The modified maize contains three gene cassettes: RNA interference (RNAi) cassette targeting Diabrotica virgifera ESCRT-III complex Snf7, a Brevibacillus laterosporus mpp75Aa1.1 cassette and a Bacillus thuringiensis vpb4Da2 cassette.

Transcription of the RNAi cassette commences from the Cauliflower mosaic virus 35S promoter and terminates at the Pisum sativum ribulose bisphosphate carboxylase small chain 2 terminator. The transcript initially contains a Zea mays heat shock protein 70 intron, which contributes to enhanced expression in vegetative tissues of the plant, and two partial coding sequences of the D. virgifera virgifera snf7 gene, which encodes the SNF7 subunit of the ESCRT-III complex. The two snf7 sequences are in an inverted orientation, separated by a 150-nucleotide intervening sequence, which allows base pairing between the inverted sequences and hairpin RNA formation post-transcription. The presence of double-stranded RNA structure then triggers an RNAi response. Due to RNAi processing, small interfering RNA molecules (roughly 21-23 nucleotides in length) will be produced and thus no translation into protein will occur from this cassette. An intergenic spacer is present between this cassette and the mpp75Aa1.1 cassette to insulate the gene cassettes (minimize effects of genetic elements between the two cassettes).

Transcription of the mpp75Aa1.1 coding sequence is under control of a Tripsacum dactyloides RCc3 promoter + leader and Coix lacryma-jobi heat shock protein 16.9 terminator. Transcription is enhanced by Setaria italica 14-3-3C intron and a Dahlia mosaic virus (DMV) enhancer. Transcription is expected in all plant tissues (no tissue-specificity).

Transcription of vpb4Da2 is under control Zea mays lipid transfer protein 1 promoter and S. italica S-adenosylmethionine synthase 1 terminator. Expression is enhanced by S. italica actin 4 intron and DMV open reading frame 6 enhancer. The promoter is not expected to be active in pollen tissues (very low or no expression).

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