Characterization of biomodal light-switchable rhodopsins and tailoring for optogentic application

黑视素是哺乳动物中负责非视觉光感受和控制过程如瞳孔收缩和昼夜节律的光感受器。迄今已知的哺乳动物（小鼠）的黑视素和一些斑马鱼黑视素是光致变色的，并且在暗态和信号传导态之间是双峰可切换的，这是光遗传学应用最感兴趣的特性。在这个项目中，我们将在HEK细胞，COPS细胞和毕赤酵母中表达几种黑视素，并在UV/维斯和振动光谱中进行研究，时间范围从fs到秒（与阿姆斯特丹的Tilo Mathes合作）。对于光遗传学应用，我们将产生光致变色黑视素和GPCR型激素受体的杂交体，目的是用光激活激素受体信号传导。我们将与针对促肾上腺皮质激素释放激素（CRH）的Soojin Ryu合作验证这一概念。对于这种方法，我们将纯化和纯化七鳃鳗的Parapinopsin和斑马鱼的opn 4。我们交换的激素受体的互补环的细胞内环和测试的功能，在鱼苗与Soojin Ryu。

Melanopsins are Photoreceptors that are in mamals responsible for non-viual photoreception and control processes like pupil constriction and circadian rhythm. The so far known Melanopsins of mamals (mouse) and some of the zebrafish melanopsins are photochromic and bimodal switchable between the dark state and the signaling state, a property that is of highest interest for optogenetic application. In this project we will express several melanopsins in HEK-cells, COPS cells and in Pichia pastoris and carry out studies in the UV/Vis and vibrational spectroscopy on time scales ranging from fs to seconds (with Tilo Mathes in Amsterdam). For optogenetic applications we will produce hybrids of photochromic melanopsins and GPCR-type hormone receptors with the intention to activate hormone receptor signalling with light. We will validate this concept in collaboration with Soojin Ryu targeting corticotrophin releasing hormone (CRH). For this approach we will purify and characterise Parapinopsin of lamprey and opn4 of zebrafish. We exchange the intracellular loops against the complementary loops of the hormone receptor and test the function in fish larvae with Soojin Ryu.