Methods for the Modification of Cells, Modified Cells and Uses Thereof
OwnerLennon James Matchett-Oates, Noel Cogan +2inventors
InventorsLennon James Matchett-Oates, Noel Cogan, German Carlos Spangenberg, Ehab Ibrahim Ahmed Mohamaden
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Abstract
The present disclosure relates generally to methods for the modification of plant cells, e.g., cannabis plant cells, using RNA interference (RNAi), and cells that have been modified to express mediators of RNAi, e.g., short interfering RNA (siRNA). The present disclosure further relates to plants comprising the modified cells and products derived from the plants, e.g., plant material and extracts derived therefrom and methods for selecting siRNA for altering the expression of one or more cannabinoid biosynthesis genes, such as THCAS, CBCAS or CBDAS.
Claims21
1-47. (canceled)
A cannabis cell comprising a targeted genetic modification that alters the expression of one or more cannabinoid biosynthesis genes, wherein the genetic modification is targeted to one or more, or all of:
- a) a region of at least one CBDAS gene comprising a nucleotide sequence corresponding to any one of SEQ ID NOs:
- 18-26, or a or a nucleotide sequence which is at least 90% identical to the nucleotide sequence of SEQ ID NOs:
- 18-26;
- b) a region of at least one CBCAS gene comprising a nucleotide sequence corresponding to any one of SEQ ID NOs:
- 15-17, or a or a nucleotide sequence which is at least 90% identical to the nucleotide sequence of SEQ ID NOs:
- 15-17;
- and
- c) a region of a THCAS gene comprising a nucleotide sequence corresponding to SEQ ID NO:
- 14, or a or a nucleotide sequence which is at least 90% identical to the nucleotide sequence of SEQ ID NO:
- 14,
- and wherein the cell is characterized by altered expression of the one or more cannabinoid biosynthesis genes relative to a cannabis cell that does not comprise the genetic modification.
A synthetic siRNA molecule comprising a nucleotide sequence selected from SEQ ID NOs:
- 27-322, or a nucleotide sequence which is at least 90% identical to the nucleotide sequence of SEQ ID NOs:
- 27-322.
A method for producing a cannabis plant with an altered level or activity, or both, of a cannabinoid biosynthesis pathway, the method comprising:
- a) introducing a targeted genetic modification to a cannabis cell leading to altered expression of one or more cannabinoid biosynthesis genes, wherein the genetic modification is targeted to one or more or all of:
- i) a region of at least one CBDAS gene comprising a nucleotide sequence corresponding to any one of SEQ ID NOs:
- 18-26, or a or a nucleotide sequence which is at least 90% identical to the nucleotide sequence of SEQ ID NOs:
- 18-26;
- ii) a region of at least one CBCAS gene comprising a nucleotide sequence corresponding to any one of SEQ ID NOs:
- 15-17, or a or a nucleotide sequence which is at least 90% identical to the nucleotide sequence of SEQ ID NOs:
- 15-17;
- and
- iii) a region of a THCAS gene comprising a nucleotide sequence corresponding to SEQ ID NO:
- 14, or a or a nucleotide sequence which is at least 90% identical to the nucleotide sequence of SEQ ID NO:
- 14;
- and
- b) regenerating a cannabis plant from the cell of step (a), wherein the plant comprises the genetic modification and has altered level or activity, or both, of the cannabinoid biosynthesis pathway relative to a cannabis plant that does not comprise the genetic modification.
A transgenic cannabis plant comprising a RNA interference (RNAi) vector comprising a nucleotide sequence encoding a siRNA, wherein the siRNA targets one or more or all of a THCAS gene, at least one CBDAS gene and at least one CBCAS gene.
Dependencies are inferred from the claim text.