Ribosome profiling vs. TRAP
I'm interested in doing ribosome profiling on a small bacterial genome. The "standard" protocols all call for isolating ribosomes with either sucrose cushions or (more recently with the Epicentre protocol) S-400 size exclusion. There seems to be an entirely different group doing ribosome isolation using affinity tags on ribosomal proteins, and they call their protocol TRAP. Both seem to get to the same place, but the affinity approaches surely seem easier and more amenable to automation. Does anyone know of why there is a reluctance to use the affinity techniques as an approach to ribosome profiling?
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CRISPR/Cas9 sparked the gene editing revolution for both research and therapeutics.1 But this system still showed severe issues that limited its applications. The most prominent were the heavy reliance on PAM sequences, delivery limitations, double-stranded breaks that prompt unintended edits and cell death, and editing inefficiency (both in targeting and in knock-in reliability).
Despite this, “CRISPR helped turn genome editing from a specialized technique into...07-31-2026, 11:01 AM