646 results match your criteria piggybac transposon


Development of non-viral, ligand-dependent, EPHB4-specific chimeric antigen receptor T cells for treatment of rhabdomyosarcoma.

Mol Ther Oncolytics 2021 Mar 5;20:646-658. Epub 2021 Mar 5.

Department of Pediatrics, Graduate School of Medicine, Kyoto Prefectural University of Medicine, Kyoto 602-8566, Japan.

Ephrin type-B receptor 4 (EPHB4), expressed in tumors including rhabdomyosarcoma, is a suitable target for chimeric antigen receptor (CAR)-T cells. Ligand-independent activation of EPHB4 causes cell proliferation and malignant transformation in rhabdomyosarcoma, whereas ligand-dependent stimulation of EPHB4 induces apoptosis in rhabdomyosarcoma. Therefore, we hypothesized that ligand-based, EPHB4-specific CAR-T cells may kill rhabdomyosarcoma cells without stimulating downstream cell proliferation mechanisms. Read More

View Article and Full-Text PDF

Manufacturing NKG2D CAR-T cells with piggyBac transposon vectors and K562 artificial antigen-presenting cells.

Mol Ther Methods Clin Dev 2021 Jun 3;21:107-120. Epub 2021 Mar 3.

Department of Biological Sciences, National University of Singapore, Singapore 117543, Singapore.

Non-viral platforms can be applied rapidly and cost-effectively for chimeric antigen receptor (CAR)-T cell manufacturing. In the present paper, we describe in detail a clinically relevant manufacturing process for NKG2D CAR-T cells through electroporation of CAR-encoding piggyBac transposon plasmids and expansion with K562 artificial antigen-presenting cells. With an optimized protocol, we generated the final cell therapy products with 89. Read More

View Article and Full-Text PDF

Generation of WAe001-A-58 human embryonic stem cell line with inducible expression of the SARS-CoV-2 nucleocapsid protein.

Stem Cell Res 2021 Jan 26:102197. Epub 2021 Jan 26.

Institute of Public Health, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China; Guangdong Provincial Key Laboratory of Biocomputing, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China; Key Laboratory of Regenerative Biology, South China Institute for Stem Cell Biology and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, Guangzhou, China. Electronic address:

Excessive prostaglandin E (PGE) is the key pathological basis for COVID-19 and a Celebrex treatment of hospitalized COVID-19 patients with comorbidities led to 100% discharged rate and zero death (Hong et al. 2020). It is also suggested that SARS-CoV-2 infected multiple organs and the SARS-CoV nucleocapsid (N) protein transcriptionally drives the expression of the host COX-2 gene. Read More

View Article and Full-Text PDF
January 2021

Generation of Murine Induced Pluripotent Stem Cells through Transposon-Mediated Reprogramming.

Methods Mol Biol 2021 Mar 24. Epub 2021 Mar 24.

Department of Biotechnology, Friedrich-Loeffler-Institut, Federal Research Institute for Animal Health, Neustadt, Germany.

The seminal discovery of induced pluripotent stem (iPS) cells through ectopic expression of a cocktail of gene factors (OCT4, SOX2, KLF4, and c-MYC) by the group of Yamanaka was a major breakthrough, gained widespread acclaim and garnered much attention in the field of stem cell science. The iPS cells possess most of the characteristics and advantages of embryonic stem (ES) cells without the association of ethical stigma for their derivation. In addition, these cells can give rise to any cell type of the body and thus have tremendous potential for many downstream applications in research and regenerative medicine. Read More

View Article and Full-Text PDF

The NFIB-ERO1A axis promotes breast cancer metastatic colonization of disseminated tumour cells.

EMBO Mol Med 2021 Apr 10;13(4):e13162. Epub 2021 Mar 10.

Department of Biomedicine, Department of Surgery, University Hospital Basel, University of Basel, Basel, Switzerland.

