Modified excisable MON87701 soybean transgenic insect resistance locus
Transgenic INIR20 soybean plants comprising modifications of the MON87701 soybean locus which provide for facile excision of the modified MON87701 transgenic locus or portions thereof, methods of making such plants, and use of such plants to facilitate breeding are disclosed.
1. A transgenic soybean plant cell comprising an INIR20 transgenic locus comprising a DNA molecule having at least 95% sequence identity across the entire length of SEQ ID NO: 3, wherein the INIR20 transgenic locus comprises an originator guide RNA recognition site (OgRRS) and a cognate guide RNA recognition site (CgRRS), and wherein said CgRRS comprises the DNA molecule set forth in SEQ ID NO: 8.
2. The transgenic soybean plant cell of claim 1 , wherein said INIR20 transgenic locus comprises the DNA molecule set forth in SEQ ID NO: 3.
3. A transgenic soybean plant part comprising the soybean plant cell of claim 1 .
4. The transgenic soybean plant part of claim 3 , wherein said soybean plant part is a seed.
5. A transgenic soybean plant comprising the soybean plant cell of claim 1 .
6. A method for obtaining a bulked population of inbred seed comprising selfing the transgenic soybean plant of claim 5 and harvesting seed comprising the INIR20 transgenic locus from the selfed soybean plant.
7. A method of obtaining hybrid soybean seed comprising crossing the transgenic soybean plant of claim 5 to a second soybean plant which is genetically distinct from the first soybean plant and harvesting seed comprising the INIR20 transgenic locus from the cross.
8. A DNA molecule comprising SEQ ID NO: 3.
9. A processed transgenic soybean plant product comprising the DNA molecule of claim 8 .
10. A biological sample containing the DNA molecule of claim 8 .
11. A method of detecting a soybean plant cell comprising the INIR20 transgenic locus of claim 1 , comprising the step of detecting a DNA molecule comprising SEQ ID NO: 8.
12. A method of excising the INIR20 transgenic locus from the genome of the soybean plant cell of claim 1 , comprising the steps of:
(a) contacting the genome of the soybean 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 of SEQ ID NO: 3; wherein the RdDe recognizes a OgRRS/gRNA and a CgRRS/gRNA hybridization complex; and,
(b) selecting a plant cell, plant part, or plant wherein the INIR20 transgenic locus flanked by the OgRRS and the CgRRS has been excised.
13. The method of claim 12 , wherein the guide RNA comprises an RNA sequence encoded by SEQ ID NO: 11.