Article
Chemistry, Multidisciplinary
Prithvijit Mukherjee, Cesar A. Patino, Nibir Pathak, Vincent Lemaitre, Horacio D. Espinosa
Summary: Genome engineering using CRISPR/Cas systems has provided new approaches for pharmacological screening and studying the molecular mechanisms of diseases. However, current methods for intracellular delivery and cell manipulation can be harsh, especially for sensitive cell types. In this study, an automated nanofountain probe electroporation (NFP-E) system is introduced, which offers superior cell viability and efficiency compared to traditional methods. The automated NFP-E, combined with microconfinement arrays for cell isolation, can potentially be used for CRISPR/Cas9 gene editing and cell tracking in screening studies and isogenic cell line generation.
Review
Chemistry, Multidisciplinary
Peng Yang, Athena Yue-Tung Lee, Jingjing Xue, Shih-Jie Chou, Calvin Lee, Patrick Tseng, Tiffany X. Zhang, Yazhen Zhu, Junseok Lee, Shih-Hwa Chiou, Hsian-Rong Tseng
Summary: The CRISPR/Cas9 genome editing system has revolutionized the field of therapeutic applications for genetic diseases and cancers. Efficient delivery of the system in vivo remains a significant challenge. Non-viral nano-vectors offer advantages over viral vectors and can be used to deliver CRISPR/Cas9 cargoes for targeted gene editing. The article highlights recent advances in nano-vector delivery for treating cancers and genetic diseases and proposes strategies for future development.
Article
Chemistry, Multidisciplinary
Mohammad Aghaamoo, Yu-Hsi Chen, Xuan Li, Neha Garg, Ruoyu Jiang, Jeremy Tian-Hao Yun, Abraham Phillip Lee
Summary: AESOP is a high-throughput non-viral intracellular delivery platform that allows dosage control of large cargos by optimizing delivery efficiency through mechanical shear and electric field expansion of nanopores. With acoustic microstreaming vortices, it enables efficient and uniform delivery of various molecule sizes to cells at a rate of 1 million cells per minute.
Review
Biochemistry & Molecular Biology
Xinyue Lu, Miaomiao Zhang, Ge Li, Shixin Zhang, Jingbo Zhang, Xiaoge Fu, Fengying Sun
Summary: The rapid advancements in gene therapy have opened up new possibilities for treating genetic disorders. The utilization of CRISPR/Cas9 system enables precise targeting of genes and has demonstrated remarkable efficacy. However, the development of new vectors is necessary to protect nucleic acids from degradation.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2023)
Article
Chemistry, Physical
Juhee Lee, Yoo Kyung Kang, Eonju Oh, Juhee Jeong, San Hae Im, Duk Ki Kim, Haeshin Lee, Sang-Gyu Kim, Keehoon Jung, Hyun Jung Chung
Summary: The study presents a cancer gene therapy strategy based on NanoRNP that efficiently blocks the PD-L1 immune checkpoint and induces an antitumor effect in vivo without the need for combination therapy. In vivo results demonstrate that NanoRNP can induce indels in target cells at high frequencies, significantly suppressing tumor growth.
CHEMISTRY OF MATERIALS
(2022)
Article
Multidisciplinary Sciences
Vanessa Chenouard, Isabelle Leray, Laurent Tesson, Severine Remy, Alasdair Allan, Daniel Archer, Adam Caulder, Agnes Fortun, Karine Bernardeau, Yacine Cherifi, Lydia Teboul, Laurent David, Ignacio Anegon
Summary: CRISPR-Cas9 cleavage efficacy and accuracy are influenced by the optimal formation of the ribonucleoprotein (RNP) complex. The study shows that an equimolar ratio of Cas9 and guide RNA (gRNA) is optimal for RNP complex formation. Increasing the Cas9/gRNA ratio does not improve knock-in (KI) efficiency, and excess gRNA leads to a decrease in point mutation KI efficiency and an increase in large deletions.
