Dissecting the Genetic Basis of a Complex cis-Regulatory Adaptation
Fig 4
Effects of candidate gene deletion and over-expression.
A. The five candidate genes were deleted from Sp, and the effect on resistance to 300 ppm citrinin was measured. Four deletions increased sensitivity (t-test p < 0.005 for each), and were studied further. Error bars show 1 S.E. B. The effects of three different dCas9 fusion proteins targeted to four candidate genes were measured by qPCR. C. Illustration of our strategy to over-express four genes via dCas9 fusion protein. D. We used direct competition for 40 generations to measure the relative fitness of the four-gene over-expression strain (with a Gal4-dCas9-VP64 dual fusion) vs. a control strain containing the same plasmid, but lacking any gRNAs. Conditions were YPD, and YPD + 300 ppm citrinin. Error bars show 1 S.E.