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Professor Han Xin's team from Nanjing University of Chinese Medicine Small: Development of spatio-temporal specific regulation of gene editing technology and its application in sensitive and enhanced tumor photothermal therapy
The new generation of CRISPR gene editing technology has significant advantages such as high efficiency, easy operation, and low cost. It can carry out accurate and stable genetic modification of genome sequences and bring revolutionary changes to life sciences and clinical treatment. However, due to the large immunogenicity, genotoxicity, and uncontrollable off-target effects, gene editing therapy is often accompanied by serious and unpredictable side effects; in addition, how to precisely target specific cell groups and induce them is effective for treatment The high level of gene editing and the establishment of a replicable and scalable production process are the key factors and technical bottlenecks that restrict the ultimate clinical application of gene editing therapy.
In response to the above key issues, Professor Han Xin's team at Nanjing University of Traditional Chinese Medicine has developed a spatio-temporal specific regulation of gene editing technology to achieve precise regulation of near-infrared light gene editing and drug release, and successfully applied this technology to malignant tumors including gene therapy The synergistic combination therapy of, light, heat and low temperature sensitive therapy and chemotherapy provides new exogenous controllable strategies and methods to improve gene editing efficiency while reducing off-target effects, and provides new ideas for the application of gene editing technology in disease treatment .
This study uses the excellent light-to-heat conversion material CuS nanoparticles as a tool to deliver the gene editing element Cas9 ribonucleoprotein complex (Cas9 RNP), and realizes the effective load of Cas9 RNP through complementary pairing of ssDNA and sgRNA on the surface of CuS nanoparticles. Intracellular delivery. The chemotherapeutic drug Doxorubicin (DOX) interacts with double-stranded DNA molecules. The anthracycline chromophore of DOX can be inserted into the GC-CG position of DNA. Therefore, the gene editing delivery system can also realize the loading of chemotherapeutic drugs at the same time. Strictly controlling the gene expression process and realizing precise gene editing with temporal and spatial specificity are essential to improve the specificity and safety of gene therapy. The nano delivery system reported in this work has the following highlights:
①CuS nanoparticles act as a "photothermal converter", which can stably convert near-infrared light (808 nm) into a local thermal effect to provide photothermal stimulation; ②By designing the length of complementary base pairs in the ssDNA/sgRNA hybrid chain to precisely control it Melting temperature to achieve controllable release of sgRNA and DOX; ③Through targeted knockout of heat shock protein Hsp90α to reduce the thermal tolerance of tumor cells, enhance the effect of photothermal therapy, and achieve two-way synergistic inhibition of photothermal and molecular targeting Effect. This kind of exogenously controllable, conditionally responsive gene editing and drug release provides new breakthroughs and technical reference for the precise and coordinated combination therapy of tumors.
This work was selected as a Front Cover by Small and published. Nanjing University of Traditional Chinese Medicine is the first communication unit. PhD student Chen Chao is the first author of this paper, Professor Han Xin is the corresponding author, and Professor Zha Zhengbao of Hefei University of Technology He and Professor Song Yujun of Nanjing University are the co-corresponding authors. This research has been supported by the National Natural Science Foundation of China (31901010, 21874066), Jiangsu University Advantage Discipline Construction Project (Integrated Traditional Chinese and Western Medicine), Jiangsu Specially-appointed Professor Natural Science Funded Talent Project, Supported by Jiangsu Province's "Entrepreneurship and Innovation Program" team projects.
The above work is another important achievement based on a series of research results obtained by Professor Han Xin based on the combination of microfluidic chip technology and gene editing technology. Professor Han Xin previously developed a novel microfluidic chip cell transfection technology, which successfully overcomes the bottleneck of molecular delivery and gene editing in difficult-to-transfect cells such as T cells and hematopoietic stem cells, and provides new opportunities for the clinical application of tumor immunotherapy and other cell therapies. (Han X, et al. Sci. Adv., 2015, 1, e1500454); developed a high-throughput microfluidic cell sorting chip, combined with CRISPR technology to achieve rapid and effective tumor molecular targets throughout the genome Screening and identification (Han X, et al. Angew. Chem. Int. Ed., 2016, 55, 8561-8565). The micro-nano manufacturing and translational medicine cross-team laboratory (https://www.hanlabnj.com/) where Professor Han Xin’s team is located is committed to using a new generation of gene editing technology, novel microfluidic chip technology and multi-functional intelligent biomaterials to solve The key scientific and technical issues in the biomedical transformation process. The team members have published more than 10 high-level research papers such as Science Advances, Angewandte Chemie, Advanced Materials, Advanced Functional Materials, Small, Nano Letters, Biomaterials, etc. as the first author or corresponding author. .
Corresponding author profile
Han Xin is a professor at the School of Integrative Medicine, School of Medicine, Nanjing University of Traditional Chinese Medicine, the head of the Department of Biochemistry and Molecular Biology and a subject leader, and a distinguished professor of Jiangsu Province. Mainly engaged in the research of microfluidic chip technology and translational medicine, gene editing and multifunctional bio-nano materials. Representative works include Science Advances, Angewandte Chemie, Small, Analytical Chemistry, Aging Cell and other domestic and foreign journals published more than 30 SCI papers.
Cha Zhengbao , Huangshan Young Scholar and Researcher, Hefei University of Technology. In recent years, aiming at the occurrence and development of microbial infections, inflammations, tumors and other diseases of body homeostasis, a series of targeted drug delivery preparations based on the physiological microenvironment of the lesion and the response of exogenous stimuli (such as light, sound, microwave, etc.) have been constructed . As the first or corresponding author, he published 44 SCI papers in Advanced Materials, Advanced Functional Materials, ACS Nano, Nano Letters, Small and other domestic and foreign journals, including 9 cover and back cover papers.
Song Yujun , Professor of the School of Modern Engineering and Applied Science, Nanjing University, PI, State Key Laboratory of Analytical Chemistry for Life. In recent years, focusing on the diagnosis and treatment of important diseases such as tumors and inflammatory bowel disease, a series of rapid diagnosis and intelligent response gene editing technology platforms based on microfluidics have been developed. He has published more than 30 SCI papers in domestic and foreign journals such as Science Advances, Nature Communications, Advanced Materials, JACS, Angewandte Chemie, Advanced Functional Materials, Nano Letters, Small, etc. as a correspondence or as the first author.
Paper information:
Controlled CRISPR-Cas9 Ribonucleoprotein Delivery for Sensitized Photothermal Therapy
Chao Chen, Yupei Ma, Shiyu Du, Yueyao Wu, Peiliang Shen, Tao Yan, Xueqing Li, Yujun Song*, Zhengbao Zha*, Xin Han*
Small
DOI: 10.1002/smll.202101155
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