Article
Engineering, Biomedical
Yuqian Jiang, Rachel Catherine Hoenisch, Yun Chang, Xiaoping Bao, Craig E. Cameron, Xiaojun Lance Lian
Summary: In this study, two all-in-one vectors were developed for efficient delivery of CRISPR components, achieving robust genome editing and RNA interference. Utilizing the PiggyBac transposon system, these vectors exhibited high efficiency in inducing gene knockout and RNA knockdown in human pluripotent stem cells (hPSCs).
BIOACTIVE MATERIALS
(2022)
Article
Biotechnology & Applied Microbiology
Ayako Nishizawa-Yokoi, Seiichi Toki
Summary: The study presents a piggyBac-mediated transgenesis system for temporary expression of CRISPR/Cas9 in plants, enabling precise excision of the piggyBac after successful targeted mutagenesis induction. The system demonstrated successful transgenesis in rice plants and the detection of CRISPR/Cas9-induced mutations even after piggyBac excision, suggesting its potential as an efficient tool for CRISPR/Cas9-mediated targeted mutagenesis in vegetatively propagated crops.
PLANT BIOTECHNOLOGY JOURNAL
(2021)
Review
Genetics & Heredity
Peter J. Chen, David R. Liu
Summary: In this review, the authors discuss the latest developments in prime editing systems, highlighting improvements in editing efficiency and capabilities, as well as emerging applications in research and preclinical therapeutic studies. Prime editors offer versatility, specificity, and precision in making targeted DNA changes in living cells. The review summarizes prime editing strategies, addresses their limitations, discusses recent developments to overcome these limitations, and explores potential applications and future directions.
NATURE REVIEWS GENETICS
(2023)
Review
Biochemistry & Molecular Biology
Chenyu Lu, Jingyu Kuang, Tong Shao, Sisi Xie, Ming Li, Lingyun Zhu, Lvyun Zhu
Summary: Prime editing technology has shown attractive potential for versatile genome editing ability, but improvements are needed in terms of editing efficiency, off-target effects, and delivery systems. Recent optimizations have resulted in promising prospects for its application in various fields. This review provides a concise insight into the development, barriers, optimization efforts, and application directions of prime editing, offering clues for future advancements in this emerging field.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Multidisciplinary Sciences
Justin A. Bosch, Gabriel Birchak, Norbert Perrimon
Summary: The study successfully applied prime editing in Drosophila to introduce premature stop codons and achieve efficient germline transmission, demonstrating its potential for studying gene function in Drosophila.
PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
(2021)
Review
Biotechnology & Applied Microbiology
Zhangrao Huang, Gang Liu
Summary: Prime editing is a precise genome manipulation technology that utilizes the CRISPR-Cas9 system's search and replace approach, without the need for exogenous donor DNA and DNA double-strand breaks (DSBs). It offers a wider editing scope compared to base editing. Prime editing has shown promise in breeding and genomic functional studies of animals and plants, disease treatment, and modification of microbial strains, as it has been successfully applied in various plant cells, animal cells, and Escherichia coli. This paper provides a brief overview of the basic strategies of prime editing, summarizes its research progress and prospects in multiple species, and outlines various optimization strategies to improve its efficiency and specificity.
FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY
(2023)
Article
Biotechnology & Applied Microbiology
Hui Kwon Kim, Goosang Yu, Jinman Park, Seonwoo Min, Sungtae Lee, Sungroh Yoon, Hyongbum Henry Kim
Summary: This study identified factors affecting PE2 efficiency through high-throughput evaluation and developed three computational models to predict pegRNA efficiency, which can be applied to edits of various types and positions. Spearman's correlations between 0.47 and 0.81 were found when testing the accuracy of the predictions using independent test data sets.
NATURE BIOTECHNOLOGY
(2021)
Review
Biotechnology & Applied Microbiology
Gregory A. Newby, David R. Liu
Summary: Recent advances in genome editing technologies have made single-dose cures for genetic diseases more promising. Base editors and prime editors offer greater precision and efficiency compared to nuclease-dependent editing approaches, with less unwanted byproducts. Delivery of these editors in animal models has shown promise for in vivo therapeutic genome editing.
Article
Biochemical Research Methods
Kylie Standage-Beier, Stefan J. Tekel, David A. Brafman, Xiao Wang
Summary: This study introduces a software tool called PINE-CONE which enables high-throughput automated design of pegRNAs and prime editing strategies. By translating edit coordinates and sequences into required designs, PINE-CONE accelerates the application of PE technology in synthetic biology and biomedical research.
