Patent ID: 11859219
Assignee: FLAGSHIP PIONEERING INNOVATIONS V, INC.
Field: Biotechnology (Chemistry)
Classification: CPC C  A | IPC A  C

Claim 0:
1. A method of altering a target nucleotide sequence in a population of protoplasts of a plant, comprising delivering a preassembled ribonucleoprotein composition into the population of protoplasts, wherein the preassembled ribonucleoprotein composition comprises:
(a) an RNA-guided nuclease protein, and
(b) a guide RNA (gRNA) that has a nucleotide sequence designed to alter a target nucleotide sequence in the population of protoplasts, and is provided as:
(i) a CRISPR RNA (crRNA) that comprises the gRNA, or
(ii) a single guide RNA (sgRNA) that comprises the gRNA;

wherein the RNA-guided nuclease protein of (a) is complexed with the gRNA of (b); and
wherein the delivering comprises contacting the population of protoplasts with the preassembled ribonucleoprotein composition and concurrently incubating the population of protoplasts at 37 degrees Celsius for a period of between 10 minutes to about 2 hours, thereby altering the target nucleotide sequence.