论文
论文标题: Creation of intermuscular bone-free mutants in amphitriploid gibel carp by editing two duplicated runx2b homeologs
作者: Gan, Rui-Hai; Li, Zhi; Wang, Zhong-Wei; Li, Xi-Yin; Wang, Yang; Zhang, Xiao-Juan; Tong, Jin-Feng; Wu, Yue; Xia, Ling-Yun; Gao, Ze-Xia; Zhou, Li; Gui, Jian-Fang
出版刊物: AQUACULTURE
出版日期: MAR 30
出版年份: 2023
卷/期:
DOI: 10.1016/j.aquaculture.2023.739300
论文摘要: Intermuscular bones (IBs) are ubiquitous in most of freshwater farmed fishes, especially in cyprinid species, and the creation of IB-free mutants has become a hot issue in fish genetic breeding because of the potential beneficial for consumers and fish processing industry. Recently, significant progress in the mechanism underlying IB development has been achieved in zebrafish and a decisive player, runt-related transcription factor 2b (runx2b), has been identified. However, its role has not yet been validated in other farmed fishes. Amphitriploid gibel carp (Carassius gibelio) is an important aquaculture species in China, but the injuries threats with approximately 80 IBs have strongly restricted its industry development. In this study, we first identified two homeologs of runx2b (Cgrunx2b-A and Cgrunx2b-B) in gibel carp with about 91.8% identity, each of them possessing three alleles with 99.6% identities. Then, we traced the IB ossification process. Alizarin red S staining showed that gibel carp IB began to ossify at 14 approximately days post hatching (dph) and developed from the posterior to the anterior regions. Cgrunx2b-A and Cgrunx2b-B displayed similar expression pattern but differential expression level during IB development. Moreover, we applied an optimized protocol of CRISPR/Cas9 editing in the polyploid Carassius complex to disrupt the functions of CgRunx2b-A and CgRunx2b-B singly or simultaneously. IB development was unaffected by single deficiency of CgRunx2b-A or CgRunx2b-B, but simultaneous disruption of all alleles of both CgRunx2b-A and CgRunx2b-B leads to complete loss of IB. The results indicate that CgRunx2b-A and CgRunx2b-B cooperatively regulate IB development. Importantly, we created several gynogenetic gibel carp mutants, including IB-free Cgrunx2b-A / / ;Cgrunx2b-B / / mutants and IB-less Cgrunx2b-A / / ;Cgrunx2b-B / /+/- mutants. The current study not only confirms the cooperatively decisive role of two homeologs of runx2b in IB formation, but also provides a feasible gene editing-based approach to create IB-free mutants in gibel carp.
== 实验室与学会 ==
  • == 实验室与学会 ==
  • 淡水生态与生物技术国家重点实验室
  • 中国科学院藻类生物学重点实验室
  • 农业部淡水养殖病害防治重点实验室
  • 武汉白暨豚保护基金会
  • 湖北省海洋湖沼学会
  • 中国动物学会原生动物学分会
  • 中国动物学会斑马鱼分会
  • 湖北省暨武汉动物学会
  • 中国水产学会鱼病学专业委员会
  • 中国鱼类学会
== 平台建设 ==
  • == 平台建设 ==
  • “一带一路”海域赤潮数据库
  • 国家水生生物种质资源库
  • 国家斑马鱼资源中心
  • 中国科学院淡水藻种库
  • 中国科学院武汉生命科学大型仪器区域中心
  • 湿地生态系统观测研究野外站联盟
  • 中国科学院水生生物研究所分析测试中心
  • 中国科学院超级计算武汉分中心
  • 水生生物博物馆
== 相关网站推荐 ==
  • == 相关网站推荐 ==
  • 中国科学院
  • 农业农村部
  • 科学技术部
  • 生态环境部
  • 国家自然科学基金委员会
  • 中国科学院武汉分院
  • 湖北省科学技术厅
  • 湖北省生态环境厅
  • 湖北省农业农村厅