Cell-free synthesis of phage T7 improved up to tenfold with engineered E. coli lysates

Tulane University researchers used inducible CRISPR interference to manipulate 18 E. coli BL21 genes and identify effectors of phage T7 propagation, then modified the bacterial lysate used for cell-free bacteriophage synthesis accordingly. Overexpressing translation initiation factor IF-3 (infC) and the small RNAs OxyS and CyaR, or repressing the RecBCD exonuclease, raised T7 yields in vitro by up to 10-fold, which the authors say could ease phage manufacturing bottlenecks.