Article
Chemistry, Multidisciplinary
Xuan Yang, Xiaowei Wen, Jie Dai, Yanming Chen, Wanchuan Ding, Jun Wang, Xiang Gu, Xuejin Zhang, Jin Chen, Roy L. Sutliff, Steven R. Emory, Gang Ruan
Summary: This study investigates nanoparticle transport in hard-to-transfect cells using single-particle motion analysis, cell biology studies, and mathematical modeling. The researchers find significant differences in the transport of nanoparticles in bone marrow-derived mesenchymal stem cells (BMSCs) compared to HeLa cells. They also discover that BMSCs use different biological pathways for cellular uptake, vesicular transport, and exocytosis of nanoparticles compared to HeLa cells. A reaction-diffusion-advection model is used to mathematically integrate the steps of cellular transport and predict nanoparticle delivery in BMSCs. This work provides important insights into nanoparticle transport in hard-to-transfect cells and can guide the design of tailored nanoparticle-based delivery systems.
Article
Chemistry, Multidisciplinary
Xiaoyu Xu, Chang Liu, Shengyi Wang, Ermei Makila, Jiali Wang, Oliver Koivisto, Junnian Zhou, Jessica M. Rosenholm, Yilai Shu, Hongbo Zhang
Summary: In this study, a microfluidic-assisted metal-organic framework (MOF) based biomineralization strategy was developed for the efficient and remote control delivery of CRISPR/Cas9 RNP gene editing. The strategy involved growing thermo-responsive EuMOFs onto a photothermal template Prussian blue (PB) and loading RNP during MOFs crystallization in microfluidic channels. The resulting nanocarrier showed high loading efficiency, protection of RNP, and NIR-stimulated release capacity. Upon laser exposure, the nanocarrier demonstrated effective endosomal escape and precise gene knockout. The gene-editing activity could be programmed by tuning exposure times, providing a more controllable editing modality compared to the control group without laser irradiation.
Review
Pharmacology & Pharmacy
Xiaoyu Xu, Chang Liu, Yonghui Wang, Oliver Koivisto, Junnian Zhou, Yilai Shu, Hongbo Zhang
Summary: CRISPR/Cas9 is a potent gene-editing technology that shows promise for cancer therapy, but faces challenges in clinical applications. Nanotechnology plays a significant role in cancer drug delivery, aiding in overcoming the barriers of in vivo delivery for CRISPR/Cas9.
ADVANCED DRUG DELIVERY REVIEWS
(2021)
Article
Biotechnology & Applied Microbiology
Dimitra Synefiaridou, Jan-Willem Veening
Summary: CRISPR-Cas systems provide adaptive immunity for bacteria and can be used for genome editing. Researchers developed a CRISPR-Cas9 system on a plasmid for genome editing in Streptococcus pneumoniae, enabling easy and consecutive rounds of targeted gene knockout and genome editing in the opportunistic human pathogen.
APPLIED AND ENVIRONMENTAL MICROBIOLOGY
(2021)
Article
Chemistry, Multidisciplinary
Hojeong Shin, Dal-Hee Min
Summary: In this study, carbon nanodots (CNDs) were synthesized as an efficient mRNA delivery vehicle using a one-step microwave-assisted method. CNDs formed complexes with mRNA molecules through electrostatic interactions and exhibited highly effective gene delivery performance in hard-to-transfect cells. Considering their outstanding transfection ability, CNDs are expected to have further applications in mRNA-based cellular engineering.
Article
Pharmacology & Pharmacy
Jianxi Huang, Guanyou Lin, Taylor Juenke, Seokhwan Chung, Nicholas Lai, Tianxin Zhang, Tianyi Zhang, Miqin Zhang
Summary: This study introduces a novel approach using iron oxide nanoparticles for safe and efficient mRNA delivery to hard-to-transfect cancer cells. The developed nanoparticles showed enhanced cellular internalization of mRNA, easy mRNA release, and minimal cytotoxicity, suggesting great promise for mRNA therapy against hard-to-transfect cancers.
