Article
Biochemistry & Molecular Biology
Samantha Baldassarri, Daniela Benati, Federica D'Alessio, Clarissa Patrizi, Eleonora Cattin, Michela Gentile, Angelo Raggioli, Alessandra Recchia
Summary: This study demonstrates the use of SB transposon to engineer HEK293 cells for improved production of chimpanzee Adenovector. By employing genetic engineering and CRISPR/Cas9 technology, the essential genes for chimpanzee Adenoviral DNA replication were successfully integrated into HEK293 cells, leading to significantly enhanced vector production.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Review
Biochemistry & Molecular Biology
Nicolas Sandoval-Villegas, Wasifa Nurieva, Maximilian Amberger, Zoltan Ivics
Summary: Transposons are mobile genetic elements that have been used as experimental tools to introduce foreign DNA sequences into target cells in a regulated and highly efficient manner. They are supplied as naked nucleic acids or recombinant proteins, making their use simple, safe, and economically competitive. Transposons enable various avenues for genome manipulations in vertebrates.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2021)
Article
Biochemistry & Molecular Biology
Natalie Tschorn, Yasemin van Heuvel, Jorn Stitz
Summary: The use of two-component transposon plasmid vector systems can expedite the development of recombinant cell lines, but the undesired stable transfection and sustained expression of the transposase construct can lead to genetic instability. In this study, we established three recombinant cell pools using a Sleeping Beauty-derived vector system and demonstrated stable integration and sustained expression of the transposase for 48 days. To provide an alternative approach, we generated transcripts of the transposase gene in vitro and co-transfected them with donor vector plasmids, resulting in high copy number integrations and expression levels of the gene of interest (GOI). We anticipate that the use of transposase mRNA will contribute to further improvements in cell line development processes.
MOLECULAR BIOTECHNOLOGY
(2023)
Article
Biotechnology & Applied Microbiology
Yun Haeng Lee, Ji Yun Park, Eun Seon Song, Haneur Lee, Myeong Uk Kuk, Junghyun Joo, Hyungmin Roh, Joon Tae Park
Summary: This study proposes a new strategy to increase the efficiency and stability of protein production by modifying the traditional SB transposon vector. Adding a pair of inverted terminal repeats (ITRs) next to existing ITRs significantly improves transgene integration efficiency, resulting in high-yield and sustained protein production. Moreover, double-ITRs show a more favorable response to DNA methylation inhibitors, suggesting their potential in increasing protein productivity.
BIOTECHNOLOGY AND BIOPROCESS ENGINEERING
(2022)
Review
Hematology
Fernando Anjos-Afonso, Dominique Bonnet
Summary: The ability to isolate and characterize different hematopoietic stem cell (HSC) or progenitor cell populations provides insights into the regulation of hematopoiesis in various conditions. While significant progress has been made in understanding the composition of these cell types in mice, recent breakthroughs have improved the resolution of the human primitive hematopoietic compartment. This review aims to provide a historical perspective and discuss the progress in characterizing human postnatal CD34(+) HSC-enriched populations, with potential implications for future translational studies.
Article
Genetics & Heredity
Sarah C. Saunderson, S. M. Ali Hosseini-Rad, Alexander D. McLellan
Summary: Tetracycline-inducible systems are widely used control elements for mammalian gene expression. The G72V mutation in the rtTA-M2 tetracycline activator reduces basal level expression and increases fold-induction. Sensitivity enhancing mutations compensate for the loss of responsiveness caused by the G72V mutation without compromising promoter tightness.
Article
Biochemistry & Molecular Biology
Aiping Zhang, Lijian Wang, Josh Haipeng Lei, Zhengqiang Miao, Monica Vishnu Valecha, Peng Hu, Kai Miao, Chu-Xia Deng
Summary: SB Digestor is a new computational algorithm that overcomes biological heterogeneity and accurately detects and defines cancer driver genes, providing more potential candidates for study.
INTERNATIONAL JOURNAL OF BIOLOGICAL SCIENCES
(2023)
Review
Oncology
Haruna Takeda, Nancy A. Jenkins, Neal G. Copeland
Summary: Cancer genome sequencing studies have identified driver genes for various cancers, but identifying these genes solely from genetic data remains challenging. In vivo forward genetic screens using SB transposon mutagenesis in mice provide a powerful tool for identifying potential cancer driver genes. Comparing driver genes identified in human and mouse tumors can enhance confidence in identifying cancer driver genes.
Article
Genetics & Heredity
Yiting Zhou, Guangwei Ma, Jiawen Yang, Zenghong Gao, Yabin Guo
Summary: Recent study showed that Sleeping Beauty transposon can integrate into non-TA sites, mainly through aberrant integrations. The consensus sequence of the non-TA target sites corresponds to the transposon end sequence.