Metastasis is the main cause of deaths related to solid cancers. Active transcriptional programmes are known to regulate the metastatic cascade but the molecular determinants of metastatic colonization remain elusive. Using an inducible piggyBac (PB) transposon mutagenesis screen, we have shown that overexpression of the transcription factor nuclear factor IB (NFIB) alone is sufficient to enhance primary mammary tumour growth and lung metastatic colonization. Read More

View Article and Full-Text PDF

JMJD6 negatively regulates cytosolic RNA induced antiviral signaling by recruiting RNF5 to promote activated IRF3 K48 ubiquitination.

PLoS Pathog 2021 Mar 8;17(3):e1009366. Epub 2021 Mar 8.

State Key Laboratory of Veterinary Etiological Biology, National Foot and Mouth Diseases Reference Laboratory, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, China.

The negative regulation of antiviral immune responses is essential for the host to maintain homeostasis. Jumonji domain-containing protein 6 (JMJD6) was previously identified with a number of functions during RNA virus infection. Upon viral RNA recognition, retinoic acid-inducible gene-I-like receptors (RLRs) physically interact with the mitochondrial antiviral signaling protein (MAVS) and activate TANK-binding kinase 1 (TBK1) to induce type-I interferon (IFN-I) production. Read More

View Article and Full-Text PDF

Overexpression of Long Non-coding RNA 4933425B07 Causes Urinary Malformations in Mice.

Front Cell Dev Biol 2021 19;9:594640. Epub 2021 Feb 19.

Department of Nephrology, Shanghai Kidney Development and Pediatric Kidney Disease Research Center, Children's Hospital of Fudan University, Shanghai, China.

Congenital anomalies of the kidney and urinary tract (CAKUT) is a common birth defect and is the leading cause of end-stage renal disease in children. The etiology of CAKUT is complex and includes mainly genetic and environmental factors. However, these factors cannot fully explain the etiological mechanism of CAKUT. Read More

View Article and Full-Text PDF
February 2021

Human epidermal growth factor-functionalized cocoon silk with improved cell proliferation activity for the fabrication of wound dressings.

J Biomater Appl 2021 Mar 4:885328221997981. Epub 2021 Mar 4.

College of Animal Science, 12377Zhejiang University, Hangzhou, Zhejiang, China.

Human epidermal growth factor (hEGF) is a key factor involved in wound healing owing to its powerful ability to stimulate cell proliferation. In this study, we used piggyBac transposon technology to produce transgenic silkworms expressing the hEGF protein fused to truncated heavy chain (FibH-hEGF). The FibH-hEGF fusion protein was successfully expressed and secreted into silkworm cocoons. Read More

View Article and Full-Text PDF

[Preparation of transgenic Musca domestica by microinjection method].

Sheng Wu Gong Cheng Xue Bao 2021 Feb;37(2):655-662

Key Laboratory of Biotechnology and Medical Engineering, School of Biology and Engineering, Guizhou Medical University, Guiyang 550025, Guizhou, China.

The transposon vector containing enhanced green fluorescent protein (EGFP) was injected into early housefly (Musca domestica L.) eggs by microinjection method to realize stable gene expression in vivo for verification, and to study housefly gene function. A borosilicate glass micro injection needle suitable for microinjection of housefly eggs was made, the softening treatment conditions of housefly egg shells were explored, and a microinjection technology platform suitable for housefly was constructed with a high-precision microsyringe Nanoject Ⅲ as the main body. Read More

View Article and Full-Text PDF
February 2021

A native, highly active Tc1/mariner transposon from zebrafish (ZB) offers an efficient genetic manipulation tool for vertebrates.

Nucleic Acids Res 2021 02;49(4):2126-2140

College of Animal Science & Technology, Yangzhou University, Yangzhou, Jiangsu 225009, China.

New genetic tools and strategies are currently under development to facilitate functional genomics analyses. Here, we describe an active member of the Tc1/mariner transposon superfamily, named ZB, which invaded the zebrafish genome very recently. ZB exhibits high activity in vertebrate cells, in the range of those of the widely used transposons piggyBac (PB), Sleeping Beauty (SB) and Tol2. Read More

View Article and Full-Text PDF
February 2021

Chimeric antigen receptor T cells engineered to secrete CD40 agonist antibodies enhance antitumor efficacy.