Review
Materials Science, Multidisciplinary
Shao Wei Hu, Tao Ding, Honghai Tang, Huiping Guo, Wenguo Cui, Yilai Shu
Summary: With the advancement of genome editing techniques, gene therapy is increasingly being used for the treatment of various human diseases. Nanomaterials have shown great potential in improving the efficiency and safety of gene editing in gene therapy. This review introduces three gene editing tools, with a particular focus on the development and optimization of the CRISPR system. It also discusses the recent application of nanobiomaterials in gene therapy, along with the prospects and remaining challenges. Lastly, future directions for gene editing tools, nanobiomaterial vectors, and gene therapy are presented.
Review
Biochemistry & Molecular Biology
Dengyun Nie, Ting Guo, Miao Yue, Wenya Li, Xinyu Zong, Yinxing Zhu, Junxing Huang, Mei Lin
Summary: CRISPR/Cas9 technology is a powerful gene editing tool with great potential in oncology. However, its efficiency and safety issues restrict its widespread application. Developing a suitable delivery method, such as nanoparticles, is urgently needed.
Article
Biotechnology & Applied Microbiology
Mei Zhao, Yi-Dan Sun, Mengdi Yin, Juan-Juan Zhao, Si-Ang Li, Guohua Li, Feng Zhang, Jing Xu, Fei-Ying Meng, Beldon Zhang, Xin-Yu Sun, Jian-Ping Zhang, Tao Cheng, Xiao-Bing Zhang
Summary: This study successfully treated hemophilia A in a mouse model using a gene-editing strategy and identified the humoral immune response as the main cause of decreased treatment efficacy. The findings highlight the importance of modulating the innate immune response triggered by liver damage.
HUMAN GENE THERAPY
(2022)
Article
Biochemistry & Molecular Biology
N. Balneger, L. A. M. Cornelissen, M. Wassink, S. J. Moons, T. J. Boltje, Y. E. Bar-Ephraim, K. K. Das, J. N. Sondergaard, C. Bull, G. J. Adema
Summary: The study reveals that sialic acids play a crucial role in regulating the activation and interaction of dendritic cells. Blocking sialic acids enhances the immune activation ability and antigen-induced T cell interactions of dendritic cells. These findings have important implications for understanding the mechanisms of immune responses.
CELLULAR AND MOLECULAR LIFE SCIENCES
(2022)
Article
Biochemistry & Molecular Biology
Sandhya Malla, Devi Prasad Bhattarai, Paula Groza, Dario Melguizo-Sanchis, Ionut Atanasoai, Carlos Martinez-Gamero, Angel-Carlos Roman, Dandan Zhu, Dung-Fang Lee, Claudia Kutter, Francesca Aguilo
Summary: This study reveals the role of ZFP207 in mouse embryonic stem cells through multiple functional analyses. Unlike human ESCs, mouse ZFP207 mainly exerts its effects by controlling AS networks and acting as RNA-binding proteins (RBPs) to maintain ESC identity and regulate neural fate commitment.
Article
Biochemistry & Molecular Biology
William Gao, Carlos J. Gallardo-Dodd, Claudia Kutter
Summary: This study examines the correlation between codon and anticodon pools and their impact on translation efficiency at the individual cell level. The researchers found that codon usage and amino acid demand are highly stable across different cell types. They also discovered a strong correlation between amino acid supply and demand in almost all cell types. Notably, neurons showed enhanced translation efficiency due to an increased supply of specific tRNA anticodons. This study provides valuable insights into translation elongation in mammalian cells and suggests potential implications for neurological pathologies.
