Summary of the genes tested. Wildtype gene sequences optimized for expression in different organisms<sup>1</sup> were used for embedding various messages.
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[msg] = Only codons for amino acids with ≥4 alternative codons (AGPTVLRS) were used for data storage. [msg enc] = Text message was keyed with the password “Secret” using the polyalphabetic Vigenère cipher. [msg + cut] = the codon usage table for Homo sapiens, used for encryption, was added in a 35 bp sequence after the stop codons of the ORF. [msg long] = additional codons for amino acids (CDEFHIKNQY) were used for message embedding, doubling the capacity of the container gene. Δ opt:msg = total number and percentage of altered nucleotides resulting from message deposition. CAI and GC = codon adaptation index and GC content of optimized and watermarked genes. Rel. Expression = mean relative expression ratio between watermarked and optimized genes ± standard deviation and students t-test p-value. Assay systems included expressing constructs in vivo in E. coli BL 21, S. cerevisiae strain AH109, human HEK293 cells, and in vitro with wheat germ and rabbit reticulocyte lysates; protein expression was determined by Western blots and ELISA.



