Ogasawara Shinzi
Creative Research Institution Sousei, Hokkaido University , Kita 21 Nishi 10, Kita-ku, Sapporo, Hokkaido 001-0021, Japan.
PRESTO, Japan Science and Technology Agency , 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan.
ACS Chem Biol. 2017 Feb 17;12(2):351-356. doi: 10.1021/acschembio.6b00684. Epub 2017 Jan 4.
The photocontrol of protein expression enables the spatiotemporal induction of biological events in living cells or organisms. However, commonly used method such as photocontrollable transcription factor or caged nucleic acids is unsuitable for precise control of the duration of protein expression. Here, I report a photocontrollable cap (PC-cap) that can control the translation of mRNA in a reversible manner via its cis-trans photoisomerization through illumination with 370 and 430 nm light. 2-meta-Methyl-phenylazo cap (mMe-2PA-cap) in the trans form silences translation in zebrafish embryo, whereas treatment with the cis form provided a 7.1 times larger amount of translated protein compared to the trans form. Moreover, translation activated by illumination of the embryo with 370 nm light was rapidly inactivated again by subsequent illumination with 430 nm light. An application of this approach was demonstrated by photoinducing the development of double-headed zebrafish by controlling the expression period of squint protein.
蛋白质表达的光控能够在活细胞或生物体中时空诱导生物事件。然而，常用的方法如光控转录因子或笼化核酸并不适合精确控制蛋白质表达的持续时间。在此，我报道了一种光控帽（PC帽），它可以通过370和430nm光照射使其顺反异构化，以可逆的方式控制mRNA的翻译。反式的2-间甲基-苯基偶氮帽（mMe-2PA帽）使斑马鱼胚胎中的翻译沉默，而顺式处理产生的翻译蛋白量比反式多7.1倍。此外，用370nm光照射胚胎激活的翻译会在随后用430nm光照射时再次迅速失活。通过控制斜视蛋白的表达期光诱导双头斑马鱼的发育，证明了该方法的一种应用。
