Animal, Dairy, and Veterinary Sciences Department, College of Agriculture and Applied Sciences, Utah State University, Logan, UT, 84322, USA.
Moores Cancer Center, University of California, San Diego, La Jolla, CA, 92093, USA.
Sci Rep. 2024 Feb 29;14(1):4987. doi: 10.1038/s41598-024-55464-0.
Allergens from domestic cats (Felis catus) cause allergy-related health problems worldwide. Fel d 1 is a major allergen that causes severe allergic reactions in humans, including rhinitis, conjunctivitis, and life-threatening asthma. Therefore, patients with cat allergies anticipate hypoallergenic cats. We successfully generated Fel d 1 chain 2 (CH2) genome-edited cats using the CRISPR-Cas9 system in this study. T7 endonuclease 1 assay and Sanger sequencing were used to confirm the mutation in CH2 genome-edited cats. Fel d 1 level in CH2 genome-edited cats were assessed by enzyme-linked immunosorbent assay (ELISA). Remarkably, ELISA showed that the level of Fel d 1 in the CH2 homozygous genome-edited cat (Name: Alsik) was extremely low compared with that in wild type domestic cats and could be hypoallergenic cats. Additionally, we successfully cloned the CH2 homozygous genome-edited cat using cytoplasm injection clone technology. The cloned CH2 homozygous genome-edited cat was verified using microsatellite analysis. Creating hypoallergenic cats using the CRISPR-Cas9 system is a significant step forward because these cats can safely approach allergic patients.
家猫（Felis catus）过敏原在全球范围内引起与过敏相关的健康问题。Fel d 1 是一种主要过敏原，可导致人类发生严重过敏反应，包括鼻炎、结膜炎和危及生命的哮喘。因此，对猫过敏的患者期待低过敏猫。在这项研究中，我们使用 CRISPR-Cas9 系统成功生成了 Fel d 1 链 2（CH2）基因编辑猫。使用 T7 内切酶 1 分析和 Sanger 测序来确认 CH2 基因编辑猫中的突变。通过酶联免疫吸附试验（ELISA）评估 CH2 基因编辑猫中的 Fel d 1 水平。值得注意的是，ELISA 表明，CH2 纯合基因编辑猫（名称：Alsik）中的 Fel d 1 水平与野生型家猫相比极低，可能是低过敏猫。此外，我们使用细胞质注射克隆技术成功克隆了 CH2 纯合基因编辑猫。使用微卫星分析验证了克隆的 CH2 纯合基因编辑猫。使用 CRISPR-Cas9 系统创建低过敏猫是向前迈出的重要一步，因为这些猫可以安全地接近过敏患者。
