Sci Adv: East China Shishi Adeshi Ye Haifeng Team: Developing a Far Red Light-Activated Gene Editing System - FAST

Introduction !----: The rapid development of gene editing technology has provided more and more tools for gene functional research, the CRISPR-Cas system is the most widely used gene editing toolrecently, the team of Ye Haifeng of East China Normal University has designed a far-red-light-activated Split-Cas9 gene editing system that enables remote gene editing of organs and tumorscrispR-Cas9 system as a revolutionary gene-editing technology, its convenience offers unprecedented opportunities for scientific research and disease treatment, including high-throughput screening, functional genomics research, and the treatment of viral infections, genetic diseases, and cancershowever, it is well known that the CRISPR-Cas9 system has some disadvantages, such as off-target effectsscientists are also studying how to overcome these shortcomingsrecently, Chinese scientists have developed a split-Cas9 gene editing system activated by far red light (FRL), which can freely induce gene editing in cells deep in animal tissuethe study, led by Researcher Ye Haifeng of the School of Life Sciences of East China Normal University and published in Science Advances, entitled "Engineering a far-red light-activated-split-cas9 system for remote-controlled-genome siting of organs and tumors" has previously been developed by scientists who have developed a variety of chemically induced CRISPR-Cas9 systems, the notable drawback of which is the potential toxicity of these chemical inducers, scientists have used light as a reversible, noninvasive way to induce to develop Cas9-based gene editing techniquessuch as Blu-ray-based paCas9 systems, uv-mediated complementary oligonucleotide element cutting, and Blue Light-based anti-CRISPR systemsHowever, neither ultraviolet nor blue light can penetrate the body because of the absorption and scattering of these light by biological tissues, as a result, in this study, Ye Haifeng's team used far red light (730 nm)in 2017, Ye Haifeng's research team developed a far-red light regulatory genetically modified expression control systemachieves a single beam of distant red light to regulate gene expressionuses multidisciplinary technology to cross-establish an electronic drug platform for the treatment of diabetes through the super-remote regulation of artificial islet cells through the smartphone APPMobile teletherapeutic diabetes design schematics In 2018, they combined the far red light regulatory genetically modified expression control system with crispR-dCas9 gene editing technology to develop the CRISPR-dCas9 endogenous gene transcription activation device (FACE) of far red light regulationthe CRISPR-dCas9 endogenous gene transcription activation device (FACE) for far red light control this time they developed a far-red light-activated Split-Cas9 (FAST) system that can uninvasively induce gene editing activity in cells deep inside animal tissuetheFAST system is designed by two split Cas9 fusion proteins with high affinity binding domains: half of Cas9 is composed expression, while the other half is LED-induced by the team's previously established bacterial photosensitive pigment BphS optical control systeminitially, researchers placed FAST system components in human embryonic kidney (HEK)-293 cells and used FRL lighting based on light-emitting diodes (LEDs) to demonstrate successful activation of targeted genome editingAfter, FRL-induced editing was achieved in different human cell lines, and the researchers demonstrated through implant experiments that FAST was able to powerfully activate gene editing in cells located in the animal's subcutaneous tissueFRL-induced FAST-mediated mice implanted in subcutaneous cells, and then, in an experiment with the genetically modified tdDTOMATO report ingon, they established the rapid mediated editing of FRL-induced mouse somatic cells (liver cells) and the study of cell cycle inactivation gene editing of cancer cells in mice with xenotransplanttumor tumors, to demonstrate that FAST can fight diseaseFAST-mediated tdTomato reportgene-editing fast system in hetero-transplanted mice in tumor gene editing this study expanded optogenetic tools for mammalian cell gene editing, including energy induction of FRL in vivo compatibility and deep tissue penetration, and demonstrated the application of this technique in relation to basic biology and biomedical research