# Key Ideas from #hplusroadmap IRC Discussions

• A directed evolution approach for oligonucleotide synthesis was proposed using cells with surface-displayed enzymes, where cells improving synthesis yield would be selected while others would be flushed away, requiring at least four different cell lines (one for each nucleotide).

• Participants debated the merits of using whole cells versus isolated enzymes for directed evolution, noting that cells provide self-contained genome packages that simplify identifying genetic sequences responsible for desired functions, despite concerns about cells degrading DNA/oligonucleotides during the process.

• Alternative approaches to oligonucleotide synthesis were discussed, including Terminal deoxynucleotidyl Transferase (TdT) engineering for aqueous synthesis, which might be easier than cell-based methods but would still require tracing enzymes back to source genomes.

• The conversation explored mRNA display technology, specifically how puromycin forms a covalent link between mRNA and its translational product, with questions about the topological mechanism of how the protein-puromycin-DNA-mRNA complex escapes the ribosome.

• Antibody mimics were discussed as alternatives to traditional antibodies, particularly fibronectin type III domains and ankyrin repeat proteins, which can achieve high binding affinities without disulfide bonds, making them more versatile in different conditions.

• A protein engineering strategy was shared that involves maximizing surface area and binding variation, then selecting for minimal deletions that preserve function, resulting in smaller proteins with targeted recognition sites.

• A security vulnerability in a Bitcoin exchange was noted, specifically a flaw in two-factor authentication where verification was required for viewing but not for updating authentication settings.