IP Library › Granted Patent US 12,655,438
Granted Patent B2
US 12,655,438 · App. 17/634,147 · Granted Jun 16, 2026

Augmented sgRNAs and methods for their use to enhance somatic and germline plant genome engineering

Inventors: Daniel F. Voytas (Falcon Heights, MN); Evan Ellison (Elbow Lake, MN)
Assignee: Regents of the University of Minnesota
C12N15/8213C12N9/22C12N15/111C12N15/8203C12N15/825C12N2310/20
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Quick Facts
Patent No.
US 12,655,438
App. No.
17/634,147
Granted
Jun 16, 2026
Kind
B2
Abstract

Methods and materials for increasing somatic and germline genome editing are provided herein. For example, provided herein are methods and materials for using augmented sgRNAs to increase somatic and germline genome editing.

Claims (25)

1 . A method for generating a plant comprising a specific genomic DNA sequence modification, the method comprising:

delivering an augmented sgRNA to a transgenic plant that expresses an RNA guided gene editing reagent, wherein the augmented sgRNA comprises (i) a sequence targeted to the specific genomic DNA sequence and (ii) a mobile RNA sequence; and

recovering, from the plant to which the augmented sgRNA comprising the mobile RNA sequence was delivered, tissue with a genetic modification induced at the specific genomic DNA sequence by the RNA-guided gene editing reagent, wherein the tissue is capable of transmitting the genetic modification to a next generation plant.

2 . The method of claim 1 , wherein the RNA guided gene editing reagent is an RNA guided endonuclease, an RNA guided base editor, or an RNA guided epigenetic modifier.

3 . The method of claim 1 , wherein the mobile RNA sequence is derived from Flowering Time (FT).

4 . The method of claim 1 , wherein the mobile RNA sequence is derived from BEL5, GAI, a tRNA-like motif, or LeT6.

5 . The method of claim 1 , comprising delivering the augmented sgRNA by RNA virus or DNA virus.

6 . The method of claim 1 , comprising delivering the augmented sgRNA by Agrobacterium , biolistics, nanoparticles, or electroporation.

7 . The method of claim 1 ,

wherein the plant gives rise to pollen and egg cells, and wherein the genetic modification is transmitted to the next generation plant, or

wherein the plant comprises edited cells that are regenerated through tissue culture into an edited plant that transmits the genetic modification to the next generation plant.

8 . The method of claim 1 , wherein the sgRNA comprises an RNA sequence that serves as a template for reverse transcription to create a repair template to incorporate a specific sequence change at or near the specific genomic DNA sequence, or wherein the method comprises co-delivering the sgRNA with a DNA that serves as a repair template to incorporate a specific sequence change at or near the specific genomic DNA sequence.

9 . A method for generating a plant comprising a specific genomic DNA sequence modification, the method comprising:

delivering (a) an augmented sgRNA and (b) a sequence encoding an RNA guided gene editing reagent to a plant, wherein the augmented sgRNA comprises (i) a sequence targeted to the specific genomic DNA sequence and (ii) a first mobile RNA sequence, and wherein the sequence encoding the RNA guided gene editing reagent comprises a second mobile RNA sequence; and

recovering, from the plant to which the augmented sgRNA comprising the mobile RNA sequence was delivered, tissue with a genetic modification induced at the specific genomic DNA sequence by the RNA-guided gene editing reagent, wherein the tissue is capable of transmitting the genetic modification to a next generation plant.

10 . The method of claim 9 , wherein the RNA guided gene editing reagent is an RNA guided endonuclease.

11 . The method of claim 9 , wherein the RNA guided gene editing reagent is an RNA guided base editor, an RNA guided epigenetic modifier, or an RNA guided reverse transcriptase.

12 . The method of claim 9 , wherein the first mobile RNA sequence, the second mobile RNA sequence, or both mobile RNA sequences are derived from FT.

13 . The method of claim 9 , wherein the augmented sgRNA comprises a sequence derived from BEL5, GAI, tRNA-like motif, or LeT 6 .

14 . The method of claim 9 , comprising delivering the augmented sgRNA by RNA virus or by DNA virus.

15 . The method of claim 9 , comprising delivering the augmented sgRNA by Agrobacterium , biolistics, nanoparticles, or electroporation.

16 . The method of claim 9 ,

wherein the plant gives rise to pollen and egg cells, and wherein the genetic modification is transmitted to the next generation plant, or

wherein the plant comprises edited cells that are regenerated through tissue culture into an edited plant that transmits the genetic modification to the next generation plant.

17 . The method of claim 9 , wherein the sgRNA comprises an RNA sequence that serves as a repair template to incorporate a specific sequence change at or near the specific genomic DNA sequence, or wherein the sgRNA comprises an RNA sequence that serves as a template for reverse transcription to create a repair template to incorporate a specific sequence change at or near the specific genomic DNA sequence, or wherein the method comprises co- delivering the sgRNA with a DNA that serves as a repair template to incorporate a specific sequence change at or near the specific genomic DNA sequence.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 28, 2022
From: VOYTAS, DANIEL F.; ELLISON, EVAN
To: REGENTS OF THE UNIVERSITY OF MINNESOTA
Reel/Frame 059757/0733 →
Continuity (2)
Provisional Application 62885009 · Aug 9, 2019
Related Publication 20220315938A1 · Oct 6, 2022
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