Functionalization of an Antisense Small RNA

[EN] In order to explore the possibility of adding new functions to preexisting genes, we considered a framework of riboregulation. We created a new riboregulator consisting of the reverse complement of a known riboregulator. Using computational design, we engineered a cis-repressing 5' untranslated region that can be activated by this new riboregulator. As a result, both RNAs can orthogonally trans-activate translation of their cognate, independent targets. The two riboregulators can also repress each other by antisense interaction, although not symmetrically. Our work highlights that antisense small RNAs can work as regulatory agents beyond the antisense paradigm and that, hence, they could be interfaced with other circuits used in synthetic biology. (C) 2016 The Authors. Published by Elsevier Ltd.We thank Senshi Shen and Mark Walsh for practical assistance in the laboratory, and William Rostain and John Duncan for critically reading the manuscript. This work was supported by the grants AGL2013-49919-EXP from Spanish Ministry of Economy and CSIC Intramural 201440I017 to G.R., and by the grants FP7-ICT-FET-610730 (EVOPROG), FP7-KBBE-613745 (PROMYS), EPSRC-BBSRC BB/M017982/1 (WISB centre) and startup allocation from the School of Life Sciences to A.J.Rodrigo Tarrega, G.; Prakash, S.; Cordero-Cucart, MT.; Kushwaha, M.; Jaramillo, A. (2016). Functionalization of an Antisense Small RNA. Journal of Molecular Biology. 428(5):889-892. https://doi.org/10.1016/j.jmb.2015.12.022889892428