Small molecule composition for partial reprogramming of pig somatic cells, partial reprogramming nuclear donor cell and application of partial reprogramming nuclear donor cell

By screening small molecule combinations of CHIR99021, Repsox-616452, TTNPB, and SAG suitable for porcine "3+X" gene-edited cells, partial reprogramming was performed, solving the problem of deterioration in nuclear donor cell quality and achieving efficient and safe nuclear donor cell preparation, significantly reducing early abortion rate and improving production efficiency.

CN122146618APending Publication Date: 2026-06-05ZHUHAI HENGQIN ONA REGENERATIVE MEDICINE CO LTD
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Patent Information

Application Number
CN202610436499.3
Authority / Receiving Office
CN · China
Patent Type
Applications(China)
Current Assignee / Owner
Filing Date
2026-04-02
Publication Date
2026-06-05

AI Technical Summary

Technical Problem

Existing small molecule reprogramming technology cannot effectively improve the quality of "3+X" gene-edited pig somatic cells, leading to nuclear donor cell aging and incomplete nuclear reprogramming, which in turn causes problems such as embryo implantation failure and high early miscarriage rates. Furthermore, it suffers from species-specific barriers, lack of targeted protocols, and uncontrollable safety.

Method used

Using a small molecule composition of CHIR99021, Repsox-616452, TTNPB and SAG, at specific concentrations and ratios, porcine "3+X" gene-edited cells were partially reprogrammed to reverse their aging-related phenotypic and functional states, thus producing high-quality nuclear donor cells.

Benefits of technology

It significantly reduced the early abortion rate to below 10%, improved the clonal expansion potential and production efficiency of cells, solved the bottleneck in the preparation of a moderate number of genetically modified donor pigs, and achieved a high pregnancy maintenance rate and healthy farrowing rate.

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Abstract

The invention provides a small molecule composition for partial reprogramming of pig somatic cells, a partial reprogramming nuclear donor cell and application of the partial reprogramming nuclear donor cell and belongs to the field of animal biotechnology and genetic engineering, and the small molecule composition comprises CHIR99021, Repsox-616452, TTNPB and SAG. The small molecule composition provided by the invention can safely and obviously reverse the appearance and functional aging state of pig aging somatic cells edited by '3 + X', recover the compartment integrity of the nucleoplasm region, and improve the cell cloning and amplification capability by 200% or more. The obtained partially reprogrammed nuclear donor cells are used as nuclear donors for SCNT, the early abortion rate can be reduced from more than 70% to less than 10%, and the industry limit level is reached. The invention provides an efficient key technology for large-scale and economic production of the'gold standard 'donor pigs.
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