J Transl Med 2021 02 18;19(1):82. Epub 2021 Feb 18.

Department of Biotherapy, Eastern Hepatobiliary Surgery Hospital, Navy Medical University, Shanghai, 201805, China.

Background: Although chimeric antigen receptor (CAR)-T cell therapy has been remarkably successful for haematological malignancies, its efficacy against solid tumors is limited. The combination of CAR-T cell therapy with immune checkpoint inhibitors (CPIs), such as PD-1, PD-L1, and CTLA-4 antibodies, is a promising strategy for enhancing the antitumor efficacy of CAR-T cells. However, because most patients acquire resistance to CPIs, investigating other strategies is necessary to further improve the antitumor efficacy of CAR-T cell therapy for solid tumors. Read More

View Article and Full-Text PDF
February 2021

A piggyBac-mediated transgenesis system for the temporary expression of CRISPR/Cas9 in rice.

Plant Biotechnol J 2021 Feb 2. Epub 2021 Feb 2.

Plant Genome Engineering Research Unit, Institute of Agrobiological Sciences, National Agriculture and Food research Organization (NARO), Tsukuba, Japan.

Targeted mutagenesis via CRISPR/Cas9 is now widely used, not only in model plants but also in agriculturally important crops. However, in vegetative crop propagation, CRISPR/Cas9 expression cassettes cannot be segregated out in the resulting progenies, but must nevertheless be eliminated without leaving unnecessary sequences in the genome. To this end, we designed a piggyBac-mediated transgenesis system for the temporary expression of CRISPR/Cas9 in plants. Read More

View Article and Full-Text PDF
February 2021

Stabilization of Poly (β-Amino Ester) Nanoparticles for the Efficient Intracellular Delivery of PiggyBac Transposon.

Bioengineering (Basel) 2021 Jan 20;8(2). Epub 2021 Jan 20.

Department of Biomedical Engineering and Chemical Engineering, University of Texas at San Antonio, One UTSA Circle, San Antonio, TX 78249, USA.

The administration of gene-editing tools has been proposed as a promising therapeutic approach for correcting mutations that cause diseases. Gene-editing tools, composed of relatively large plasmid DNA constructs that often need to be co-delivered with a guiding protein, are unable to spontaneously penetrate mammalian cells. Although viral vectors facilitate DNA delivery, they are restricted by the size of the plasmid to carry. Read More

View Article and Full-Text PDF
January 2021

The C-terminal Domain of piggyBac Transposase Is Not Required for DNA Transposition.

J Mol Biol 2021 Apr 13;433(7):166805. Epub 2021 Jan 13.

PRC, UMR INRAE 0085, CNRS 7247, Centre INRAE Val de Loire, 37380 Nouzilly, France. Electronic address:

PiggyBac(PB)-like elements (pble) are members of a eukaryotic DNA transposon family. This family is of interest to evolutionary genomics because pble transposases have been domesticated at least 9 times in vertebrates. The amino acid sequence of pble transposases can be split into three regions: an acidic N-terminal domain (~100 aa), a central domain (~400 aa) containing a DD[D/E] catalytic triad, and a cysteine-rich domain (CRD; ~90 aa). Read More

View Article and Full-Text PDF

Generation of a human embryonic stem cell (WAe001-A-47) with hVGLL4 doxycyclin-inducible expression by the PiggyBac transposon system.

Stem Cell Res 2020 Dec 24;50:102142. Epub 2020 Dec 24.

Institute of Hypoxia Medicine, School of Basic Medical Sciences, Wenzhou Medical University, Wenzhou 325035, Zhejiang, China. Electronic address:

VGLL4 is a new component of the Hippo pathway and bind TEADs to compete with YAP, so as to inhibit tumor progression, but its role in stem cell and organ regeneration remains unclear. Using the PiggyBac transposon system, we generated a VGLL4 doxycycline-inducible expression human embryonic stem cell line (WAe001-A-47). The established hESC line retains its normal morphology and pluripotency markers with in vitro differentiation potential, as well as a normal karyotype. Read More

View Article and Full-Text PDF
December 2020

Improvement on the genetic engineering of an invasive agricultural pest insect, the cherry vinegar fly, Drosophila suzukii.

