US 12,404,515 B2
Simultaneous gene editing and haploid induction
Timothy Kelliher, Durham, NC (US); and Qiudeng Que, Durham, NC (US)
Assigned to Syngenta Crop Protection AG, Basel (CH)
Appl. No. 17/327,016
Filed by SYNGENTA PARTICIPATIONS AG, Basel (CH); and SYNGENTA CROP PROTECTION, LLC, Greensboro, NC (US)
PCT Filed Dec. 17, 2019, PCT No. PCT/US2019/066719
§ 371(c)(1), (2) Date May 21, 2021,
PCT Pub. No. WO2020/131788, PCT Pub. Date Jun. 25, 2020.
Claims priority of provisional application 62/783,446, filed on Dec. 21, 2018.
Prior Publication US 2023/0060937 A1, Mar. 2, 2023
Int. Cl. C12N 15/82 (2006.01); A01H 1/02 (2006.01); A01H 1/04 (2006.01); C12Q 1/6895 (2018.01)
CPC C12N 15/8213 (2013.01) [A01H 1/021 (2021.01); A01H 1/045 (2021.01); C12Q 1/6895 (2013.01); C12Q 2600/13 (2013.01); C12Q 2600/158 (2013.01); C12Q 2600/172 (2013.01)] 9 Claims
OG exemplary drawing
 
1. A method of testing an edited haploid maize plant progeny for the absence of a first maize plant's genomic material, wherein the first maize plant is an NP2222 haploid inducer line, the method comprising:
isolating a nucleic acid from the edited haploid maize progeny, and detecting in the nucleic acid the presence of a plurality of codominant markers, wherein the codominant markers have a distinct haplotype for a second maize plant, wherein the second maize plant is a B14 line, and wherein the second maize plant comprises the source of genomic DNA in the edited haploid maize plant progeny,
wherein the plurality of codominant markers includes SEQ ID NOs 474-548, and
wherein the detecting includes the use of at least one set of primers and at least one probe selected from SEQ ID NOs: 174-473,
wherein SEQ ID NOs: 174, 175, and 177 are used to detect SEQ ID NO: 474, SEQ ID NOs: 178, 179, and 181 are used to detect SEQ ID NO: 475, SEQ ID NOs: 182, 183, and 184 are used to detect SEQ ID NO: 476, SEQ ID NOs: 186, 187, and 189 are used to detect SEQ ID NO: 477, SEQ ID NOs: 190, 191, and 193 are used to detect SEQ ID NO: 478, SEQ ID NOs: 194, 195, and 197 are used to detect SEQ ID NO: 479, SEQ ID NOs: 198, 199, and 200 are used to detect SEQ ID NO: 480, SEQ ID NOs: 202, 203, and 205 are used to detect SEQ ID NO: 481, SEQ ID NOs: 206, 207, and 208 are used to detect SEQ ID NO: 482, SEQ ID NOs: 210, 211, and 213 are used to detect SEQ ID NO: 483, SEQ ID NOs: 214, 215, and 217 are used to detect SEQ ID NO: 484, SEQ ID NOs: 218, 219, and 220 are used to detect SEQ ID NO: 485, SEQ ID NOs: 222, 223, and 224 are used to detect SEQ ID NO: 486, SEQ ID NOs: 226, 227, and 228 are used to detect SEQ ID NO: 487, SEQ ID NOs: 230, 231, and 232 are used to detect SEQ ID NO: 488, SEQ ID NOs: 234, 235, and 236 are used to detect SEQ ID NO: 489, SEQ ID NOs: 238, 239, and 241 are used to detect SEQ ID NO: 490, SEQ ID NOs: 242, 243, and 245 are used to detect SEQ ID NO: 491, SEQ ID NOs: 246, 247, and 249 are used to detect SEQ ID NO: 492, SEQ ID NOs: 250, 251, and 253 are used to detect SEQ ID NO: 493, SEQ ID NOs: 254, 255, and 257 are used to detect SEQ ID NO: 494, SEQ ID NOs: 258, 259, and 261 are used to detect SEQ ID NO: 495, SEQ ID NOs: 262, 263, and 264 are used to detect SEQ ID NO: 496, SEQ ID NOs: 266, 267, and 269 are used to detect SEQ ID NO: 497, SEQ ID NOs: 270, 271, and 273 