Guangzhou Biological Institute has no trace modification of pig insulin gene, which can completely secrete human insulin.

Release date: 2016-03-30

Humanized insulin gene editing pig

On March 18th, the Journal of Molecular Cell Biology (Molecular Cell Biology) published the research results of the research group of the Chinese Academy of Sciences Guangzhou Institute of Biomedicine and Health Lai Liangxue "Genetically humanized pigs exclusively expressing human insulin are generated through custom Endonuclease-mediated seamless engineering". The study used precise gene editing technology to make a spot-free modification of the porcine insulin gene, so that the porcine insulin gene encodes human insulin, and successfully established a genetically edited pig that completely secretes human insulin.

According to the latest data released by the International Diabetes Federation in 2015, there are 415 million adults worldwide with diabetes. In 2015, 5 million people died of diabetes, exceeding the total number of deaths from malaria, tuberculosis and HIV. The cost of treatment for diabetes and related complications is enormous, estimated at more than $670 billion per year, which is higher than the full military cost of the United States.

Currently, treatment for diabetes includes insulin injections and islet transplantation. Porcine insulin has been widely used, and the use of porcine islets for xenotransplantation to treat diabetes has recently made good progress. However, compared with humans, there is an amino acid difference in insulin protein. The 30th amino acid of human insulin B chain is threonine, and porcine insulin is alanine. This difference in amino acids makes porcine insulin lower in blood glucose lowering potential in humans, and it is easy to induce antibody production in long-term use.

In the study, researchers Dr. Li Xiaoping, Dr. Yang Wei and Dr. Wang Kepin combined TALENs and CRISPR technology with single-stranded oligonucleotides to establish a unique set-point editing technique for pig genomes, using this technology to porcine insulin in somatic cells. The codon GCC of the gene encoding the 30th alanine of the B chain was modified to an ACG encoding threonine, and a homozygous cell line was obtained. At the same time, the researchers used the cell strain as a nuclear donor to successfully construct a humanized insulin cloned pig by somatic cell nuclear transfer technique, which was confirmed by high-resolution liquid chromatography tandem mass spectrometry and extracted from the genetically modified pig pancreas. Insulin is completely human insulin and does not contain porcine insulin.

The humanized insulin genetically modified pig obtained in this study will provide human insulin for the treatment of diabetes, and will also provide a more ideal donor source for clinical xenogenic islet transplantation. Technically, this achievement is also the first time to achieve a seamless genomic fixed-point modification in large animals. The establishment of this fixed-point non-marking technology will promote gene mutation and large animal disease models and genetic modification with agricultural breeding value. Animal breeding.

The research was supported by the Ministry of Science and Technology “973”, “863”, the National Natural Science Foundation, the Guangdong Provincial Science and Technology Plan Special Project and the Guangzhou Science and Technology Commission.

Source: Guangzhou Institute of Biomedicine and Health, Chinese Academy of Sciences

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