ACS SYNTHETIC BIOLOGY
(2021)
Article
Biotechnology & Applied Microbiology
Karl Petri, Weiting Zhang, Junyan Ma, Andrea Schmidts, Hyunho Lee, Joy E. Horng, Daniel Y. Kim, Ibrahim C. Kurt, Kendell Clement, Jonathan Y. Hsu, Luca Pinello, Marcela Maus, J. Keith Joung, Jing-Ruey Joanna Yeh
Summary: This study demonstrated prime editing using purified ribonucleoprotein complexes, achieving desired edits in zebrafish embryos and human cells. However, unintended insertions, deletions, and guide RNA scaffold incorporations were also observed.
NATURE BIOTECHNOLOGY
(2022)
Review
Biochemistry & Molecular Biology
Jingying Li, Chen Zhang, Yubing He, Shaoya Li, Lei Yan, Yucai Li, Ziwei Zhu, Lanqin Xia
Summary: Precise replacement of an allele with an elite allele controlling an important agronomic trait in a predefined manner by gene editing technologies is highly desirable in crop improvement. Base editing and prime editing are two newly developed precision gene editing systems which can introduce the substitution of a single base and install the desired short indels to the target loci in the absence of double-strand breaks and donor repair templates, respectively. Various strategies have been attempted to optimize both base editor (BE) and prime editor (PE) to improve their editing efficacy, specificity, and expand the targeting scopes. Based on the latest development, appropriate BEs and PEs are recommended for basic plant research and crop improvement, and further optimization of these editors is proposed. Both BEs and PEs are envisioned to become routine and customized precise gene editing tools for plant biological research and crop improvement in the near future.
JOURNAL OF INTEGRATIVE PLANT BIOLOGY
(2023)
Article
Biochemical Research Methods
Lijun Hao, Xiangdong Pu, Jingyuan Song
Summary: Prime editing, a new gene editing technology based on CRISPR/Cas, allows for direct and precise editing of specified DNA sites without double strand breaks and donor DNA, offering a wider range of editing types and potential editing capabilities compared to traditional CRISPR/Cas9 and base editing. Successfully developed in mammalian cells and applied in plants, prime editing has the advantage of direct DNA editing without causing double strand breaks, providing a new approach for genetic improvement in plants.
Article
Biochemistry & Molecular Biology
Dexin Qiao, Junya Wang, Min-Hui Lu, Cuiping Xin, Yiping Chai, Yuanyuan Jiang, Wei Sun, Zhenghong Cao, Siyi Guo, Xue-Chen Wang, Qi-Jun Chen
Summary: This study tested PEs optimized through three strategies in maize and demonstrated that the ePE5max system efficiently generated heritable mutations that conferred resistance to herbicides in maize target genes.
JOURNAL OF INTEGRATIVE PLANT BIOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Ming Lyu, Yongsen Sun, Nana Yan, Qiang Chen, Xin Wang, Zehui Wei, Zhiying Zhang, Kun Xu
Summary: CRISPR/Cas9 is a powerful tool for gene editing, but it is challenging to screen modified cells. This study developed two novel traffic light screening reporters, PMG-SSA and PMG-HDR, to measure nuclease activity and select genetically modified cells. The reporters can be self-repaired, resulting in a functional selection cassette that can be used for screening. The results showed improved enrichment efficiencies for gene knockout and knock-in cells. These surrogate reporters advance CRISPR/Cas9-mediated editing in mammalian cells.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2023)
Review
Biochemistry & Molecular Biology
Ali Saber Sichani, Maryam Ranjbar, Maryam Baneshi, Farid Torabi Zadeh, Jafar Fallahi
Summary: In the field of medicine, the development of precise gene-editing tools, particularly CRISPR/Cas9, has revolutionized the potential for gene therapy in treating genetic diseases. This article discusses the different types of CRISPR/Cas-based gene-editing techniques, their functions, and the ongoing research studies that aim to improve their efficiency and reliability for future gene therapies. The article also compares the capabilities of these platforms and highlights their potential limitations.
