Innovative method for in-cell gene manipulation

A cross-disciplinary Small Medicine and Advanced Research Translation (SMART) team led by Prof. Dar-Bin Shieh of Institute of Oral Medicine in Medical College at National Cheng Kung University (NCKU), Tainan, Taiwan, has announced a breakthrough in the precision in-cell gene scission at pre-designed sequence sites using Artificial Targeting Light Activated Nano Scissors (ATLANS) and a custom build photonic device.

The research achievement is supported under National Nano Science and Technology Program funded by National Science Council (NSC), Taiwan. This innovative discovery is recently accepted by internationally renowned journal Biomaterials and is currently under patent application.

The nano-enabled novel technology recognize, capture and perform double strand cutting of the desired DNA sequence like restriction endonuclease did but it is completely artificially synthesized and controlled by specific photon energy. In addition, the team successfully shut down target drug resistant gene STAT3 in cancer cell using ATLANS and created a new inspiration toward future cancer gene therapy.

NCKU President Michael Ming-Chiao Lai expressed in the press conference of the ATLANS, "There are many technical difficulties of making modifications to cell genes and treating various diseases. In the past, even though we can enter cells to destroy genes with virus or drugs, we cannot control the genes properly. The ATLANS developed by the research team led by Prof. Dar-Bin Shieh is an innovative method which can target specific genes in the cells and make modifications, thus it is a very significant breakthrough of the clinical research technology."

Prof. Dar-Bin Shieh pointed out, "Many human diseases such as cancer or genetic diseases are caused by gene mutations. Cancer cell and infectious pathogen may develop drug resistance through activation of certain genes. Therefore, one of the major values of the Artificial Targeting Light Activated Nano Scissor is its ability to identify the target genes in the nucleus and optically-control the precise genetic cutting to block its pathogenic functions, thus reaching treatment effect."

Prof. Dar-Bin Shieh explained, "The ATLANS is an innovative method and it is also the first time this method is applied for in-cell gene manipulation. This technology uses nanoparticles as a quencher to protect the cutter from non-specific activation at wrong sequence sites and also protect the entire TFO layer from being disrupted by the body before reaching the final destination. Once the genetic targets are locked on, the ATLANS will be activated as the proximity of the photo-cutter is no longer restricted by the particle surface plasma. This precision molecular dynamic control is required during the execution of gene cutting in the atomic level accuracy like a molecular 'Nano Scissors.'"

With the Nano Scissors technology, Prof. Dar-Bin Shieh's research team further developed a GeneErasor optical system, which functions to "erase" specific target gene in the culture cell. The gene eraser is anticipated to test-run early next year. The GeneErasor system is currently of blue-ray band. The team has successfully tested a second-generation system in test tube for near-infrared laser-scan activation that performs better tissue penetration for diseases in deep organ system.

Cross-disciplinary medical research has become a global trend. The innovations of the SMART research team led by Prof. Dar-Bin Shieh has set an excellent model of trans-disciplinary and cross-university clinical-oriented research integration and demonstrated advanced niches for medical innovations, integrating the knowledge and experience of organic chemistry, optoelectronic, nano-synthesis, biochemistry, molecular biology and genomic medicine scholars and experts and creating great achievements under the long-term support of Taiwan National Science and Technology Program for Nanoscience and Nanotechnology, National Science Council, Taiwan.

Prof. Dar-Bin Shieh believed, "The success of the research achievement is due to the joint effort of the team. National Cheng Kung University has provided an excellent environment, allowing professors with different disciplines from College of Medicine, Center of Excellent for Clinical Trials and Research in Oncology Specialty, College of Electrical Engineering and Computer Science, College of Engineering, and College of Science to form a research team, find effective solutions to clinical issues and benefit future patients through the combination of medicine and clinic. The University is proposing a new concept to foster clinical and industrial translation of fundamental discovery and engineering power through a partnership network, the Center for Biomedical Excellent and Synergy in Taiwan (The BEST Center)."

Director Wu-Chou Su of Center of Excellent for Clinical Trials and Research in Oncology Specialty is an outstanding clinical oncologist. He stated, "All cancer diseases arise from genetic mutations. The Artificial Targeting Light Activated Nano Scissor applied on cancer treatment initially focuses on late stage. We still need to put in more efforts to package ATLANS for effective tumor-targeting when entering human body. The ATLANS based therapy to modify tumor sensitivity to anti-cancer drugs and improve early disease intervention is still a challenging yet to be confronted."

The members of the nanomedical research team led by Prof. Dar-Bin Shieh of Institute of Oral Medicine in NCKU Medical College include Director Wu-Chou Su of NCKU Center of Excellent for Clinical Trials and Research in Oncology Specialty; Dean Yonghua Tommy Tzeng of College of Electrical Engineering and Computer Science; Distinguished Professor Chen-Sheng Yeh of Chemical Department; and Professor Jih Ru Hwu of National Tsing Hua University Chemistry Department who synthesized the photocleaver compound. The doctoral student Tsung-Lin Tsai of NCKU Basic Medical Science Institute and the master student Tsung-Ju Li of NCKU Institute of Oral Medicine have put in significant efforts in this work.

Through the cooperation with Dr. Wah Chiu, the Director of National Center for Macromolecular Imaging at Baylor University in United States, the team has preliminarily analyzed the ultrastructure of ATLANS and begun the development of ATALANS-2 and also the advanced nonlinear optical system GeneErasor-3D, hoping to achieve major breakthroughs in the future.