BMC Genet 2020 12 18;21(Suppl 2):139. Epub 2020 Dec 18.

Department of Developmental Biology, Johann-Friedrich-Blumenbach-Institute of Zoology and Anthropology, Göttingen Center for Molecular Biosciences, Georg-August-University Göttingen, 37077, Göttingen, Germany.

Background: The invasive fly Drosophila suzukii has become an established fruit pest in Europe, the USA, and South America with no effective and safe pest management. Genetic engineering enables the development of transgene-based novel genetic control strategies against insect pests and disease vectors. This, however, requires the establishment of reliable germline transformation techniques. Read More

View Article and Full-Text PDF
December 2020

Is Use of BMP-2 Associated with Tumor Growth and Osteoblastic Differentiation in Murine Models of Osteosarcoma?

Clin Orthop Relat Res 2020 12;478(12):2921-2933

J. K. Kendal, M. J. Monument, Section of Orthopaedic Surgery, Department of Surgery, University of Calgary, Calgary, Canada.

Background: The putative benefit of rhBMP-2 is in the setting of limb reconstruction using structural allografts, whether it be allograft-prosthetic composites, osteoarticular allografts, or intercalary segmental grafts. There are also potential advantages in augmenting osseointegration of uncemented endoprosthetics and in reducing infection. Recombinant human BMP-2 might mitigate nonunion in structural allograft augmented osteosarcoma limb salvage surgery; however, its use is limited because of concerns about the prooncogenic effects of the agent. Read More

View Article and Full-Text PDF
December 2020

Autologous non-human primate model for safety assessment of transposon-mediated chimeric antigen receptor T cells on granulocyte-macrophage colony-stimulating factor receptor.

Clin Transl Immunology 2020 22;9(11):e1207. Epub 2020 Nov 22.

Department of Pediatrics Shinshu University School of Medicine Matsumoto Japan.

Objectives: Chimeric antigen receptor (CAR)-T cell therapy redirected to specific antigens on tumor cells is a promising immunotherapy strategy for various cancers. Most target antigens are also expressed on normal tissues at varying levels, and therefore, a considerable challenge in the field is determining safety profiles, including life-threatening off-tumor and off-target toxicities. The granulocyte-macrophage colony-stimulating factor receptor (hGMR) is a promising target for CAR T-cell therapy for a subset of acute myelocytic leukaemia, although it is also expressed on normal cells including monocytes, macrophages, CD34-positive haematopoietic cells and vascular endothelial cells. Read More

View Article and Full-Text PDF
November 2020

Transcriptome sequencing reveals Cnaphalocrocis medinalis against baculovirus infection by oxidative stress.

Mol Immunol 2021 01 20;129:63-69. Epub 2020 Nov 20.

Jiangsu Lixiahe District Institute of Agricultural Sciences, Yangzhou, 225007, China. Electronic address:

Cnaphalocrocis medinalis granulovirus (CnmeGV) is a potential microbial agent against the rice leaffolder. Innate immunity is essential for insects to survive pathogenic infection. Therefore, to clarify the immune response of Cnaphalocrocis medinalis to the viral colonization, the gene expression profile of C. Read More

View Article and Full-Text PDF
January 2021

Evaluation of piggyBac-mediated anti-CD19 CAR-T cells after ex vivo expansion with aAPCs or magnetic beads.

J Cell Mol Med 2021 Jan 22;25(2):686-700. Epub 2020 Nov 22.

Central Laboratory of Yan'an Hospital Affiliated to Kunming Medical University, Kunming, China.

