Article
Biotechnology & Applied Microbiology
Samaneh Ghanbari, Elham Bayat, Masoumeh Azizi, Pezhman Fard-Esfahani, Mohammad Hossein Modarressi, Fatemeh Davami
Summary: Recombinant CHO cell line development traditionally relies on random integration (RI) approach, but targeted integration offers potential advantages in terms of control and stability. In this study, we used the CRISPR-mediated precise integration into target chromosome (CRIS-PITCh) system combined with the Bxb1 recombinase-mediated cassette exchange (RMCE) system to generate a transgene-expressing cell line with improved specificity and stability.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2023)
Article
Cell & Tissue Engineering
Huan Zou, Mingfeng Guan, Yundong Li, Fang Luo, Wenyuan Wang, Yiren Qin
Summary: This study successfully corrected the genetic mutation of Achondroplasia patients using stem cells and gene editing technology, restoring their chondrogenic differentiation ability and providing an important theoretical and experimental basis for Achondroplasia research and treatment.
STEM CELL RESEARCH & THERAPY
(2021)
Article
Biochemical Research Methods
Jin Myeong Kwak, Youngsik Lee, Sung Wook Shin, Jae Seong Lee
Summary: A cell cycle enrichment method using a lower concentration of hydroxyurea (HU) has been developed to increase HDR-mediated KI efficiency in CHO cells, resulting in a 1.2-1.5 fold improvement in KI efficiency compared to control conditions.
Article
Biotechnology & Applied Microbiology
James S. Donaldson, Matthew P. Dale, Susan J. Rosser
Summary: By decoupling cell growth and biotherapeutic production, it is possible to enhance production efficiency and improve cell growth characteristics, particularly for difficult-to-express proteins.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2021)
Article
Biotechnology & Applied Microbiology
Rajveer Singh, Shivani Chandel, Arijit Ghosh, Anupam Gautam, Daniel H. Huson, V. Ravichandiran, Dipanjan Ghosh
Summary: In the past two decades, yeast has been widely used as a cost-effective biological tool for producing small molecules and biofuels. The application of CRISPR-Cas techniques in yeast genetic and metabolic engineering has revolutionized the field, particularly in targeted chromosomal integration using synthetic donor constructs. This study presents a highly efficient strategy for targeted chromosomal integration and in-frame expression of foreign genes in the yeast genome using CRISPR-Cas9 and homology-dependent recombination (HDR). The optimized methods achieved a 74% efficiency in integrating small constructs at multiple target sites. The study also demonstrated that as few as 15 base pairs of microhomology are sufficient for targeted knock-in with minimal construct concentration. Whole-genome sequencing confirmed the absence of off-target effects. This protocol provides a comprehensive and streamlined approach for integrating essential genes into the yeast genome for synthetic biology and industrial purposes.
Article
Multidisciplinary Sciences
Kathryn Rozen-Gagnon, Soon Yi, Eliana Jacobson, Sasha Novack, Charles M. Rice
Summary: This study developed a mosquito-optimized, plasmid-based CRISPR/Cas9 system for use in U4.4 (Ae. albopictus) and Aag2 (Ae. aegypti) cell lines, demonstrating highly efficient editing of the AGO1 locus. The experimentally verified plasmids are versatile, cost-effective, and efficiently edit immune competent mosquito cell lines commonly used in arbovirus studies, enabling knock-in of a 3xFLAG tag at the N-terminus of endogenous AGO1.
SCIENTIFIC REPORTS
(2021)
Article
Biochemistry & Molecular Biology
Chengpeng Wang, Yang Li, Na Wang, Qin Yu, Yonghong Li, Junping Gao, Xiaofeng Zhou, Nan Ma
Summary: We established a platform for CRISPR/Cas9-mediated gene editing in rose using suspension cells, and successfully edited a gene involved in the ethylene signaling pathway. Our optimized CRISPR/Cas9 system provides a powerful tool for functional genomics, molecular breeding, and synthetic biology in rose.