MOLECULAR BIOTECHNOLOGY
(2023)
Correction
Clinical Neurology
Sebastian Keller, Witold Henryk Polanski, Christoph Enzensperger, Heinz Reichmann, Andreas Hermann, Gabriele Gille
JOURNAL OF NEURAL TRANSMISSION
(2022)
Article
Behavioral Sciences
Anna G. M. Temp, Elisabeth Kasper, Stefan Vielhaber, Judith Machts, Andreas Hermann, Stefan Teipel, Johannes Prudlo
Summary: This study investigated insight into behavioral decline in ALS and ALS-FTD patients, finding that even without FTD, ALS patients experience behavioral decline, while ALS-FTD patients lose awareness of this decline. Unexpectedly, higher IQ was associated with poorer insight into behavioral decline.
BRAIN AND BEHAVIOR
(2022)
Article
Oncology
Julia Suttorp, Jonathan Lukas Luehmann, Yvonne Lisa Behrens, Gudrun Goehring, Doris Steinemann, Dirk Reinhardt, Nils von Neuhoff, Markus Schneider
Summary: Optical Genome Mapping (OGM) is an emerging chip-based DNA technique that can efficiently identify genetic aberrations in pediatric AML. Compared to traditional cytogenetic methods, OGM has higher resolution and shorter turnaround time, and can detect previously unknown alterations. OGM can also serve as a tool for finding new minimal residual disease markers and plays an important role in risk stratification of pediatric AML.
Article
Cell Biology
Rene Guenther, Arun Pal, Chloe Williams, Vitaly L. Zimyanin, Maria Liehr, Clare von Neubeck, Mechthild Krause, Mrudula G. Parab, Susanne Petri, Norman Kalmbach, Stefan L. Marklund, Jared Sterneckert, Peter Munch Andersen, Florian Wegner, Jonathan D. Gilthorpe, Andreas Hermann
Summary: Little is known about the early pathogenic events of mutant SOD1 in ALS. Using a human iPSC model, this study reveals that mitochondrial dysfunction is one of the crucial early steps in the pathogenic cascade of SOD1-ALS.
Article
Cell & Tissue Engineering
Mark-Christian Jaboreck, Jonathan Lukas Luehmann, Mia Mielenz, Frauke Stanke, Gudrun Goehring, Ulrich Martin, Ruth Olmer, Sylvia Merkert
Summary: In this study, knockout cell lines lacking the TMEM16A gene were generated using CRISPR/Cas9 technology in both healthy human induced pluripotent stem cells (iPSCs) and cystic fibrosis patient iPSCs. These cell lines can provide insights into the role of TMEM16A in mucus secretion and production, as well as evaluating its therapeutic potential.
STEM CELL RESEARCH
(2022)
Article
Biochemistry & Molecular Biology
Franziska Alfen, Elena Putscher, Michael Hecker, Uwe Klaus Zettl, Andreas Hermann, Jan Lukas
Summary: Fabry disease is a rare genetic disorder caused by mutations in the GLA gene. This study investigated the impact of these mutations on splicing behavior and found that abnormal splicing can lead to reduced enzyme activity and affect treatment options. Experimental analysis confirmed alternative splicing events for specific mutations, highlighting the importance of splicing phenotype analysis in studying exonic GLA gene mutations.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Genetics & Heredity
Jonathan Lukas Luehmann, Gunnar Schmidt, Bernd Auber, Anke Katharina Bergmann, Oliver Brandau, Anja Louis, Sabine Hentze, Katrin Eisfeld, Brigitte Schlegelberger, Ruediger Klaes, Doris Steinemann
Summary: This study reports a family in which the father has a deletion at 7q11.23 and is affected by Williams-Beuren syndrome (WBS), but the clinically unaffected parents. The genetic testing confirmed a 1.5 Mb deletion on one allele and a reciprocal 1.8 Mb duplication on the other allele in the father, highlighting the importance of genetic counseling in this family.
AMERICAN JOURNAL OF MEDICAL GENETICS PART A
(2023)
Article
Oncology
Juliette Nowak, Marco Bentele, Ivana Kutle, Katharina Zimmermann, Jonathan Lukas Luehmann, Doris Steinemann, Stephan Kloess, Ulrike Koehl, Willi Rossberg, Amed Ahmed, Dirk Schaudien, Lavinia Neubert, Jan-Christopher Kamp, Mark P. Kuehnel, Athanasia Warnecke, Axel Schambach, Michael Morgan
Summary: Despite poor prognosis and survival rates for head and neck squamous cell carcinomas (HNSCC), immunotherapy using CAR-NK cells targeting HER1/EGFR shows promise for HNSCC treatment. However, targeting HER1 alone is not sufficient to eliminate potential cancer stem cells. It is necessary to target multiple tumor-associated antigens to reduce high relapse rates in HNSCC.