Adoptive immunotherapy is a new potential method of tumour therapy, among which anti-CD19 chimeric antigen receptor T-cell therapy (CAR-T cell), is a typical treatment agent for haematological malignancies. Previous clinical trials showed that the quality and phenotype of CAR-T cells expanded ex vivo would seriously affect the tumour treatment efficacy. Although magnetic beads are currently widely used to expand CAR-T cells, the optimal expansion steps and methods have not been completely established. Read More

View Article and Full-Text PDF
January 2021

Unpredictable recombination of PB transposon in Silkworm: a potential risk.

Mol Genet Genomics 2021 Mar 17;296(2):271-277. Epub 2020 Nov 17.

State Key Laboratory of Silkworm Genome Biology, College of Textile and Garment, Southwest University, No. 2 Tiansheng Street, Beibei, Chongqing, 400715, China.

The piggyBac (PB) transposon is the most widely used vector for generating transgenic silkworms. The stability of the PB transposon in the receptor is a serious concern that requires attention because of biosafety concerns. In this study, we found that the transgene silkworm developed loss of reporter gene traits. Read More

View Article and Full-Text PDF

Characterization of bovine interleukin-2 stably expressed in HEK-293 cells.

J Vet Med Sci 2021 Jan 11;83(1):134-141. Epub 2020 Nov 11.

Department of Veterinary Sciences, Faculty of Agriculture, University of Miyazaki, 1-1 Gakuen-Kibanadai-Nishi, Miyazaki 889-2192, Japan.

Interleukin 2 (IL-2) is a pleotropic cytokine and well-known as a T cell growth factor in immunology. It is now known to exert both immunostimulatory and immunosuppressive effects, optimizing immunological microenvironments for effector and regulatory T cell responses. The immunomodulatory role of IL-2 is critical for deciding whether or not T cell responses against specific antigens result in protection. Read More

View Article and Full-Text PDF
January 2021

Novel application of adipose-derived mesenchymal stem cells via producing antiangiogenic factor TSP-1 in lung metastatic melanoma animal model.

Biologicals 2020 Nov 5;68:9-18. Epub 2020 Oct 5.

Departments of Applied Cell Sciences, Faculty of Medicine, Kashan University of Medical Sciences, Kashan, Iran.

Human adipose tissue derived mesenchymal stem cells (hAD-MSC) with suppressive immunogenicity, homing to injury, inflammatory, and cancer sites can be suitable for gene therapy. PiggyBac (PB) is a type of transposon vector applied in mammalian systems and could overcome some limitations of other transposon and viral vectors. In this study, the therapeutic potential hAD-MSCs expressing thrombospondin-1 (TSP-1) is assessed through tail vein injection in C57BL/6 models bearing melanoma mice. Read More

View Article and Full-Text PDF
November 2020

Use of a Hybrid Adeno-Associated Viral Vector Transposon System to Deliver the Insulin Gene to Diabetic NOD Mice.

Cells 2020 10 2;9(10). Epub 2020 Oct 2.

School of Life Sciences, University of Technology Sydney, 15 Broadway, Ultimo, NSW 2007, Australia.

Previously, we used a lentiviral vector to deliver furin-cleavable human insulin () to the livers in several animal models of diabetes using intervallic infusion in full flow occlusion (FFO), with resultant reversal of diabetes, restoration of glucose tolerance and pancreatic transdifferentiation (PT), due to the expression of beta (β)-cell transcription factors (β-TFs). The present study aimed to determine whether we could similarly reverse diabetes in the non-obese diabetic (NOD) mouse using an adeno-associated viral vector (AAV) to deliver - ± the β-TF to the livers of diabetic mice. The traditional AAV8, which provides episomal expression, and the hybrid AAV8/ that results in transgene integration were used. Read More

View Article and Full-Text PDF
October 2020

Precision genome editing in plants via gene targeting and subsequent break-induced single-strand annealing.

Plant Biotechnol J 2021 Mar 26;19(3):563-574. Epub 2020 Oct 26.

Plant Genome Engineering Research Unit, Institute of Agrobiological Sciences, National Agriculture and Food Research Organization, Tsukuba, Japan.

