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斑马鱼基因编辑敲降服务(MO与Crispri-Dcas9)

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基因Knockdown后抑制斑马鱼血管生成

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图例. GeneX  knock down inhibits the trunk angiogenesis in zebrafish. (A-D) Representative fluorescent images of zebrafish embryos at 32h post-fertilization (hpf). (C-E) Compared with wild-type control, embryos injected with geneX-MO present a lower number of incomplete ISVs and only occasional sprouts (asterisk) of dorsal aorta. The boxed regions are shown at higher magnification in the right panels. DLAV, dorsal longitudinal anastomotic vessels; ISV, intersegmental vessels; DA, dorsal aorta; PCV, posterior cardinal vein.  

 

文献发表

Wang MS, Zhang RW, Su LY, et al. Positive selection rather than relaxation of functional constraint drives the evolution of vision duringchicken domestication. Cell Res. 2016 May;26(5):556-73.

 

 

基因Knockdown后导致中枢神经系统特异性细胞凋亡

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图例. Morpholino knock down of geneX induces potent CNS-specific apoptosis. Wild-type control embryos and embryos injected with geneX-MO were stained with acridine orange (AO) at 32hpf. Apoptotic cells are visible as black spots, and less bright homogenous black staining is unspecific background staining. (A-B) Uninjected wild-type control zebrafish exhibited few or no apoptotic cells in CNS (central nervous system). In contrast, significantly increased staining was observed throughout  the CNS in zebrafish injected with geneX-MO (C-D). The blue boxed regions are shown at higher magnification in the right panels. A-D: lateral view, anterior, left. 

 

 

 

CRISPRi

CRISPRi技术是在CRISPR/Cas9基础上,将Cas9蛋白改造成为不具有切割活性的dCas9,在sgRNA的介导下仍可以靶向基因组序列。CRISPRi技术能够有效的抑制编码基因和非编码基因的表达,并可同时对多个基因进行调控且无脱靶率。CRISPRi技术的出现为研究基因的功能提供了高效的新方法。