigneeKWS SAAT SE & Co. KGaA
InventorsAaron Hummel, Mathias Labs
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Abstract
The present invention relates to the targeted regulation of gene expression and more specifically to synthetic transcription factors (STFs) comprising at least one highly target specific engineered recognition domain and further comprising at least one activation or silencing domain to modulate the expression of a gene of interest, preferably to modulate the transcription of a morphogenic gene of a eukaryote. Further disclosed are methods using the STFs to enhance transformation frequencies, to optimize successful genome editing approaches, to provide haploid or double haploid organisms, and/or to provide compositions suitable for general transformation, but also for breeding purposes.
Claims17
A synthetic transcription factor, or a nucleotide sequence encoding the same, comprising at least one recognition domain and at least one activation domain, wherein the synthetic transcription factor is configured to modulate the expression of an endogenous morphogenic gene in a cellular system, wherein the modulation is a targeted activation, and wherein:
- wherein the morphogenic gene comprises a nucleotide sequence selected from the group consisting of:
- (i) a nucleotide sequence set forth in any one of SEQ ID NOs:
- 199, 207, 217, 218, or 219;
- (ii) a nucleotide sequence having the coding sequences of the nucleotide sequence set forth in SEQ ID NOs:
- 199, 207, 217, 218, or 219;
- (iii) the nucleotide sequence complementary to the nucleotide sequence of (i) or (ii);
- (iv) a nucleotide sequence having at least 95% identity to the nucleotide sequence of (i), (ii) or (iii);
- and
- (v) a nucleotide sequence encoding a protein comprising the amino acid sequence set forth in any one of SEQ ID NOs:
- 238 to 240;
- and
- wherein the synthetic transcription factor comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:
- 151 and SEQ ID NO:
- 218, and combinations thereof, and/or the at least one recognition domain comprises a sequence set forth in any one of SEQ ID NOs:
- 13 to 51 or SEQ ID NOs:
- 52 to 94, and
- wherein the synthetic transcription factor and/or the at least one recognition domain binds to a regulation region having the sequence set forth in SEQ ID NOs:
- 95, 109 to 147 or SEQ ID NOs:
- 96, 148 to 190.
A method for increasing the transformation efficiency in a cellular system, wherein the method comprises the steps of:
- (a) providing a cellular system;
- (b) introducing into the cellular system at least one synthetic transcription factor, or a nucleotide sequence encoding the same;
- and
- (c) introducing into the cellular system at least one nucleotide sequence of interest;
- (d) optionally culturing the cellular system under conditions to obtain a transformed progeny of the cellular system;
- wherein the at least one synthetic transcription factor, or the nucleotide sequence encoding the same, comprises at least one recognition domain and at least one activation domain, wherein the synthetic transcription factor is configured to modulate the expression of at least one endogenous morphogenic gene in the cellular system;
- wherein the at least one synthetic transcription factor, or the nucleotide sequence encoding the same, is introduced in parallel to, or sequentially with the introduction of the at least one nucleotide sequence of interest, wherein the modulation is a targeted activation, and wherein:
- wherein the morphogenic gene comprises a nucleotide sequence selected from the group consisting of:
- (i) a nucleotide sequence set forth in any one of SEQ ID NOs:
- 199, 207, 217, 218, or 219;
- (ii) a nucleotide sequence having the coding sequences of the nucleotide sequence set forth in any one of SEQ ID NOs:
- 199, 207, 217, 218, or 219;
- (iii) a nucleotide sequence complementary to the nucleotide sequence of (i) or (ii);
- (iv) a nucleotide sequence having at least 95% identity to the nucleotide sequence of (i), (ii) or (iii);
- and
- (v) a nucleotide sequence encoding a protein comprising the amino acid sequence set forth in any one of SEQ ID NOs:
- 238 to 240;
- wherein the synthetic transcription factor comprises a nucleotide sequence selected from the group consisting of SEQ ID NO:
- 151 and SEQ ID NO:
- 218, and combinations thereof, and/or the at least one recognition domain comprises a sequence set forth in any one of SEQ ID NOs:
- 13 to 51 or SEQ ID NOs:
- 52 to 94, and
- wherein the synthetic transcription factor and/or the at least one recognition domain binds to a regulation region having the sequence set forth in SEQ ID NOs:
- 95, 109 to 147 or SEQ ID NOs:
- 96, 148 to 190.
Dependencies are inferred from the claim text.