INIR4 TRANSGENIC MAIZE
Transgenic INIR4 maize plants comprising modifications of the DAS59122-7 maize locus which provide for facile excision of the modified DAS59122-7 transgenic locus or portions thereof, methods of making such plants, and use of such plants to facilitate breeding are disclosed.
1 . A transgenic maize plant cell comprising an INIR4 transgenic locus comprising an originator guide RNA recognition site (OgRRS) in a first DNA junction polynucleotide of a DAS59122-7 transgenic locus and a cognate guide RNA recognition site (CgRRS) in a second DNA junction polynucleotide of the DAS59122-7 transgenic locus.
2 . A transgenic maize plant cell comprising an INIR4 transgenic locus comprising an insertion and/or substitution of DNA in a DNA junction polynucleotide of a DAS59122-7 transgenic locus with DNA comprising a cognate guide RNA recognition site (CgRRS) or a deletion in a 5′ or 3′ junction polynucleotide of a DAS59122-7 transgenic locus.
3 . The transgenic maize plant cell of claim 1 , wherein said CgRRS comprises the DNA molecule set forth in SEQ ID NO: 8, 9, or 10.
4 . The transgenic maize plant cell of claim 1 , wherein said DAS59122-7 transgenic locus is set forth in SEQ ID NO:1, is present in seed deposited at the ATCC under accession No. PTA-11384 is present in progeny thereof, is present in allelic variants thereof, or is present in other variants thereof.
5 . The transgenic maize plant cell of claim 1 , wherein said INIR4 transgenic locus comprises the DNA molecule set forth in SEQ ID NO: 17, 3, 27, or an allelic variant thereof.
6 . The transgenic maize plant cell of claim 2 , wherein said INIR4 transgenic locus comprises the DNA molecule set forth in SEQ ID NO: 2, 23, 26, or an allelic variant thereof.
7 . A transgenic maize plant part comprising the maize plant cell of claim 1 or 2 .
8 . The transgenic maize plant part of claim 7 , wherein said maize plant part is a seed.
9 . A transgenic maize plant comprising the maize plant cell of any one of claim 1 or 2 .
10 . A method for obtaining a bulked population of inbred seed comprising selfing the transgenic maize plant of claim 7 and harvesting seed comprising the INIR4 transgenic locus from the selfed maize plant.
11 . A method of obtaining hybrid maize seed comprising crossing the transgenic maize plant of claim 7 to a second maize plant which is genetically distinct from the first maize plant and harvesting seed comprising the INIR4 transgenic locus from the cross.
12 . A DNA molecule comprising SEQ ID NO: 8, 17, 2, 3, 8, 9, 10, 11, 16, 23, 24, 26, 27, or 28.
13 . A processed transgenic maize plant product comprising the DNA molecule of claim 12 .
14 . A biological sample containing the DNA molecule of claim 12 .
15 . A nucleic acid molecule adapted for detection of genomic DNA comprising the DNA molecule of claim 12 .
16 . The nucleic acid molecule of claim 15 , wherein said nucleic acid molecule comprises a detectable label.
17 . A method of detecting a maize plant cell comprising the INIR4 transgenic locus of claim 1 or 2 , comprising the step of detecting a DNA molecule comprising SEQ ID NO: 8, 17, 2, 3, 9, 10, 11, 16, 23, 24, 26, 27, 28, or an allelic variant thereof.
18 . A method of excising the INIR4 transgenic locus from the genome of the maize plant cell of claim 1 or 2 , comprising the steps of:
(a) contacting the INIR4 transgenic locus of the plant cell with: (i) an RNA dependent DNA endonuclease (RdDe); and (ii) a guide RNA (gRNA) capable of hybridizing to the guide RNA hybridization site of the OgRRS and the CgRRS; wherein the RdDe recognizes a OgRRS/gRNA and a CgRRS/gRNA hybridization complex; and,
(b) selecting a transgenic plant cell, transgenic plant part, or transgenic plant wherein the INIR4 transgenic locus flanked by the OgRRS and the CgRRS has been excised.
19 . The method of claim 18 , wherein the INIR4 transgenic locus of the maize plant cell comprises the CgRRS of SEQ ID NO: 8, 9, or 10.
20 . The method of claim 18 , wherein the guide RNA comprises an RNA sequence encoded by SEQ ID NO: 13.
21 . The method of claim 18 , wherein the maize plant cell comprises the INIR4 transgenic locus of SEQ ID NO: 3, 17, 27, or an allelic variant thereof.