Targeted insertion via transposition

An engineered nucleic acid modification system for generating a genetically modified soybean cell, the system comprising:

• a. a donor polynucleotide comprising a first and second mPing miniature inverted-repeat transposable element (MITE) transposition sequences;
• b. one or more nucleic acid constructs for expressing
• i. a Pong ORF1 protein of a Pong transposase, the nucleic acid construct comprising a promoter operably linked to a nucleic acid sequence encoding the Pong ORF1 protein;
• ii. a nucleic acid construct for expressing a targeting nuclease of a CRISPR/Cas system comprising a Cas nuclease and a guide RNA (gRNA), wherein the Cas nuclease is fused to the C-terminus of a Pong ORF2 protein of the Pong transposase by more than one copy of a flexible linker, the nucleic acid construct comprising a promoter operably linked to a nucleic acid sequence encoding the Cas nuclease fused to the C-terminus of the Pong ORF2 protein by more than one copy of a flexible linker;
• and
• iii. a nucleic acid expression construct for expressing the gRNA of the CRISPR/Cas system;
• wherein the CRISPR/Cas system is programmed to target the transposase and the donor polynucleotide to a target nucleic acid locus in the cell, to introduce a cut in the target nucleic acid locus, or both, thereby accomplishing insertion of the donor polynucleotide at the target nucleic acid locus to generate a genetically modified cell comprising the donor polynucleotide inserted at the target nucleic acid locus.