FnCas12a/crRNA-Mediated Genome Editing in Eimeria tenella

• DOI: 10.3389/fgene.2021.738746
• Publication Date: 2021-09-22T05:34:16Z
• Authors: Peipei Cheng; Zhihao Zhang; Fayu Yang; Shuo Cai; Lina Wang; Chunmei Wang; Mi Wang; Yingchun Liu; Chenzhong Fei; Lifang Zhang; Feiqun Xue; Feng Gu
• Keywords: 0303 health sciences; 03 medical and health sciences; Genetics; genome editing; EtActin; QH426-470; FnCas12a; knock-in; Eimeria tenella; EtHistone H4; 3. Good health

Eimeria species are intracellular parasites residing inside the intestinal epithelial cell, which cause poultry coccidiosis and result in significant financial losses industry. Genome editing of is immense importance for development vaccines drugs. CRISPR/Cas9 has been utilized manipulating genome tenella ( E. ). Ectopic expression Cas9, i.e., via plasmids, would introduce transgene, substantially limits its application, especially vaccine development. In this study, we initially optimized condition transfection protocol. We demonstrated that with condition, FnCas12a (also known as “FnCpf1”) protein crRNA targeting EtHistone H4 triggered DNA double-strand breaks vivo . then used strategy to knock-in a coding cassette an enhanced yellow fluorescent EYFP ) dihydrofolate reductase–thymidylate synthase gene DHFR selection marker tag endogenous EtActin The engineered parasite possesses entire life cycle. Our results could trigger , augments applicability dissection function anticoccidial drugs species.