Article
Gastroenterology & Hepatology
Jonas Norskov Sondergaard, Christian Sommerauer, Ionut Atanasoai, Laura C. Hinte, Keyi Geng, Giulia Guiducci, Lars Brautigam, Myriam Aouadi, Lovorka Stojic, Isabel Barragan, Claudia Kutter
Summary: By globally characterizing RNA-binding proteins (RBPs), we revealed the perturbation of RBP gene expression in hepatocellular carcinoma (HCC) and identified their additional functions in tumor growth regulation, particularly in lipid metabolism. Furthermore, the interaction between non-canonical RBP CCT3 and LINC00326 was found to be involved in the regulation of lipid accumulation and degradation, affecting HCC cell phenotype and tumour growth in vivo.
Review
Cell Biology
Christian Sommerauer, Claudia Kutter
Summary: The liver plays important roles in detoxification, energy metabolism, and whole body homeostasis. However, chronic overload of fatty acids and glucose can lead to the development of malignant phenotypes in the liver. The global increase in metabolic dysfunction-associated fatty liver disease (MAFLD) is affecting a significant portion of the population. Noncoding RNAs, including long and short noncoding RNAs, have emerged as potential targets for treatment interventions. The interactions between noncoding RNAs and proteins offer precise ways to target specific cellular pathways.
AMERICAN JOURNAL OF PHYSIOLOGY-CELL PHYSIOLOGY
(2022)
Article
Immunology
Thuy T. Luu, Jonas Norskov Sondergaard, Lucia Pena-Perez, Shabnam Kharazi, Aleksandra Krstic, Stephan Meinke, Laurent Schmied, Nicolai Frengen, Yaser Heshmati, Marcin Kierczak, Thibault Bouderlique, Arnika Kathleen Wagner, Charlotte Gustafsson, Benedict J. Chambers, Adnane Achour, Claudia Kutter, Petter Hoglund, Robert Mansson, Nadir Kadri
Summary: FOXO1 and FOXO3 coordinately regulate essential developmental genes at multiple stages during murine NK cell and ILC lineage commitment through orchestrating distinct molecular mechanisms.
FRONTIERS IN IMMUNOLOGY
(2022)
Article
Biochemistry & Molecular Biology
Keyi Geng, Lara G. Merino, Linda Wedemann, Aniek Martens, Malgorzata Sobota, Yerma P. Sanchez, Jonas Norskov Sondergaard, Robert J. White, Claudia Kutter
Summary: This study investigates the genomic rearrangements induced by the CRISPR-Cas9 system using an innovative droplet-based target enrichment approach and long-read sequencing. The researchers found extensive genomic disruptions, including genomic duplications, inversions of the target region, integrations of exogenous DNA, and rearrangements of interchromosomal DNA fragments. Furthermore, these genomic alterations were found to lead to functional aberrant DNA fragments and alter cell proliferation. The findings broaden our understanding of the consequences of the Cas9 deletion system and emphasize the importance of meticulous genomic validations.
Article
Medicine, Research & Experimental
Yuta Yamaguchi, Yasuhiro Kato, Ryuya Edahiro, Jonas N. Sondergaard, Teruaki Murakami, Saori Amiya, Shinichiro Nameki, Yuko Yoshimine, Takayoshi Morita, Yusuke Takeshima, Shuhei Sakakibara, Yoko Naito, Daisuke Motooka, Yu-Chen Liu, Yuya Shirai, Yasutaka Okita, Jun Fujimoto, Haruhiko Hirata, Yoshito Takeda, James B. Wing, Daisuke Okuzaki, Yukinori Okada, Atsushi Kumanogch
Summary: Consecutive mRNA vaccinations against SARS-CoV-2 enhance innate and adaptive immune responses. This study investigates the role of epigenetic regulation in mediating enhanced innate immune responses and the persistence of these effects. The results show that BNT162b2 mRNA vaccination upregulates antiviral and IFN-stimulated gene expression in monocytes, with greater effects after the second vaccination. However, these effects are transient and disappear 4 weeks after the second vaccination. This study provides novel insights into innate immune memory and its implications for vaccine development.
