Methods and compositions for generating dominant short stature alleles using genome editing

A method for producing a mutant allele of the endogenous GA20 oxidase_3 locus, the method comprising:

• a. generating a double-stranded break in the endogenous GA20 oxidase_3 locus in a corn cell using a targeted editing technique;
• and
• b. inserting at the double-stranded break a DNA segment, wherein the DNA segment encodes an antisense RNA sequence that is at least 70% complementary to at least 20, consecutive nucleotides of one or more of SEQ ID NOs:
• 1-3 and 5-7, and wherein the mutant allele of the endogenous GA20 oxidase_3 locus produces a RNA transcript comprising the antisense RNA sequence.

A method for producing a mutant allele of the endogenous GA20 oxidase_3 locus, the method comprising:

• a. generating a first double-stranded break and a second double strand break in the endogenous GA20 oxidase_3 locus in a corn cell using a targeted editing technique;
• and
• b. inserting a DNA segment between the first double-stranded break and the second double-stranded break, wherein the DNA segment encodes an antisense RNA sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% complementary to at least 20, at least 25, at least 30, at least 40, at least 50, at least 75, at least 100, at least 150, at least 200, at least 300, at least 400, at least 500, at least 750, or at least 1000 consecutive nucleotides of one or more of SEQ ID NOs:
• 1-3 and 5-7, and wherein the mutant allele of the endogenous GA20 oxidase_3 locus produces a RNA transcript comprising the antisense RNA sequence.

A method for producing a mutant allele of the endogenous GA20 oxidase_5 locus, the method comprising:

• a. generating a double-stranded break (DSB) in the endogenous GA20 oxidase_5 locus in a corn cell using a targeted editing technique;
• and
• b. inserting at the DSB a DNA segment using a targeted editing technique, wherein the DNA segment encodes an antisense RNA sequence that is at least 70% complementary to at least 20 consecutive nucleotides of one or more of SEQ ID NOs:
• 1-3 and 5-7, and wherein the mutant allele of the endogenous GA20 oxidase_5 locus produces a RNA transcript comprising the antisense RNA sequence.

A method for producing a mutant allele of the endogenous GA20 oxidase_5 locus, the method comprising:

• a. generating a first double-stranded break and a second double strand break in the endogenous GA20 oxidase_5 locus in a corn cell using a targeted editing technique;
• and
• b. inserting a DNA segment between the first double-stranded break and the second double-stranded break, wherein the DNA segment encodes an antisense RNA sequence that is at least 70%, at least 75%, at least 80%, at least 85%, at least 90%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, or 100% complementary to at least 20, at least 25, at least 30, at least 40, at least 50, at least 75, at least 100, at least 150, at least 200, at least 300, at least 400, at least 500, at least 750, or at least 1000 consecutive nucleotides of one or more of SEQ ID NOs:
• 1-3 and 5-7, and wherein the mutant allele of the endogenous GA20 oxidase_5 locus produces a RNA transcript comprising the antisense RNA sequence.

A method for generating a corn plant comprising:

• (a) fertilizing at least one female corn plant with pollen from a male corn plant, wherein the at least one female corn plant and/or the male corn plant comprise(s) a mutant allele of an endogenous GA20 oxidase locus, wherein the mutant allele comprises a DNA segment inserted into the endogenous GA20 oxidase locus, wherein the DNA segment encodes an antisense RNA sequence that is at least 70% complementary to at least 20 consecutive nucleotides of one or more of SEQ ID NOs:
• 1-3 or 5-7, and wherein the mutant allele of the endogenous GA20 oxidase locus produces a RNA transcript comprising the antisense RNA sequence;
• and
• (b) obtaining at least one seed produced by said fertilizing of step (a).