Method for preparing CD163 gene edited pig by using single base editor SpRY-BE4

A gene editing and single-base technology, applied in the biological field, can solve the problems of no research reports at the level of large animals, PAM limitations, etc., to improve operability and breadth, avoid random mutations and even genome damage, and improve freedom and the effect of broad
CN112094868AActive Publication Date: 2020-12-18北京首农未来生物科技有限公司
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Patent Information

Authority / Receiving Office
CN · China
Current Assignee / Owner
北京首农未来生物科技有限公司
Publication Date
2020-12-18

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Abstract

The invention discloses a method for preparing a CD163 gene edited pig by using a single base editor SpRY-BE4. By editing a CD163 gene of an isolated fibroblast genome of a pig, a 228th basic group ofan E3 exon of the CD163 diallelic gene is subjected to site-specific mutagenesis from C to T, and a TGA terminator is formed after mutagenesis, so that the expression of the E3 exon is terminated inadvance, and a CD163 diallelic gene mutant cell is obtained; and the nucleotide sequence of the E3 exon is the sequence 2. The CD163 diallelic gene knockout pig obtained by the invention has the capability of resisting PRRSV (porcine reproductive and respiratory syndrome virus), which shows that the pig resisting PRRSV can be obtained by the method for preparing the CD163 diallelic gene knockout pig.
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Description

technical field

[0001] The invention belongs to the field of biotechnology, and relates to a method for preparing CD163 gene-edited pigs by using a single base editor SpRY-BE4. Background technique

[0002] Porcine reproductive and respiratory syndrome (PRRS) is a global, highly contagious and harmful porcine viral disease with different clinical symptoms in pigs of all ages, but mainly Late abortions, stillbirths and mummies in pregnant sows and respiratory disease in piglets. Its causative agent is porcine reproductive and respiratory syndrome virus (PRRSV), and this virus mainly has two types I and II. PRRSV is a single-stranded positive-sense RNA virus, a member of the order Nidovirales, the family Arteriviridae, and the genus Arterivirus. Its genome is 15kb in length and contains 11 open reading frames. The study found that the only natural host of PRRSV is pigs, which can infect differentiated blood monocytes and porcine alveolar macrophages in pigs. Calvert et al. ...

Claims

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