Effect of knocking out hae3 gene using Cas9-CRISPR on hemoglobin of zebrafish Danio rerio
CAI Chang,WANG Cong-cong,WU Zhi-chao,LIU Yun,GUO Ya-nan,XU Qiang-hua
1. College of Marine Seienees, Shanghai Ocean University, Shanghai 201306, China; 2. Key laboratory of Sustainable Exploitation of Oceanic Fisheries Resources, Ministry of Education, Shanghai 201306, China; 3. Scientific Observing and Experimental Station of Oceanic Fishery Resources, Ministry of Agrieuhure, Shanghai201306, China; 4. National Engineering Research Center for Oceanic Fisheries, Shanghai201306, China; 5. Collaborative Innovation Center for National Distant-Water Fisheries, Shanghai 201306, China
Abstract: In order to acquire hemoglobinless mutant and investigate the function of hae3 gene during hemoglobin generation,a model of hae3 gent was constructed using Cas9-CRISPR technique in zebrafish Danio rerio model which lacks of hae3 gene.Hae3 sgRNA and Cas9 RNA mixture were injected into the animal pole of zebrafish embryo with the first cleavage stage through microinjection,and identified by sequencing,multiple sequence alignment and T7 RNase digestion methods.The hemoglobin identification was conducted by O-dianisidine staining for both the hae3 knockout group and negative control.The results showed that the expression of hemoglobin was drastically decreased in the zebrafish which lacks hae3 gene and showed deformity phenotype compared with the wild type.Thus,we validate that lacking of hae3 gene severely affects the generation of hemoglobin and leads to developmental malformation during the embryonic period.
蔡畅, 王丛丛, 吴智超, 刘云, 郭亚南, 许强华. Cas9-CRISPR敲除hae3基因对斑马鱼血红蛋白生成的影响[J]. 大连海洋大学学报, 2015, 30(6): 573-579.
CAI Chang, WANG Cong-cong, WU Zhi-chao, LIU Yun, GUO Ya-nan, XU Qiang-hua?. Effect of knocking out hae3 gene using Cas9-CRISPR on hemoglobin of zebrafish Danio rerio. Journal of Dalian Ocean University, 2015, 30(6): 573-579.
2015, Vol. 30 
