Check point kinase 2 (Chek2)為第二型損傷檢查點激酶，當細胞受到外界環境的刺激時會使Chek2活化，進而調控細胞週期，使細胞週期停滯，並且修復受損的DNA。在成功大學醫學院王家義老師實驗室的先前研究中發現除了在細胞週期的調控外，Chek2也會位在mother centriole上，而mother centriole為初級纖毛形成的要素，經由細胞培養進行的體外實驗將Chek2抑制後，發現初級纖毛的生長也會受到抑制，證明Chek2為初級纖毛生長的必要條件。因此想要探討Chek2對於體內的影響為何？在本研究中，藉由斑馬魚來觀察Chek2對於胚胎發育所造成的影響。首先確定在斑馬魚早期胚胎中有Chek2基因也有表現初級纖毛，再利用antisence Morpholino (MO)和CRISPR來抑制掉斑馬魚胚胎中Chek2的表現，觀察到斑馬魚胚胎的發育會受到影響，而且初級纖毛結構也會變得不穩定。為了加強Chek2缺失對於初級纖毛生長的證據，根據先前研究表示初級纖毛可以幫助偵測細胞外界的訊息，但是根據斑馬魚胚胎中的研究結果，Chek2的缺失並不會影響Wnt、Bmp、Nodal以及Hedgehog訊號的表現。因此經由本實驗結果得知，在水生物種及哺乳類間，Chek2調控初級纖毛生長之功能的保守性。

Checkpoint kinase 2 (Chek2) is the type II injury checkpoint kinase. When cells encounter the environmental stimulus, Chek2 will be activated and regulate the cell cycle, make the cell cycle arrest and repair damaged DNA. Therefore, Chek2 plays an important role in the regulation of cell division mechanisms. From the data of Dr. Wang’s lab in School of Medicine, National Cheng Kung University has proved that Chek2 is activated in the mother centriole, and can help the growth of primary cilia. Primary cilia is the protrusions on the cell surface, and functions that helping cells to detect the external environment stimuli and receive signals from outside to induce cell growth and differentiation. Chek2 is located on the bottom of the primary cilia and is responsible for the growth of primary cilia. However, all the data on Chek2 is from the in vitro study by using human retinal epithelial cells as a model, the function in vivo is still unknown. Therefore here explored the function of Chek2 in vivo by using zebrafish embryo as a model. First of all found that Chek2 is expressed ubiquitously during blastula and gastrula stages and mainly on the eyes and brain region at 24 hours post fertilization. Therefore, explored whether the development of eyes and the structure of primary cilia were affected by gain- or loss-of function of Chek2. Secondly, also conducted Chek2 knockout by CRISPR mutagenesis and tried to confirm the result from Chek2 knockdown. From the loss of function study it showed that the knockdown of Chek2 by morpholino resulted in the defect in eyes development and also the decrease of primary cilia growth. Finally, used Human Chek2 mRNA to rescue Chek2 knockdown zebrafish and found that the defect in eyes and the growth of primary cilia can be rescued. This rescue experiment further confirmed the Chek2 morpholino specificity. Furthermore, found Chek2 does not influence signaling pathways including Wnt, Bmp, Nodal and Hedgehog during zebrafish early embryonic development. These results suggested that the role of Chek2 might be conservative between fish and mammal that affects the primary cilia growth and the cell cycle regulation.

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