Article
Multidisciplinary Sciences
Jonas Norskov Sondergaard, Janyerkye Tulyeu, Ryuya Edahiro, Yuya Shirai, Yuta Yamaguchi, Teruaki Murakami, Takayoshi Morita, Yasuhiro Kato, Haruhiko Hirata, Yoshito Takeda, Daisuke Okuzaki, Shimon Sakaguchi, Atsushi Kumanogoh, Yukinori Okada, James Badger
Summary: Sex-specific differences in humoral immune responses to COVID-19 have been observed, with a higher loss of circulating follicular regulatory T cells (cTfr) in male patients. A network of cell types associated with extrafollicular antibody production is stronger in males and positively correlated with neutralizing antibody concentrations, while negatively correlated with cTfr frequency. These findings suggest that the balance of cTfr and the network of antibody-producing cell types may be a key factor in the altered humoral immune responses between male and female COVID-19 patients.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2023)
Article
Multidisciplinary Sciences
Takehito Yokoi, Mari Murakami, Takako Kihara, Shigeto Seno, Mitsuru Arase, James Badger Wing, Jonas Norskov Sondergaard, Ryuichi Kuwahara, Tomohiro Minagawa, Eri Oguro-Igashira, Daisuke Motooka, Daisuke Okuzaki, Ryota Mori, Atsuyo Ikeda, Yuki Sekido, Takahiro Amano, Hideki Iijima, Keiichi Ozono, Tsunekazu Mizushima, Seiichi Hirota, Hiroki Ikeuchi, Kiyoshi Takeda
Summary: This study provides insights into the pathogenesis of inflammatory bowel disease (IBD) by profiling the T cells in the intestinal mucosa. A CD4+ tissue-resident memory T cell (Trm) subset was identified, which is increased in Crohn's disease (CD) and exhibits unique inflammatory properties. Specifically, a CD-specific CD4+ Trm subset was found to be the predominant producer of type 1 inflammatory cytokines. These findings contribute to the understanding of the molecular mechanisms underlying IBD.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2022)
Article
Biochemistry & Molecular Biology
Laurent Schmied, Thuy T. Luu, Jonas N. Sondergaard, Sophia H. Hald, Stephan Meinke, Dara K. Mohammad, Sunitha B. Singh, Corinna Mayer, Giovanna Perinetti Casoni, Michael Chrobok, Heinrich Schlums, Giorgia Rota, Hieu M. Truong, Lisa S. Westerberg, Greta Guarda, Evren Alici, Arnika K. Wagner, Nadir Kadri, Yenan T. Bryceson, Mezida B. Saeed, Petter Hoglund
Summary: Natural killer (NK) cells recognize infected cells and tumors. The function of NK cells depends on balanced signaling from activating and inhibitory receptors. The subcellular localization of tyrosine phosphatase SHP-1 determines NK cell tolerance and education. Education of NK cells leads to reduced synapse accumulation of SHP-1 and augmented signaling from activating receptors.
Article
Multidisciplinary Sciences
Carlos J. Gallardo-Dodd, Christian Oertlin, Julien Record, Romulo G. Galvani, Christian Sommerauer, Nikolai V. Kuznetsov, Evangelos Doukoumopoulos, Liaqat Ali, Mariana M. S. Oliveira, Christina Seitz, Mathias Percipalle, Tijana Nikic, Anastasia A. Sadova, Sofia M. Shulgina, Vjacheslav A. Shmarov, Olga V. Kutko, Daria D. Vlasova, Kseniya D. Orlova, Marina P. Rykova, John Andersson, Piergiorgio Percipalle, Claudia Kutter, Sergey A. Ponomarev, Lisa S. Westerberg
Summary: To ensure safe space missions, understanding the impact of space flight on the immune system is crucial. A study showed that T cells undergo transcriptional alterations after 21 days of dry immersion exposure, with the most significant changes occurring after 14 days. T cells exhibited adaptive responses with a transcriptional profile similar to pre-exposure levels after 21 days, but reverted back to a different profile 7 days after the end of the immersion. These findings suggest that T cells adapt by rewiring their transcriptomes in response to simulated weightlessness and retain remodeling cues when returning to normal gravity.