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. Author manuscript; available in PMC: 2015 Feb 21.
Published in final edited form as: Chem Soc Rev. 2013 Aug 22;43(4):1189–1200. doi: 10.1039/c3cs60208g

Figure 8.

Figure 8

a) For a FRET-CoSMoS experiment, Crawford et al. surface-tethered a pre-mRNA construct containing a FRET donor (Cy3) near the BS and a FRET acceptor (Cy5) near the 5′SS to report on pre-mRNA conformation. FRET efficiency was then correlated with the arrival or departure of labeled spliceosome subcomplexes (e.g., Atto488 labeled U1). b) Images showing the FRET efficiency (intensity of Cy5 signal) before and after the arrival of U1. Before U1 arrival (left) there is a strong fluorescent signal from the FRET acceptor (five images top left) and no signal from U1 (single image bottom left). After the arrival of U1 (single image, bottom right) there is a decrease in fluorescence intensity from the FRET acceptor. These results are consistent with the splice sites moving further apart when the pre-mRNA is occupied by U1. Images reproduced in this figure from reference 23 and with permission from the National Academy Sciences of the United States of America.