JOURNAL OF INTEGRATIVE PLANT BIOLOGY
(2023)
Article
Chemistry, Multidisciplinary
Weimin Ruan, Mingzhu Jiao, Sen Xu, Muhammad Ismail, Xuan Xie, Yang An, Haixing Guo, Rongjun Qian, Bingyang Shi, Meng Zheng
Summary: The brain-targeted CRISPR/Cas9 based nanomedicine successfully crossed the blood-brain barrier and accumulated in brain tumors with the help of angiopep-2 peptide functionality, leading to an improved gene knockout efficiency for glioblastoma treatment and suppression of tumor growth.
JOURNAL OF CONTROLLED RELEASE
(2022)
Article
Cell Biology
Binbin Ying, Yoshinori Kawabe, Feiyang Zheng, Yuki Amamoto, Masamichi Kamihira
Summary: This study aimed to develop a high-production system using a synthetic biology approach, by designing a gene expression system based on an artificial transcription factor to strongly induce the high expression of target genes in CHO cells, for the purpose of high-yield antibody production.
Article
Plant Sciences
Yudong Chen, Haiying Xiang, Ling Jia, Qianxu Yang, Jianduo Zhang, Jiarui Jiang, Wanli Zeng, Lele Deng, Jingjing Jin, Qian Gao, Xuemei Li
Summary: A total of 4382 sgRNAs targeting 1060 tobacco genes were obtained, resulting in the creation of 10,682 targeted mutants using high-throughput methods. Four optimization experiments were conducted to address issues encountered during the genetic transformation process.
PLANT CELL REPORTS
(2023)
Review
Medicine, Research & Experimental
Asma Ghaemi, Elnaz Bagheri, Khalil Abnous, Seyed Mohammad Taghdisi, Mohammad Ramezani, Mona Alibolandi
Summary: The prokaryotic CRISPR-Cas systems serve as a revolutionary genome editing tool for modifying, visualizing, and identifying DNA and RNA sequences in live cells of various species. Besides genome editing, the DNA-targeting modifications by CRISPR-Cas offer opportunities for diagnosis, therapy, and genetic disorders investigation. Preclinical studies and CRISPR-Cas9-based therapeutics may encounter challenges, but there is potential for its development in cancer therapy.
Article
Plant Sciences
Meng Li, Xufang Niu, Shuang Li, Shasha Fu, Qianfang Li, Meizhi Xu, Chunhua Wang, Shuang Wu
Summary: CRISPR/Cas9-mediated gene knockout is an important tool for studying gene function. Engineering the Cas9 system to achieve cell-type-specific knockout of functional genes is useful for understanding the specific functions of genes in different cell types.
Article
Chemistry, Multidisciplinary
Yuyuan Wang, Pawan K. Shahi, Xiuxiu Wang, Ruosen Xie, Yi Zhao, Min Wu, Seth Roge, Bikash R. Pattnaik, Shaoqin Gong
Summary: A stimulus-responsive silica nanoparticle has been developed for efficient delivery of nucleic acids and CRISPR genome editors, showing great potential for applications in gene therapy and genome editing.
JOURNAL OF CONTROLLED RELEASE
(2021)
Article
Plant Sciences
Jun You, Donghua Li, Li Yang, Senouwa Segla Koffi Dossou, Rong Zhou, Yanxin Zhang, Linhai Wang
Summary: The CRISPR/Cas9 system was successfully applied to sesame to induce targeted mutagenesis in specific genes. The results demonstrate the efficiency and effectiveness of CRISPR/Cas9 coupled with hairy root transformation in assessing gene functions in sesame. This study provides important insights into gene functional research and molecular breeding in sesame.
FRONTIERS IN PLANT SCIENCE
(2022)
Article
Genetics & Heredity
Zhenying Liu, Mingjun Zhang, Peixuan Huang, Zhaoyang Ji, Chunyun Qi, Shuyu Jiao, Dehua Zhao, Yuan Jiang, Xue Chen, Dongmei Lv, Daxin Pang, Xin Zhang, Li Feng, Zicong Xie, Hongsheng Ouyang
Summary: This study optimized the gene editing strategy and successfully generated APN-chimeric pigs with normal reproductive abilities and the ability to effectively inhibit TGEV infection, providing a valuable tool for studying porcine enteric coronaviruses and developing antiviral solutions.