Genome editing via artificial nucleases such as CRISPR/Cas9 has become popular in plants now. However, small insertions or deletions are major mutations and nucleotide substitutions rarely occur when DNA cleavage is induced. To induce nucleotide substitutions, a base editor utilizing dead or nickase-type Cas9 fused with deaminase have been developed. Read More

View Article and Full-Text PDF

Safety and Efficacy of an Immune Cell-Specific Chimeric Promoter in Regulating Anti-PD-1 Antibody Expression in CAR T Cells.

Mol Ther Methods Clin Dev 2020 Dec 14;19:14-23. Epub 2020 Aug 14.

College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou 310018, China.

T cells modified to co-express cytokine or other factors with chimeric antigen receptor (CAR) can induce substantial and persistent increases in antitumor capacity . However, the uncontrolled expression of cytokines or factors can lead to the overactivation of immune cells, causing severe adverse events such as cytokine release syndrome (CRS) and neurotoxicity by CAR T cells with excessive growth potential. Conventional promoters are unregulated, and their expression is unlimited in T cells. Read More

View Article and Full-Text PDF
December 2020

Inducible secretion of IL-21 augments anti-tumor activity of piggyBac-manufactured chimeric antigen receptor T cells.

Cytotherapy 2020 12 17;22(12):744-754. Epub 2020 Sep 17.

Institute of Hematology and Blood Transfusion, Prague, Czech Republic; First Department of Medicine, Department of Haematology, First Faculty of Medicine, Charles University in Prague and General University Hospital in Prague, Prague, Czech Republic. Electronic address:

Background: The efficiency of chimeric antigen receptor (CAR) T-cell-based therapies depends on a sufficient expansion of CAR T cells in vivo and can be weakened by intra-tumoral suppression of CAR T cell functions, leading to a failure of therapy. For example, certain B-cell malignancies such as chronic lymphocytic leukemia are weakly sensitive to treatment with CAR T cells. Co-expression of proinflamatory cytokines such as IL-12 and IL-18 by CAR T cells have been shown to enhance their antitumor function. Read More

View Article and Full-Text PDF
December 2020

Stable integration of an optimized inducible promoter system enables spatiotemporal control of gene expression throughout avian development.

Biol Open 2020 10 6;9(10). Epub 2020 Oct 6.

Department of Orthopaedic Surgery, University of California at San Francisco, 513 Parnassus Avenue, S-1164, San Francisco, CA 94143-0514, USA

Precisely altering gene expression is critical for understanding molecular processes of embryogenesis. Although some tools exist for transgene misexpression in developing chick embryos, we have refined and advanced them by simplifying and optimizing constructs for spatiotemporal control. To maintain expression over the entire course of embryonic development we use an enhanced transposon system that efficiently integrates sequences into the host genome. Read More

View Article and Full-Text PDF
October 2020

CRISPR-mediated Transfection of Brugia malayi.

PLoS Negl Trop Dis 2020 08 31;14(8):e0008627. Epub 2020 Aug 31.

Center for Global Health Infectious Disease Research, College of Public Health, University of South Florida, Tampa, Florida, United States of America.

The application of reverse genetics in the human filarial parasites has lagged due to the difficult biology of these organisms. Recently, we developed a co-culture system that permitted the infective larval stage of Brugia malayi to be transfected and efficiently develop to fecund adults. This was exploited to develop a piggyBac transposon-based toolkit that can be used to produce parasites with transgene sequences stably integrated into the parasite genome. Read More

View Article and Full-Text PDF

PiggyBac mutagenesis and exome sequencing identify genetic driver landscapes and potential therapeutic targets of EGFR-mutant gliomas.

Genome Biol 2020 07 30;21(1):181. Epub 2020 Jul 30.

The Wellcome Trust Sanger Institute, Wellcome Trust Genome Campus, Hinxton, Cambridgeshire, CB10 1SA, UK.

Background: Glioma is the most common intrinsic brain tumor and also occurs in the spinal cord. Activating EGFR mutations are common in IDH1 wild-type gliomas. However, the cooperative partners of EGFR driving gliomagenesis remain poorly understood. Read More

View Article and Full-Text PDF