FRONTIERS IN GENETICS
(2021)
Review
Virology
Matthias T. Ochmann, Zoltan Ivics
Summary: The Sleeping Beauty transposon system has been widely used as a genetic engineering tool, with extensive studies elucidating the molecular mechanisms and actions during SB transposition process. Structural data on the SB transposase provide a direct insight into the enzyme's functionality, leading to the development of novel SB variants for advanced genetic engineering possibilities. However, many aspects of transposition process remain poorly understood, necessitating further research to explore the structure-function relationships of SB transposition for the development of new vector systems.
Article
Medicine, Research & Experimental
Kun Ma, Wenzhe Li, Guang Zhu, Shibo Sun, Hao Chi, Yalin Yin, He Diao, Xiao-Jin Xing, Zhaoming Guo, Li Wang, Weiping Xu, Changhao Cui, Jianqiang Xu
Summary: The CRISPR/Cas9 gene was cloned into a non-viral safe vector Sleeping-Beauty (SB) transposon to construct a safer vector. In addition, PDA/DEX-PEI@HA (PDPH) nanoparticles were developed to facilitate precise CRISPR/Cas9 targeted delivery, showing high transfection efficiency and minimal cytotoxicity. The study highlights the potential of PDA/DEX-PEI25k@HA/pT25pCas9 (PDPH25k/pT(25K)pCas9) as a non-viral gene vector for CRISPR/Cas9 delivery in clinical medication.
BIOMEDICINE & PHARMACOTHERAPY
(2021)
Article
Cell Biology
Lea Flippe, Anne Gaignerie, Celine Serazin, Olivier Baron, Xavier Saulquin, Ignacio Anegon, Laurent David, Carole Guillonneau
Summary: Immunotherapy using primary T cells has greatly improved medical care in recent years, but limitations such as challenging cell genome editing and lack of standardized products have restricted its clinical use. The use of T cells derived from human pluripotent stem cells offers advantages of genetic modification and standardized allogeneic products. However, more research is needed to fully understand the feasibility and functionality of these T cells before clinical applications. This study successfully generated T cells from induced pluripotent stem cells, demonstrating the potential for therapeutic use and treatment of various diseases.
Article
Multidisciplinary Sciences
Eliot Y. Zhu, Jacob L. Schillo, Sarina D. Murray, Jesse D. Riordan, Adam J. Dupuy
Summary: Combined BRAF and MEK inhibition is an effective treatment for BRAF-mutant cutaneous melanoma, but drug resistance remains a challenge. This study utilized the Sleeping Beauty transposon system to investigate the mechanisms of drug resistance in melanoma. The findings demonstrated the importance of considering melanoma heterogeneity and identified a potential target for overcoming drug resistance.
Article
Cell & Tissue Engineering
Paul Jaeger, Stefanie Geyh, Soeren Twarock, Ron-Patrick Cadeddu, Pablo Rabes, Annemarie Koch, Uwe Maus, Tobias Hesper, Christoph Zilkens, Christina Rautenberg, Felix Bormann, Karl Koehrer, Patrick Petzsch, Dagmar Wieczorek, Beate Betz, Harald Surowy, Barbara Hildebrandt, Ulrich Germing, Guido Kobbe, Rainer Haas, Thomas Schroeder
Summary: Studies suggest that leukemic cells induce functional inhibition of healthy CD34+ HSPC, at least in part, through pathways like TGF beta 1, indicating that blocking this pathway may improve hematopoiesis in AML patients.
Article
Immunology
Sabrina B. Bennstein, Sandra Weinhold, Oezer Degistirici, Robert A. J. Oostendorp, Katharina Raba, Gesine Koegler, Roland Meisel, Lutz Walter, Markus Uhrberg
Summary: ILC3, particularly IL-22-secreting ILC3, play vital roles in maintaining mucosal barrier functions and controlling inflammatory diseases. A method for generating functionally competent human ILC3 from cord blood-derived CD34(+) hematopoietic progenitors on human MSCs has been developed, showing promise for potential clinical applications in treating inflammatory diseases and cancer.
