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. 2017 Jul 12;3(7):e1700006. doi: 10.1126/sciadv.1700006

Fig. 1. Design of MGC to measure mRNA half-lives.

Fig. 1

(A) The endogenous promoter is substituted with P[tetO]4inGAL1, and silent mutations mark the RNA expressed under the control of tTA. (B) The introduction of silent mutations permits multiplexed measurements so that the expression of the endogenous gene allele is maintained to prevent the pleiotropic effects of shutting down expression. (C and D) Time course of mRNA decay in experiments using single (C) or multiplexed (D) shutdown. The two half-lives are in good agreement (1.67 and 1.66 min). Five decay curves are shown from a single multiplexed experiment pooling 10 strains. (E) The silent mutations (SM) do not alter half-lives, as evidenced by comparing the decay of mRNAs with and without silent mutations in haploid cells. The dashed gray line in the left panel indicates the expression of endogenous COG8. The gray dashed line in the right panel indicates the expected equality of the two decay rates, and the dotted lines indicate a twofold departure from this equality. Error bars represent SD (n = 3 or 4).