Regeneration is an adaptive phenomenon with wide biological implications spread heterogeneously in almost all the organism including human beings. The ability of regeneration varies from species to species for its complexity. Epimorphic regeneration of zebrafish caudal fin tissue is the most widely studied regeneration mechanism for its discrete and rapid regenerative capability. Several genes and proteins were found to be associated with regenerative mechanisms of zebrafish caudal fin tissue. In this study we have evaluated the functional role of Annexin 2a and 2b genes during zebrafish caudal fin tissue regeneration using inventive electroporation techniques for targeting the gene involving CRISPR-Cas9 technology. The electroporation of the CRISPR was performed on the adult zebrafish caudal fin tissue post amputation. We report retarded growth during the regeneration of caudal fin tissue when Annexin 2a and 2b genes were knocked down, which was validated through gene expression & sequencing analysis and further supported by high-throughput quantitative proteomic analysis of the fin tissue. Annexin family genes such as ANXA13, ANXA1a, ANXA5b were also found to be repressed with their expression. Knocking down of ANXA2a and 2b in regenerating caudal fin tissue compromises regenerating capacity as these genes are involved in cell to cell communication and extracellular matrix growth. This study proves that ANXA2a and 2b plays a significant role in epimorphic regeneration of zebrafish caudal fin tissue.

再生是一种适应性现象，具有广泛的生物学意义，几乎在包括人类在内的所有生物中均异质分布。再生能力因物种的复杂性而异。斑马鱼尾鳍组织的表观再生是其离散和快速再生能力的最广泛研究的再生机制。发现几种基因和蛋白质与斑马鱼尾鳍组织的再生机制有关。在这项研究中，我们使用本发明的电穿孔技术靶向涉及CRISPR-Cas9技术的基因，评估了膜联蛋白2a和2b基因在斑马鱼尾鳍组织再生过程中的功能作用。截肢后，在成年斑马鱼尾鳍组织上进行CRISPR的电穿孔。我们报道了当膜联蛋白2a和2b基因被敲除时尾鳍组织再生过程中的发育迟缓，这已通过基因表达和测序分析得到验证，并得到了鳍组织高通量定量蛋白质组学的支持。膜联蛋白家族基因，例如还发现ANXA13，ANXA1a和ANXA5b的表达受到抑制。在再生尾鳍组织中敲低ANXA2a和2b会损害再生能力，因为这些基因参与细胞间的通讯和细胞外基质的生长。这项研究证明ANXA2a和2b在斑马鱼尾鳍组织的形态再生中起重要作用。