FRONTIERS IN IMMUNOLOGY
(2021)
Editorial Material
Hematology
Donald B. Kohn
Article
Cell & Tissue Engineering
Jason P. Awe, Eric H. Gschweng, Agustin Vega-Crespo, Jon Voutila, Mary H. Williamson, Brian Truong, Donald B. Kohn, Noriyuki Kasahara, James A. Byrne
STEM CELLS TRANSLATIONAL MEDICINE
(2015)
Review
Hematology
Donald B. Kohn, Matthew H. Porteus, Andrew M. Scharenberg
Review
Multidisciplinary Sciences
Cynthia E. Dunbar, Katherine A. High, J. Keith Joung, Donald B. Kohn, Keiya Ozawa, Michel Sadelain
Article
Biochemistry & Molecular Biology
Peirong Hui, Yanmin Bi, Hong Ma, Thipparat Suwanmanee, Brian Zeithaml, Nate J. Fry, Donald B. Kohn, Tal Kafri
Article
Biochemistry & Molecular Biology
Aaron R. Cooper, Georgia R. Lill, Eric H. Gschweng, Donald B. Kohn
NUCLEIC ACIDS RESEARCH
(2015)
Article
Cell & Tissue Engineering
Sandeep Soni, Donald B. Kohn
Review
Biochemical Research Methods
Donald B. Kohn
CURRENT OPINION IN BIOTECHNOLOGY
(2019)
Article
Biochemistry & Molecular Biology
Donald B. Kohn, Claire Booth, Elizabeth M. Kang, Sung-Yun Pai, Kit L. Shaw, Giorgia Santilli, Myriam Armant, Karen F. Buckland, Uimook Choi, Suk See De Ravin, Morna J. Dorsey, Caroline Y. Kuo, Diego Leon-Rico, Christine Rivat, Natalia Izotova, Kimberly Gilmour, Katie Snell, Jinhua Xu-Bayford Dip, Jinan Darwish, Emma C. Morris, Dayna Terrazas, Leo D. Wang, Christopher A. Bauser, Tobias Paprotka, Douglas B. Kuhns, John Gregg, Hayley E. Raymond, John K. Everett, Geraldine Honnet, Luca Biasco, Peter E. Newburger, Frederic D. Bushman, Manuel Grez, H. Bobby Gaspar, David A. Williams, Harry L. Malech, Anne Galy, Adrian J. Thrasher, Karen F. Buckland, Christopher A. Bauser, Geraldine Honnet, Manuel Grez, H. Bobby Gaspar, Anne Galy, Adrian J. Thrasher
Article
Genetics & Heredity
David H. Gray, Isaac Villegas, Joseph Long, Jasmine Santos, Alexandra Keir, Alison Abele, Caroline Y. Kuo, Donald B. Kohn
Summary: X-linked agammaglobulinemia (XLA) is a genetic disorder characterized by low immunoglobulin levels and increased infection risks. Gene editing in hematopoietic stem cells of XLA patients aims to correct the Bruton's tyrosine kinase (BTK) gene mutation and restore B cell development. The CRISPR-Cas9 platform enables precise integration of corrective BTK cDNA, potentially offering therapeutic levels of gene therapy for XLA.
Article
Immunology
Carolyn H. Baloh, Samiksha A. Borkar, Kai-Fen Chang, Jiqiang Yao, Michael S. Hershfield, Suhag H. Parikh, Donald B. Kohn, Maureen M. Goodenow, John W. Sleasman, Li Yin
Summary: Based on deep sequencing, gene therapy resulted in an IgHV repertoire with molecular diversity similar to healthy infants.
JOURNAL OF CLINICAL IMMUNOLOGY
(2021)
Article
Oncology
Jiaying Han, Kevin Tam, Curtis Tam, Roger P. Hollis, Donald B. Kohn
Summary: Lentiviral vectors are efficient tools for gene therapy, but their production can be hindered by limitations such as low titers and reduced gene transfer capacity. This study identified host restriction factors in packaging cells and developed a CRISPRed HEK293T cell line to disrupt these factors, leading to significant increases in viral titers. Overexpression of transcription elongation factors SPT4 and SPT5 during packaging also improved LV production. These approaches have the potential to benefit various gene therapy applications.
MOLECULAR THERAPY-ONCOLYTICS
(2021)
Article
Hematology
Shanna L. White, Thomas D. Lee, Traci Toy, Judith E. Carroll, Lilian Polsky, Beatriz Campo Fernandez, Alejandra Davila, Donald B. Kohn, Vivian Y. Chang
Summary: The study investigated the impact of autologous stem cell transplant with gene therapy on telomere lengths (TL) and clonal hematopoiesis of indeterminate potential (CHIP) in pediatric patients with adenosine deaminase-deficient severe combined immune deficiency. The findings suggest the need for further research to reveal the underlying genetic risk of malignancy in participants undergoing gene therapy.
Review
Immunology
Donald B. Kohn
Summary: ADA SCID is a severe inborn immune deficiency disease that can be effectively treated using gene therapy. Continuous improvements in treatment methods have led to significant immune restoration and stable engraftment of gene-corrected hematopoietic stem cells in most ADA SCID patients. However, achieving sustained availability and access to this therapy remains challenging.
IMMUNOLOGICAL REVIEWS
(2023)
Meeting Abstract
Immunology
Lili Yang, Drake J. Smith, Levina J. Lin, Heesung Moon, Alexander T. Pham, Xi Wang, Siyuan Liu, Sunjong Ji, Valerie Rezek, Saki Shimizu, Marlene Ruiz, Jennifer Lam, Deanna M. Janzen, Sanaz Memarzadeh, Donald B. Kohn, Jerome A. Zack, Scott G. Kitchen, Dong Sung An
JOURNAL OF IMMUNOLOGY
(2017)