Release date: 2017-12-08
Today, scientists at the Salk Institute bring a heavyweight study – they have developed a new CRISPR-Cas9 genome editing technology that activates genes without cutting the DNA. This is expected to circumvent the main bottleneck of current gene editing technology and bring new applications. The heavy research was published today in the top academic journal Cell.
Two co-first authors of the study, Dr. Hsin-Kai Liao (left) and Dr. Fumiyuki Hatanaka (right), and the principal of the study, Professor Juan Carlos Izpisua Belmonte (Source: Salk Institute)
In recent years, the CRISPR-Cas9 system has been the focus of attention. This breakthrough technology cuts a hole in the target region of DNA to form a "double-strand break" that inserts or deletes a specific gene. However, since DNA is the key blueprint for humans to store genetic information, this practice of moving the knife on the "source code" has also caused many concerns about security.
“Although many studies have shown that CRISPR-Cas9 is an effective tool in gene therapy, there are still many people worried that it will cause unwanted mutations through double-stranded DNA breaks,†said Professor Juan Carlos Izpisua Belmonte, principal of the study. : "We can avoid such concerns."
The researchers made a subtle transformation of the CRISPR-Cas9 system. In the usual CRISPR-Cas9 system, the Cas9 protein, under the guidance of gRNA, can target specific regions of the genome, causing double-stranded DNA breaks. Recently, some scientists have also developed some new systems, in which the Cas9 protein does not have the "cut" function (inactivation). Although these Cas9 proteins bind to specific locations on the genome, they do not cause double-stranded DNA breaks. As a result, scientists have combined these proteins with other "molecular switch proteins" that affect gene expression, allowing them to act like transcriptional regulatory proteins, targeting specific genes. Although this system is creative, it has encountered bottlenecks in practical applications. These fusion proteins are too large in size to be delivered to organisms using conventional adeno-associated virus (AAV). Due to the lack of effective delivery methods, its application prospects are greatly reduced.
Research ideas of the team of Professor Izpisua Belmonte (Source: Cell)
Professor Izpisua Belmonte's team has taken a different approach. By screening a large number of combinations, they found a way to work without the need to fuse the molecular switch with the Cas9 protein (whether inactivated or not). Using different adeno-associated virus delivery vectors, researchers can deliver Cas9 proteins, molecular switches, and gRNAs into cells in batches and ensure they work.
"These different components work together in the body to affect endogenous genes," said Dr. Hsin-Kai Liao, co-first author of the study. In a sense, this new system affects the activity of genes from the "apparent" without changing the DNA sequence.
This system works in multiple organs in mice (Source: Cell)
The effect of this system was verified in mouse experiments. The researchers established a mouse model of acute kidney injury, type 1 diabetes, and a type of muscular dystrophy. In each model, they designed a CRISPR-Cas9 apparent activation system to increase the expression of specific endogenous genes. In the model of acute kidney injury, the researchers hope that the two genes expressed will be activated, producing a large number of related proteins. The kidney function of the mice has also improved; in the model of type 1 diabetes, the researchers successfully activated a gene that produces insulin-producing cells, helping mice to lower their blood sugar levels; in the last model of muscular dystrophy, Some genes that were previously confirmed to reverse the condition are activated. These results highlight the broad application prospects of this system.
“We are very excited to see these results in mice,†said Dr. Fumiyuki Hatanaka, another co-first author of the study. “We can induce gene activation and see physiological changes at the same time.â€
As planned, the team of Professor Izpisua Belmonte will further enhance the specificity of the system and apply it to more types of cells and organs to treat a wide range of human diseases.
Source: Academic Jingwei
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