GOTI: New detection gene editing tool off-target technology to accurately evaluate off-target rates.

"Frozen Man" (motor neurone disease), "glass doll" (bone insufficiency), "Moon Child" (alukjuonya), thalassemia... A wide variety of rare diseases have been lacking effective treatment options due to their low incidence, causing untold suffering to patients and families.
, there are more than 7,000 rare diseases in the world, 80% of which are single-gene genetic diseases.
in recent years, with the gradual maturity of gene editing technology, gene therapy has been highly anticipated.
However, the risk of gene therapy should not be underestimated, of which the "off-target effect" is the biggest risk source of gene editing technology.
recently, the Yang Hui Research Group of the Institute of Neuroscience of the Chinese Academy of Sciences and the Center for Excellence in Brain Science and Intelligent Technology teamed up with the Map Institute of Computational Biology of the Chinese Academy of Sciences, the Shenzhen Institute of Agricultural Genomics of the Chinese Academy of Agricultural Sciences and Stanford University in the United States to develop a new detection gene editing tool technology called GOTI.
the technology accurately and objectively evaluates the off-target rate of gene editing tools.
the study was published online March 1 in Science.
challenge: How to effectively detect the safety of gene editing tools CRISPR/Cas9 is a much-watched next-generation gene editing tool.
academics generally believe that clinical techniques based on CRISPR/Cas9 and its derivatives will make a significant contribution to human health.
however, the risk of "off-target" gene editing tools has also been a concern.
if it is used in clinical practice, the "off-target effect" can cause many side effects, including cancer. Yang Hui, a researcher at the Institute of Neuroscience of the Chinese Academy of Sciences in
, told China Science Daily that clinical technology has very low tolerance for potential risks and side effects, so a target detection technique that can break through previous limits will be the key to whether CRISPR/Cas9 and its derivatives can finally go clinical.
", in fact, in the past, people have introduced a variety of detection off-target programs, but these methods have limitations.
traditionally, off-target detection has relied on algorithmic predictions, unreliable uninformed, or in vitro amplification, but this introduces a lot of noise, which can lead to a significant loss of accuracy of detection.
," Yang said.
the real off-target rate of CRISPR/Cas9 and its derivatives has been controversial because of the inability to detect off-target mutations with high sensitivity, especially single nucleotide mutations.
However, any science and technology ultimately needs to serve all of humanity, especially magical technologies such as gene editing.
wanted to manipulate this "God's scalpel" effectively, and had to do a full-on medical examination.
breakthrough: GOTI technology to accurately capture the "off-target" deserter to improve the accuracy of detection of off-target effect, we must completely subvert the original off-target detection means.
to achieve this goal, the experimenters established a technology called GOTI off-target detection. "We edited a fission ball and marked it with red fluorescent proteins when the fertilized eggs of mice split into two cells,"
.
the mouse embryo developed to 14.5 days, the entire mouse embryo was digested into a single cell, using flow cell sorting technology and based on red fluorescent protein, selected gene-edited cells and cells without gene editing, and then compared the two sets of differences through genome-wide sequencing.
avoids noise problems caused by single-cell in vitro amplification.
," Zuo Diwei, a researcher at the Shenzhen Institute of Agricultural Genomics of the Chinese Academy of Agricultural Sciences, told China Science Daily.
at the same time, because the experimental group and control group came from the same fertilized egg, the theoretical genetic background is exactly the same, so directly compared to the genomes of the two groups of cells, the differences can be thought to be caused by the gene editing tool.
this allows the discovery of completely random off-target sites that were previously undetectable by off-target detection.
, the team then put the successful GOTI into gene-editing technology for off-target testing.
the experimenters first tested the most classic CRISPR/Cas9 system.
found that the well-designed CRISPR/Cas9 had no significant off-target effect.
however, the similarly highly anticipated CRISPR/Cas9 derivative be3 is very highly off target, and most of these off-target stakes occur at sites where traditional off-target predictions are considered unlikely to be off-target.
Yang Hui suggests that people should calmly analyze the security of some emerging technologies.
some of these off-target sites appear on cancer-suppressing genes, so the classic version of BE3 has a lot of hidden dangers and is not currently suitable as a clinical technique.
the future: improve gene editing treatment methods, establish industry standards Yang Hui told reporters that the team will further test BE3 in addition to causing abnormal gene mutations may exist other problems, and on this basis, try to improve the system, so as to establish a non-target, no other risk of single-base mutation technology. Li Yixue, a researcher at the MapU Institute of Computational Biology of
the Chinese Academy of Sciences, said that the latest work has established a gene editing off-target detection technology that is far superior in accuracy, breadth and accuracy, and has significantly improved the sensitivity of off-target detection in gene editing technology, which is expected to develop a new generation of gene editing tools with higher accuracy and better safety. "We hope to develop some industry standards in the future based on this new technology,"
.
any gene-editing technique entering the clinic must be tested by this system to prove its safety in order to allow the field to develop in an orderly and healthy manner.
," he said.
, a member of the Chinese Academy of Sciences and director of the Institute of Neuroscience of the Chinese Academy of Sciences, said that the technology for gene editing safety, "with it, we can more objectively and reliably assess the target rate of gene editing tools." "This allows us to re-examine the safety of gene editing technology, but it doesn't mean that it can no longer be used for gene therapy," mr.
said, referring to the major "security implications" of the technology in the single-base editing tool BE3.
it is because of new detection technologies that we know how to modify and improve BE3 to develop a new generation of more secure gene editing tools for the benefit of patients.
" related paper information: DOI: 10.1126/science.aav9973 Source: China Science Daily