A5042666369- CRISPR and Genetic Engineering
- Virus-based gene therapy research
- Cytomegalovirus and herpesvirus research
- Pluripotent Stem Cells Research
- DNA Repair Mechanisms
- Telomeres, Telomerase, and Senescence
- Genetics, Aging, and Longevity in Model Organisms
- Advanced biosensing and bioanalysis techniques
- Microtubule and mitosis dynamics
- Animal Genetics and Reproduction
- Porphyrin Metabolism and Disorders
- Innovation and Socioeconomic Development
- Hemoglobinopathies and Related Disorders
- Viral Infectious Diseases and Gene Expression in Insects
- CAR-T cell therapy research
- Cancer Genomics and Diagnostics
- RNA Interference and Gene Delivery
- RNA and protein synthesis mechanisms
- Mosquito-borne diseases and control
- Genetic and Kidney Cyst Diseases
- Biomedical Research and Pathophysiology
- melanin and skin pigmentation
University of Freiburg
2020-2024
University Medical Center Freiburg
2019-2024
Institute for Transfusion Medicine
2020
Institute of Molecular Biology
2020
Institute of Molecular Biology
2018
University of Applied Sciences Mainz
2018
DKFZ-ZMBH Alliance
2013-2015
Heidelberg University
2013-2015
German Cancer Research Center
2013
Undesired on- and off-target effects of CRISPR-Cas nucleases remain a challenge in genome editing. While the use Cas9 nickases has been shown to minimize mutagenesis, their therapeutic editing hampered by lack efficacy. To overcome this limitation, we others have developed double-nickase-based strategies generate staggered DNA double-strand breaks mediate gene disruption or correction with high efficiency. However, impact paired single-strand nicks on integrity remained largely unexplored....
The therapeutic use of adeno-associated viral vector (AAV)-mediated gene disruption using CRISPR-Cas9 is limited by potential off-target modifications and the risk uncontrolled integration genomes into CRISPR-mediated double-strand breaks. To address these concerns, we explored AAV-delivered paired Staphylococcus aureus nickases (D10ASaCas9) to target Hao1 for treatment primary hyperoxaluria type 1 (PH1). Our study demonstrated effective disruption, a significant decrease in glycolate...
Abstract Genome editing by homology directed repair (HDR) is leveraged to precisely modify the genome of therapeutically relevant hematopoietic stem and progenitor cells (HSPCs). Here, we present a new approach increasing frequency HDR in human HSPCs delivery an inhibitor 53BP1 (named “i53”) as recombinant peptide. We show that use i53 peptide effectively increases HDR-mediated at variety loci well other primary cell types. incorporating protein allows high frequencies while lowering amounts...
Abstract The DNA-PKcs inhibitor AZD7648 enhances CRISPR–Cas9-directed homology-directed repair efficiencies, with potential for clinical utility, but its possible on-target consequences are unknown. We found that genome editing causes frequent kilobase-scale and megabase-scale deletions, chromosome arm loss translocations. These large-scale chromosomal alterations evade detection through typical assays, prompting caution in deploying reinforcing the need to investigate multiple types of outcomes.
Abstract Precise genome editing requires the resolution of nuclease-induced DNA double strand breaks (DSBs) via homology-directed repair (HDR) pathway. In mammals, this is typically outcompeted by non-homologous end-joining (NHEJ) that can generate potentially genotoxic insertion/deletion mutations at DSB sites. Because higher efficacy, clinical has been restricted to imperfect but efficient NHEJ-based approaches. Hence, strategies promote HDR are essential facilitate transition HDR-based...
The programmable nuclease technology CRISPR-Cas9 has revolutionized gene editing in the last decade. Due to risk of off-target editing, accurate and sensitive methods for characterization are crucial prior applying therapeutically. Here, we utilized a rhesus macaque model compare predictive values CIRCLE-seq, an vitro prediction method, with silico (ISP) based solely on genomic sequence comparisons. We use AmpliSeq HD error-corrected sequencing validate sites predicted by CIRCLE-seq ISP CD33...
Cells challenged with DNA damage activate checkpoints to arrest the cell cycle and allow time for repair. Successful repair coupled subsequent checkpoint inactivation is referred as recovery. When cannot be repaired, a choice between permanent cycling in presence of (checkpoint adaptation) must made. While jeopardizes future lineages, continued proliferation associated risk genome instability. We demonstrate that nutritional signaling through target rapamycin complex 1 (TORC1) influences...
Aging | doi:10.18632/aging.100835. Clemens Stockklausner, Simon Raffel, Julia Klermund, Obul Reddy Bandapalli, Fabian Beier, Tim H. Brümmendorf, Friederike Bürger, Sven W. Sauer, Georg F. Hoffmann, Holger Lorenz, Laura Tagliaferri, Daniel Nowak, Wolf-Karsten Hofmann, Rebecca Buergermeister, Carolin Kerber, Tobias Rausch, Jan O. Korbel, Brian Luke, Andreas Trumpp, E. Kulozik
Genome editing with programmable nucleases has shown great promise for clinical translation but also revealed the risk of genotoxicity caused by chromosomal translocations or insertion mutations at off-target sites. Here, we describe CAST-Seq, an innovative assay to identify and quantify aberrations derived from on- activities CRISPR-Cas TALENs. CAST-Seq detected novel types rearrangements, including homology-mediated that are mediated homologous recombination. Depending on employed designer...
Abstract Human cancers frequently harbour mutations in DNA repair genes, rendering the use of damaging agents as an effective therapeutic intervention. As therapy-resistant cells often arise, it is important to better understand molecular pathways that drive resistance order facilitate eventual targeting such processes. We employ repair-defective diploid yeast a model demonstrate that, response genotoxic challenges, nearly all eventually undergo checkpoint adaptation, resulting generation...
Human cancers frequently harbour mutations in DNA repair genes, rendering the use of damaging agents as an effective therapeutic intervention. As therapy-resistant cells often arise, it is important to better understand molecular pathways that drive resistance order facilitate eventual targeting such processes. We employ recombination-defective diploid yeast a model demonstrate that, response genotoxic challenges, nearly all eventually undergo checkpoint adaptation, resulting generation...
ABSTRACT The programmable nuclease technology CRISPR/Cas9 has revolutionized gene editing in the last decade. Due to risk of off-target editing, accurate and sensitive methods for characterization are crucial prior applying therapeutically. Here, we utilized a rhesus macaque model ask whether CIRCLE-Seq (CS), an vitro prediction method, more accurately identifies off-targets compared silico (ISP) based solely on genomic sequence comparisons. We use AmpliSeq HD error-corrected sequencing...