US 11,999,958 B2
Methods of gene editing and transforming Cannabis
Michael W. Petersen, Merrimac, WI (US); Edward James Williams, Madison, WI (US); Robert Harnish, Middleton, WI (US); Heidi Flewelling Kaeppler, Oregon, WI (US); Brian Martinell, Mt. Horeb, WI (US); Ray Collier, Middleton, WI (US); Frank McFarland, Madison, WI (US); and Shawn Michael Kaeppler, Oregon, WI (US)
Assigned to WISCONSIN ALUMNI RESEARCH FOUNDATION, Madison, WI (US)
Filed by Wisconsin Alumni Research Foundation, Madison, WI (US)
Filed on Jul. 16, 2020, as Appl. No. 16/930,936.
Claims priority of provisional application 62/982,522, filed on Feb. 27, 2020.
Claims priority of provisional application 62/906,210, filed on Sep. 26, 2019.
Claims priority of provisional application 62/875,311, filed on Jul. 17, 2019.
Prior Publication US 2021/0071186 A1, Mar. 11, 2021
Int. Cl. C12N 15/82 (2006.01); A01H 4/00 (2006.01); A01H 6/28 (2018.01); C07K 14/415 (2006.01)
CPC C12N 15/8201 (2013.01) [A01H 4/005 (2013.01); A01H 6/28 (2018.05); C07K 14/415 (2013.01)] 11 Claims
 
1. A method of transforming a Cannabis plant, the method comprising:
growing a sanitized and imbibed Cannabis seed on a non-selective culture medium suitable for supporting the growth and survival of the Cannabis seed;
harvesting an internode, leaf, or petiole tissue to form a Cannabis explant;
inoculating the Cannabis explant with a heterologous nucleic acid;
co-culturing the Cannabis explant in a co-culture medium for between about 1 day and about 6 days,
culturing the Cannabis explant on a selection medium comprising meta-topolin to select for transformed Cannabis explants.