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This page was last modified on 27 Jan 2025 27 Jan 2025 .
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The amount of proteins present in cells of living organisms depends on various factors, including the rate of transcription of coding sequences, the efficiency with which pre-mRNA transcripts are processed and exported from the nucleus to the cytoplasm, the translatability of a transcript, and the stability of the mRNA. Research has demonstrated that addition of introns to coding sequences can significantly increase eukaryotic in vivo expression levels. We present here a protocol for inserting introns into coding sequences of interest to optimize expression. This protocol consists of the following steps: (1) codon-optimizing the coding sequence of interest, (2) defining positions for intron insertion, (3) selecting introns to be introduced, (4) in silico insertion of introns into coding sequences followed by sequence optimization, (5) finalizing the sequence design by addition of flanking restriction sites, and (6) ordering the sequences from a gene synthesis provider.
This page was last modified on 27 Jan 2025 27 Jan 2025 .

