The Dr. John and Anne Chong Lab for Functional Genomics, Charles Perkins Centre and School of Life & Environmental Sciences, The University of Sydney, Sydney, NSW, 2006, Australia.
Genome Editing Initiative, The University of Sydney, Sydney, NSW, 2006, Australia.
Nat Commun. 2019 Apr 30;10(1):1655. doi: 10.1038/s41467-019-09681-1.
The box jellyfish Chironex fleckeri is extremely venomous, and envenoming causes tissue necrosis, extreme pain and death within minutes after severe exposure. Despite rapid and potent venom action, basic mechanistic insight is lacking. Here we perform molecular dissection of a jellyfish venom-induced cell death pathway by screening for host components required for venom exposure-induced cell death using genome-scale lenti-CRISPR mutagenesis. We identify the peripheral membrane protein ATP2B1, a calcium transporting ATPase, as one host factor required for venom cytotoxicity. Targeting ATP2B1 prevents venom action and confers long lasting protection. Informatics analysis of host genes required for venom cytotoxicity reveal pathways not previously implicated in cell death. We also discover a venom antidote that functions up to 15 minutes after exposure and suppresses tissue necrosis and pain in mice. These results highlight the power of whole genome CRISPR screening to investigate venom mechanisms of action and to rapidly identify new medicines.
箱型水母 Chironex fleckeri 毒性极强，在严重暴露后的几分钟内，毒液会导致组织坏死、极度疼痛和死亡。尽管毒液作用迅速而强烈，但基本的机制仍不清楚。本研究通过筛选大规模 lenti-CRISPR 诱变的宿主成分，对水母毒液诱导的细胞死亡途径进行分子剖析，发现外周膜蛋白 ATP2B1（一种钙转运 ATP 酶）是毒液细胞毒性所必需的宿主因子之一。靶向 ATP2B1 可阻止毒液作用并提供持久的保护。对毒液细胞毒性所必需的宿主基因进行计算分析，揭示了以前未涉及细胞死亡的途径。我们还发现了一种毒液解毒剂，在暴露后 15 分钟内仍有效，并能抑制小鼠的组织坏死和疼痛。这些结果突出了全基因组 CRISPR 筛选在研究毒液作用机制和快速发现新药方面的强大功能。
