A5088568460- CRISPR and Genetic Engineering
- Virus-based gene therapy research
- CAR-T cell therapy research
- Pluripotent Stem Cells Research
- Viral Infections and Immunology Research
- Viral Infectious Diseases and Gene Expression in Insects
- Cytomegalovirus and herpesvirus research
- Genetics, Aging, and Longevity in Model Organisms
- Advanced biosensing and bioanalysis techniques
- Porphyrin Metabolism and Disorders
- Innovation and Socioeconomic Development
- RNA Interference and Gene Delivery
- Immune Cell Function and Interaction
- HIV Research and Treatment
University of Freiburg
2020-2024
University Medical Center Freiburg
2020-2024
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...
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...
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...
Liver-directed adeno-associated viral (AAV) vector-mediated homology-independent targeted integration (AAV-HITI) by CRISPR-Cas9 at the highly transcribed albumin locus is under investigation to provide sustained transgene expression following neonatal treatment. We show that targeting 3′ end of results in productive about 15% mouse hepatocytes achieving therapeutic levels systemic proteins two models inherited diseases. demonstrate full-length HITI donor DNA preferentially integrated upon...
X-linked lymphoproliferative disease is a rare inherited immune disorder, caused by mutations or deletions in the SH2D1A gene that encodes an intracellular adapter protein SAP (Slam-associated protein). essential for mediating several key processes and system - T cells particular are dysregulated its absence. Patients present with spectrum of clinical manifestations, including haemophagocytic lymphohistiocytosis (HLH), dysgammaglobulinemia, lymphoma autoimmunity. Treatment options limited,...
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...
Persistent hepatitis B virus (HBV) infection remains a serious medical problem worldwide, with an estimated global burden of 257 million carriers. Prophylactic and therapeutic interventions, in the form vaccine, immunomodulators, nucleotide nucleoside analogs, are available. Vaccination, however, offers no benefit to chronic sufferers has had limited impact on rates. Although immunomodulators analogs have been licensed for treatment HBV, cure rates remain low. Transcription activator-like...
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...
<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...