A5047652907- CAR-T cell therapy research
- CRISPR and Genetic Engineering
- Virus-based gene therapy research
- Viral Infectious Diseases and Gene Expression in Insects
- Immune Cell Function and Interaction
- RNA Interference and Gene Delivery
- Advanced biosensing and bioanalysis techniques
- Viral Infections and Immunology Research
- Pluripotent Stem Cells Research
- Silicon Carbide Semiconductor Technologies
- T-cell and B-cell Immunology
- Genetics, Aging, and Longevity in Model Organisms
- Immune cells in cancer
- Extracellular vesicles in disease
- Immune Response and Inflammation
- Immunotherapy and Immune Responses
- Innovation and Socioeconomic Development
- Advancements in Semiconductor Devices and Circuit Design
- RNA regulation and disease
University Medical Center Freiburg
2013-2024
University of Freiburg
2017-2024
Medizinische Hochschule Hannover
2014-2015
Designer nucleases have been successfully employed to modify the genomes of various model organisms and human cell types. While specificity zinc-finger (ZFNs) RNA-guided endonucleases has assessed some extent, little data are available for transcription activator-like effector-based (TALENs). Here, we engineered TALEN pairs targeting three loci (CCR5, AAVS1 IL2RG) performed a detailed analysis their activity, toxicity specificity. The TALENs showed comparable activity benchmark ZFNs, with...
Natural Killer (NK) cells are known for their high intrinsic cytotoxic capacity, and the possibility to be applied as 'off-the-shelf' product makes them highly attractive cell-based immunotherapies. In patients with multiple myeloma (MM), an elevated number of NK has been correlated higher overall-survival rate. However, cell function can impaired by upregulation inhibitory receptors, such immune checkpoint NKG2A. Here, we developed a CRISPR-Cas9-based gene editing protocol that allowed us...
The potential of adoptive cell therapy can be extended when combined with genome editing. However, variation in the quality starting material and different manufacturing steps are associated production failure product contamination. Here, we present an automated T engineering process to produce off-the-shelf chimeric antigen receptor (CAR) cells on CliniMACS Prodigy platform containing in-line electroporation unit. This setup was used combine lentiviral delivery a CD19-targeting CAR transfer...
While chimeric antigen receptor (CAR) T cell immunotherapy targeting CD19 has shown remarkable success in patients with lymphoid malignancies, the potency of CAR cells solid tumors is low so far. To improve efficacy prostate carcinoma, we designed a novel that recognizes new epitope prostate-specific membrane (PSMA) and established paradigms to apply preclinical cancer model. In vitro characterization D7 single-chain antibody fragment-derived anti-PSMA confirmed choice co-stimulatory domain...
Abstract Efficient delivery of genetic material to primary cells remains challenging. Here, efficient transfer is presented using synthetic biodegradable nanocarriers, resembling extracellular vesicles in their biomechanical properties. This based on two main technological achievements: generation soft polyelectrolyte capsules nanosize and application the nanocapsules for co‐transfer different RNAs tumor cell lines cells, including hematopoietic progenitor T cells. Near 100% efficiency...
Designer nucleases, like zinc-finger nucleases (ZFNs), represent valuable tools for targeted genome editing. Here, we took advantage of the gamma-retroviral life cycle and produced vectors to transfer ZFNs in form protein, mRNA episomal DNA. Transfer efficacy ZFN activity were assessed quantitative proof-of-concept experiments a human cell line mouse embryonic stem cells. We demonstrate that retrovirus-mediated protein (RPT), (RMT) episome (RET) powerful methodologies transient delivery or...
While chimeric antigen receptor (CAR) T cell therapy has shown promising outcomes among patients with hematologic malignancies, it also been associated undesirable side-effects such as cytokine release syndrome (CRS). CRS is triggered by CAR T-cell-based activation of monocytes, which are stimulated via the CD40L–CD40R axis or uptake GM-CSF to secrete proinflammatory cytokines. Mouse models have used model CRS, but working them labor-intensive and they not amenable screening approaches. To...
Zinc finger nucleases (ZFNs) consist of zinc fingers as DNA-binding module and the non-specific DNA-cleavage domain restriction endonuclease FokI module. This architecture is also used by TALE (TALENs), in which modules ZFNs have been replaced domains based on transcription activator like effector (TALE) proteins. Both TALENs are programmable rely dimerization to induce double-strand DNA cleavage at target site after recognition respective seem an advantage over ZFNs, assembly proteins...
In vitro disease modeling based on induced pluripotent stem cells (iPSCs) provides a powerful system to study cellular pathophysiology, especially in combination with targeted genome editing and protocols differentiate iPSCs into affected cell types. this study, we established zinc-finger nuclease-mediated primary fibroblasts generated from mouse model for radiosensitive severe combined immunodeficiency (RS-SCID), rare disorder characterized by sensitivity radiation the absence of...
Abstract Chimeric antigen receptor (CAR) T cell technology has enabled successfully novel concepts to treat cancer patients, with substantial remission rates in lymphoid malignancies. This therapy is based on autologous lymphocytes that are genetically modified express a CAR recognizes tumor-associated antigens and mediates the elimination of respective tumor cells. Current limitations include laborious manufacturing procedures as well severe immunological side effects upon administration To...
Inherited defects in MyD88 and IRAK4, two regulators Toll-like receptor (TLR) signaling, are clinically highly relevant, but still incompletely understood. MyD88- IRAK4-deficient patients exceedingly susceptible to a narrow spectrum of pathogens, with ∼50% lethality the first years life. To better understand underlying molecular cellular characteristics that determine disease progression, we aimed at modeling response pathogens vitro . this end, determined immunophenotype monocytes...
The generation of T cells from pluripotent stem (PSCs) is attractive for investigating cell development and validating genome editing strategies in vitro. X-linked severe combined immunodeficiency (X-SCID) an immune disorder caused by mutations the IL2RG gene characterised absence NK patients. encodes common gamma chain, which part several interleukin receptors, including IL-2 IL-7 receptors. To model X-SCID vitro, we generated a mouse embryonic (ESC) line disease-causing human variant...
The CRISPR-Cas12a platform has attracted interest in the genome editing community because prototypical Acidaminococcus Cas12a generates a staggered DNA double-strand break upon binding to an AT-rich protospacer-adjacent motif (PAM, 5'-TTTV). broad application of primary human cells was enabled by development engineered version natural protein, called Ultra. In this study, we confirmed that Ultra ribonucleoprotein complexes allelic gene disruption frequencies over 90% at multiple target sites...
Topic: 24. Gene therapy, cellular immunotherapy and vaccination - Biology & Translational Research Background: CD33-targeting chimeric antigen receptor (CAR; CAR33)-T cells already showed efficacy for the treatment of acute myeloid leukemia (AML). Yet clinical application CAR33-T remains challenging due to its restriction autologous cell preparations potential risk severe side effects. In contrast, natural killer (NK) can be safely administered HLA-mismatched recipients possess an intrinsic...
<title>Abstract</title> Background The development of the CRISPR-Cas12a platform has generated considerable interest in genome editing community. Due to its AT-rich protospacer-adjacent motif (PAM, 5’-TTTV), Cas12a increased potential number targetable sites for gene beyond that prototypical<italic>Streptococcus pyogenes</italic>CRISPR-Cas9 system. Moreover, evaluation off-target activity nucleases suggested high specificity platform. Broad application primary human cells was recently...