Article
Cell Biology
Wonphorn Kandhavivorn, Hannes Glass, Thomas Herrmannsdoerfer, Tobias M. Boeckers, Marc Uhlarz, Jonas Gronemann, Richard H. W. Funk, Jens Pietzsch, Arun Pal, Andreas Hermann
Summary: Amyotrophic lateral sclerosis (ALS) is a devastating motoneuron disease characterized by loss of neuromuscular junctions, degeneration of motoneurons and muscle paralysis. Magnetic stimulation has been found to significantly improve axonal trafficking and regenerative sprouting of motoneurons in vitro, indicating the therapeutic potential of magnetic stimulations in ALS. However, further long-term in vivo studies are needed for validation.
Article
Cell Biology
Vitaly L. Zimyanin, Anna-Maria Pielka, Hannes Glass, Julia Japtok, Dajana Grossmann, Melanie Martin, Andreas Deussen, Barbara Szewczyk, Chris Deppmann, Eli Zunder, Peter M. Andersen, Tobias M. Boeckers, Jared Sterneckert, Stefanie Redemann, Alexander Storch, Andreas Hermann
Article
Cell Biology
Bita Massih, Alexander Veh, Maren Schenke, Simon Mungwa, Bettina Seeger, Bhuvaneish T. Selvaraj, Siddharthan Chandran, Peter Reinhardt, Jared Sterneckert, Andreas Hermann, Michael Sendtner, Patrick Lueningschroer
Summary: This article introduces a human neuromuscular co-culture system that enables the formation of neuromuscular junctions by differentiating induced pluripotent stem cells into motor neurons and 3D skeletal muscle tissue. The system is suitable for modeling the pathophysiology of motor neuron diseases and has been used to study the synaptic coupling and muscle contraction in Amyotrophic Lateral Sclerosis (ALS). This in vitro model recapitulates aspects of human physiology and is of great importance for studying human motor neurons in health and disease.
FRONTIERS IN CELL AND DEVELOPMENTAL BIOLOGY
(2023)
Letter
Oncology
Zeljko Antic, Alena van Boemmel, Konstantin Riege, Jana Lentes, Charlotte Schroeder, Julia Alten, Cornelia Eckert, Lara Fuhrmann, Doris Steinemann, Lennart Lenk, Denis M. Schewe, Martin Zimmermann, Martin Schrappe, Brigitte Schlegelberger, Gunnar Cario, Steve Hoffmann, Anke K. Bergmann
Editorial Material
Medicine, General & Internal
Matthias Boentert, Andreas Hermann, Julian Grosskreutz
JOURNAL OF CLINICAL MEDICINE
(2023)
Article
Cell Biology
Cristina Marisol Castillo Bautista, Kristin Eismann, Marc Gentzel, Silvia Pelucchi, Jerome Mertens, Hannah E. Walters, Maximina H. Yun, Jared Sterneckert
Summary: Aging disrupts protein homeostasis and contributes to various diseases, including ALS. Repressing autophagy is a strategy to restore protein homeostasis and protect neurons. A study found that obatoclax, a small molecule BH3 mimetic, can disrupt the interaction between BECN1 and BCL2, reducing FUS levels and improving protein homeostasis to rescue neurons. Obatoclax has potential as a therapeutic for ALS and other age-related disorders linked to protein homeostasis defects.
Article
Cell Biology
Francesco Antoniani, Marco Cimino, Laura Mediani, Jonathan Vinet, Enza M. Verde, Valentina Secco, Alfred Yamoah, Priyanka Tripathi, Eleonora Aronica, Maria E. Cicardi, Davide Trotti, Jared Sterneckert, Anand Goswami, Serena Carra
Summary: Amyotrophic Lateral Sclerosis (ALS) and Frontotemporal Dementia (FTD) are neurodegenerative disorders linked by genetic causes, with TARDBP, FUS, and C9orf72 genes playing critical roles. The proteins produced by these genes accumulate in the cytoplasm, leading to pathological inclusions. This study explores the connection between the cytoplasmic and nuclear protein quality control (PQC) systems, highlighting the impairment of PML nuclear bodies (PML-NBs) and cytoplasmic stress granules (SGs) in ALS-FTD cases, which contribute to protein aggregation.
CELL DEATH DISCOVERY
(2023)