| Autor: |
LI Sidi, FU Bin, GUO Zhongkun, LIN Yingjie, ZHANG Zhenyu, MI Chuanliang, WANG Kezhou |
| Jazyk: |
čínština |
| Rok vydání: |
2022 |
| Předmět: |
|
| Zdroj: |
Shiyan dongwu yu bijiao yixue, Vol 42, Iss 4, Pp 294-300 (2022) |
| Druh dokumentu: |
article |
| ISSN: |
1674-5817 |
| DOI: |
10.12300/j.issn.1674-5817.2022.002 |
| Popis: |
ObjectiveTo construct a stable hereditary lipopolysaccaride binding protein (Lbp) gene knockout mice by using clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated nuclease 9 (Cas9) gene editing technology.MethodsAccording to the sequence characteristics of Lbp gene in C57BL/6J mice, the target of sgRNA was designed, the 5'-end protein coding conserved sequence of Lbp gene was deleted and the shift mutation was introduced to inactivate LBP. The genome of F0, F1, F2, F3 generation mice was extracted; PCR was used to identify and sequence Lbp knockout; RT-PCR was used to verify Lbp gene transcription, and Western blotting was used to verify LBP protein expression in F2 generation.ResultsFive Lbp+/- mice from F0 generation, three Lbp+/- mice from F1 generation, four Lbp-/- mice from F2 generation and thirty Lbp-/- mice from F3 generation were obtained. RT-PCR showed that Lbp-/- mice mRNA was 244 bp and the translation was stopped early by code-shifting mutation. Western blotting showed that LBP protein was not expressed in the liver of Lbp-/- mice.ConclusionThe Lbp gene knockout mice were successfully constructed by CRISPR/Cas9 technique, which will provide a basis for further study of the immune and physiological effects of LBP. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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