Article
Biotechnology & Applied Microbiology
Kenji Masuda, Kazuhiko Watanabe, Tomonori Ueno, Yuto Nakazawa, Yumiko Tanabe, Yuko Ushiki-Kaku, Kiyoko Ogawa-Goto, Yukikazu Ehara, Hisashi Saeki, Takeshi Okumura, Koichi Nonaka, Masamichi Kamihira
Summary: Researchers attempted to apply translational enhancing technology to high antibody-producing cell lines, and by overexpressing key genes p180 and SF3b4, they increased the antibody yield and specific productivity. The results indicate that the overexpression of p180 and SF3b4 is a promising approach for establishing high-producer cell lines.
JOURNAL OF BIOSCIENCE AND BIOENGINEERING
(2022)
Article
Materials Science, Multidisciplinary
Xiaoting Fu, Seiichi Ohta, Takahiro Kawakatsu, Masamichi Kamihira, Yasuyuki Sakai, Taichi Ito
Summary: Inspired by the unique structure of human red blood cells, novel deformable core-shell particles cDFCs have been developed as oxygen carriers. These particles have shown efficiency in delivering oxygen to multiple layered cells under hypoxic conditions, indicating their potential application in tissue engineering and blood substitution in the future.
ADVANCED MATERIALS TECHNOLOGIES
(2022)
Article
Engineering, Marine
Dipak Pandey, Takahiro Matsubara, Taiju Saito, Yukinori Kazeto, Koichiro Gen, Tetsushi Sakuma, Takashi Yamamoto, Miyuki Mekuchi, Rie Goto
Summary: The study utilized TALEN technology to achieve gene knockout in the kawakawa slc24a5 gene, investigating gene expression and functional differences, revealing association with melanin pigment formation. Experimental results showed that most TALEN-induced mutants exhibited mosaic patterns in retinal pigment epithelium cells.
JOURNAL OF MARINE SCIENCE AND ENGINEERING
(2021)
Article
Cell Biology
Kaichi Watanabe, Yuhei Yasui, Yuta Kurose, Masashi Fujii, Takashi Yamamoto, Naoaki Sakamoto, Akinori Awazu
Summary: This study investigated the process of gastrulation in sea urchin embryos and found that omeprazole may induce partial exogastrulation by disrupting the apical-basal polarity of F-actin distribution.
Article
Multidisciplinary Sciences
Yoshinori Kawabe, Masamichi Kamihira
Summary: This study analyzed the phenotypic changes, gene expression profiles, and karyotype of immortalized kidney cells (CHK-Q) derived from Chinese hamster tissue. The results suggest that CHK-Q cells may have transformed from renal stem cells and the immortalization process is associated with increased fluctuations in the expression of specific proto-oncogenes.
Article
Cell Biology
Hiroyuki Kitano, Yoshinori Kawabe, Masamichi Kamihira
Summary: Researchers have developed a human hepatoma cell line with heat-inducible liver functions, which can be easily prepared in large quantities. These genetically modified cells demonstrate enhanced liver functions after heat treatment, and further enhancements can be seen in three-dimensional spheroid cultures. Comprehensive transcriptome analysis revealed increased expression of liver function-related genes in these cells after heat treatment.
Article
Oncology
Masahiro Kaneko, Akio Yamaguchi, Akira Ito
Summary: This study demonstrates that the redox phospholipid polymer pMFc induces immunogenic cell death (ICD) in murine colon cancer cells and elicits antitumor immunity. pMFc produces oxidative stress by extracting electrons from cancer cells and releases DAMPs to induce ICD.