are used to detect SEQ ID NO: 498, SEQ ID NOs: 274, 275, and 277 are used to detect SEQ ID NO: 499, SEQ ID NOs: 278, 279, and 281 are used to detect SEQ ID NO: 500, SEQ ID NOs: 282, 283, and 284 are used to detect SEQ ID NO: 501, SEQ ID NOs: 286, 287, and 288 are used to detect SEQ ID NO: 502, SEQ ID NOs: 290, 291, and 293 are used to detect SEQ ID NO: 503, SEQ ID NOs: 294, 295, and 296 are used to detect SEQ ID NO: 504, SEQ ID NOs: 298, 299, and 300 are used to detect SEQ ID NO: 505, SEQ ID NOs: 302, 303, and 304 are used to detect SEQ ID NO: 506, SEQ ID NOs: 306, 307, and 309 are used to detect SEQ ID NO: 507, SEQ ID NOs: 310, 311, and 313 are used to detect SEQ ID NO: 508, SEQ ID NOs: 314, 315, and 316 are used to detect SEQ ID NO: 509, SEQ ID NOs: 318, 319, and 320 are used to detect SEQ ID NO: 510, SEQ ID NOs: 322, 323, and 325 are used to detect SEQ ID NO: 511, SEQ ID NOs: 326, 327, and 329 are used to detect SEQ ID NO: 512, SEQ ID NOs: 330, 331, and 333 are used to detect SEQ ID NO: 513, SEQ ID NOs: 334, 335, and 336 are used to detect SEQ ID NO: 514, SEQ ID NOs: 338, 339, and 341 are used to detect SEQ ID NO: 515, SEQ ID NOs: 342, 343, and 344 are used to detect SEQ ID NO: 516, SEQ ID NOs: 346, 347, and 348 are used to detect SEQ ID NO: 517, SEQ ID NOs: 350, 351, and 352 are used to detect SEQ ID NO: 518, SEQ ID NOs: 354, 355, and 356 are used to detect SEQ ID NO: 519, SEQ ID NOs: 358, 359, and 361 are used to detect SEQ ID NO: 520, SEQ ID NOs: 362, 363, and 365 are used to detect SEQ ID NO: 521, SEQ ID NOs: 366, 367, and 368 are used to detect SEQ ID NO: 522, SEQ ID NOs: 370, 371, and 372 are used to detect SEQ ID NO: 523, SEQ ID NOs: 374, 375, and 376 are used to detect SEQ ID NO: 524, SEQ ID NOs: 378, 379, and 38 are used to detect SEQ ID NO: 525, SEQ ID NOs: 382, 383, and 384 are used to detect SEQ ID NO: 526, SEQ ID NOs: 386, 387, and 388 are used to detect SEQ ID NO: 527, SEQ ID NOs: 390, 391, and 393 are used to detect SEQ ID NO: 528, SEQ ID NOs: 394, 395, and 397 are used to detect SEQ ID NO: 529, SEQ ID NOs: 398, 399, and 400 are used to detect SEQ ID NO: 530, SEQ ID NOS: 402, 403, and 404 are used to detect SEQ ID NO: 531, SEQ ID NOs: 406, 407, and 409 are used to detect SEQ ID NO: 532, SEQ ID NOs: 410, 411, and 412 are used to detect SEQ ID NO: 533, SEQ ID NOs: 414, 415, and 416 are used to detect SEQ ID NO: 534, SEQ ID NOS: 418, 419, and 420 are used to detect SEQ ID NO: 535, SEQ ID NOs: 422, 423, and 425 are used to detect SEQ ID NO: 536, SEQ ID NOs: 426, 427, and 429 are used to detect SEQ ID NO: 537, SEQ ID NOs: 430, 431, and 433 are used to detect SEQ ID NO: 538, SEQ ID NOS: 434, 435, and 437 are used to detect SEQ ID NO: 539, SEQ ID NOs: 438, 439, and 440 are used to detect SEQ ID NO: 540, SEQ ID NOs: 442, 443, and 445 are used to detect SEQ ID NO: 541, SEQ ID NOs: 446, 447, and 449 are used to detect SEQ ID NO: 542, SEQ ID NOS: 450, 451, and 453 are used to detect SEQ ID NO: 543, SEQ ID NOs: 454, 455, and 456 are used to detect SEQ ID NO: 544, SEQ ID NOs: 458, 459, and 461 are used to detect SEQ ID NO: 545, SEQ ID NOs: 462, 463, and 465 are used to detect SEQ ID NO: 546, SEQ ID NOS: 466, 467, and 468 are used to detect SEQ ID NO: 547, and SEQ ID NOs: 470, 471, and 473 are used to detect SEQ ID NO: 548.