Article
Biotechnology & Applied Microbiology
Taisei Wakabayashi, Masahiro Kaneko, Tomoki Nakai, Masanobu Horie, Hiroyuki Fujimoto, Masazumi Takahashi, Shota Tanoue, Akira Ito
Summary: This study describes a nanowarming technology that successfully rewarms vitrified pancreatic islet cells and improves their viability. This technology has the potential to solve the issue of transplantation in pancreatic islet cell biobanking.
BIOENGINEERING & TRANSLATIONAL MEDICINE
(2023)
Article
Oncology
Ami Nishikawa, Yutaro Suzuki, Masahiro Kaneko, Akira Ito
Summary: Hyperthermia using magnetic nanoparticles can destroy tumor tissues and induce inflammation. However, this approach is insufficient for poorly immunogenic tumors. This study found that combining an HMGB1 inhibitor with immunomodulators can achieve complete tumor regression.
CANCER IMMUNOLOGY IMMUNOTHERAPY
(2023)
Article
Gastroenterology & Hepatology
Tomoki Takashina, Akihiro Matsunaga, Yukiko Shimizu, Tetsushi Sakuma, Tadashi Okamura, Kunie Matsuoka, Takashi Yamamoto, Yukihito Ishizaka
Summary: A protein-based artificial transcription system was developed to engineer hepatocyte-like cells from human umbilical cord-derived mesenchymal stem cells. The engineered cells exhibited functional and molecular characteristics similar to hepatocytes, and significantly improved the survival rate in mice with hepatic failure. This transcription system could serve as a versatile tool for cell therapy for hepatic failures.
HEPATOLOGY COMMUNICATIONS
(2023)
Article
Oncology
Masahiro Kaneko, Hiroto Yamazaki, Takahiro Ono, Masanobu Horie, Akira Ito
Summary: Magnetic hyperthermia (MHT) is a promising cancer treatment that uses the heat generated by nano-heaters (such as magnetite nanoparticles) under an alternating magnetic field to specifically damage tumor tissue. Researchers have improved the therapeutic efficacy of MHT by using mitochondria-targeting nanoparticles, which have shown enhanced therapeutic effects.
Article
Biotechnology & Applied Microbiology
Masahiro Kaneko, Hiroaki Moriguchi, Ryo Futatsubashi, Satoru Ayano, Goro Kobayashi, Akira Ito
Summary: In this study, a new encapsulation device was developed to encapsulate pancreatic beta cells in a semipermeable membrane bag. The device demonstrated insulin secretion and blood glucose reduction in vitro and in diabetic mice, without triggering immune responses.
JOURNAL OF BIOSCIENCE AND BIOENGINEERING
(2023)
Article
Biotechnology & Applied Microbiology
Masahiro Kaneko, Airi Sato, Satoru Ayano, Akio Fujita, Goro Kobayashi, Akira Ito
Summary: This study reports a novel poly(vinyl alcohol) microcarrier for the expansion of hMSCs in suspension culture, and demonstrates long-term expansion using a bead-to-bead cell transfer method. The study also presents magnetic PVA microcarriers for cell recovery.
JOURNAL OF BIOSCIENCE AND BIOENGINEERING
(2023)
Article
Cell Biology
Binbin Ying, Yoshinori Kawabe, Feiyang Zheng, Yuki Amamoto, Masamichi Kamihira
Summary: This study aimed to develop a high-production system using a synthetic biology approach, by designing a gene expression system based on an artificial transcription factor to strongly induce the high expression of target genes in CHO cells, for the purpose of high-yield antibody production.
Article
Polymer Science
Yusuke Sakai, Yoshihiro Kubo, Nana Shirakigawa, Yoshinori Kawabe, Masamichi Kamihira, Hiroyuki Ijima
Summary: Researchers have developed a technique to rapidly form kidney-like tissues (cysts) in vitro by combining an organ-derived extracellular matrix gel culture system and a renal stem cell line. The cysts formed from the renal stem cells fused together and expanded while maintaining three-dimensional structures. The expression of genes related to kidney development and maturation increased compared to traditional monolayer cultures.
