Laverock Therapeutics’ Post

#ESGCT poster presented by @NicolasMartin on our FlashRNA® technology, as a safe and efficient solution for RNA transfer into organoids, in vivo models, iPSCs and stem cells. 🔑 Key points: - RNA transfer without risk of cellular alteration. - Applicable to organoids, iPSCs and in vivo approaches - Ideal for genome editing, regenerative medicine and immuno-oncology Our collaboration with Urosphere demonstrates the efficacy of FlashRNA® on cancerous bladder organoids. Want to find out more about this technology, which is revolutionizing cell and gene therapy? Click here: https://lnkd.in/d-S9q8Uk #GeneTherapy #Bioproduction #Biotechnology #FlashRNA #Organoids European Society of Gene and Cell Therapy Urosphere

#biology #molecularbiology #gene #geneediting #bioinformatics Regulatory considerations in CAR-T Cell treatment After the approval of the first chimeric antigen receptor T cell (CAR-T) called Tisagenlecleucel (a drug for the treatment of B-cell acute lymphoblastic leukemia (ALL)) by the FDA, the use of genome-edited human cells in treatment has increased dramatically. . Advancing success in this treatment method requires cooperation between researchers and regulatory authorities. In this regard, the #FDA has recently published two official guidelines related to human gene therapy and cell therapy products, which include recommendations on gene editing in somatic cells and considerations for the development of CAR-T cells.  👉Genome editing based on the genomic target, editing system used and factors inside the host cell (such as natural human genetic diversity) can have different efficiency. #Homology-based editing and non-homologous end joining can lead to rearrangements, aneuploidy, and off-target mutations. These possibilities raise the importance of concern about the efficacy and safety of this treatment, which has led to further FDA recommendations for genomic testing of such products before use in patients.  Evaluation of products for off-target mutations is often done with approaches in preclinical and clinical stages. Preclinical assessments using labeling of DNA double-strand break sites are likely to miss structural rearrangements and aneuploidy, and these strategies cannot be applied directly to CAR T-cells administered to patients. As a result, engineered clinical products are often tested through more conventional technologies, including #FISH and G-banded karyotyping, to screen for genome integrity and analyze off-target mutations in selected regions via PCR with specific primers, which approach also Requires live cell or development of assays specific to each cell product.  👉The lack of an efficient and comprehensive approach to CAR T-Cell evaluation is an obstacle in the clinical trial evaluation of these products. The use of whole genome sequencing (WGS: Whole genome sequencing) is recommended for genomic analysis of engineered T #cells in patients. Among the advantages of this approach compared to conventional molecular tests, we can mention the use of #DNA as input instead of living cells, the ability to simultaneously evaluate the efficiency of on-target editing and screening of different types of off-target mutations, and to specify the sites of gene insertion.  It is hoped that in the future, the safety profiles of CAR T-cell #therapies will be established through detailed tests with highly sensitive #laboratory assays during clinical development, and WGS will be performed on the clinical product as a final quality control before delivery to the patient. More information: https://lnkd.in/dci6mwnY

How do we overcome the limitation for the size of gene of interest with AAV (4.7 kb? By introduce dual AAVs with mechanism of mRNA trans-splicing, it has shown successfully in both preclinical and clinical settings. mRNA trans-splicing dual AAV vectors for (epi)genome editing and gene therapy Large genes including several CRISPR-Cas modules like gene activators (CRISPRa) require dual adeno-associated viral (AAV) vectors for an efficient in vivo delivery and expression. Current dual AAV vector approaches have important limitations, e.g., low reconstitution efficiency, production of alien proteins, or low flexibility in split site selection. Here, we present a dual AAV vector technology based on reconstitution via mRNA trans-splicing (REVeRT). REVeRT is flexible in split site selection and can efficiently reconstitute different split genes in numerous in vitro models, in human organoids, and in vivo. Furthermore, REVeRT can functionally reconstitute a CRISPRa module targeting genes in various mouse tissues and organs in single or multiplexed approaches upon different routes of administration. Finally, REVeRT enabled the reconstitution of full-length ABCA4 after intravitreal injection in a mouse model of Stargardt disease. Due to its flexibility and efficiency REVeRT harbors great potential for basic research and clinical applications.

Introducing CD Genomics' AAV Integration Site Analysis Service! Adeno-Associated Virus (AAV) vectors are revolutionizing gene therapy, but understanding their integration into the host genome is crucial for optimizing safety and efficacy. Our advanced AAV Integration Site Analysis service provides comprehensive insights into AAV genome behavior post-administration, enabling: 🔬 Detailed characterization of integration sites 🛡️ Enhanced safety assessment to mitigate oncogenic risks 🎯 Optimization of gene therapy strategies 📊 Support for regulatory compliance Using cutting-edge techniques like hybrid-capture-based target enrichment sequencing and custom bioinformatics pipelines, we offer unparalleled insights into AAV integration patterns—critical for applications in gene therapy, oncology, immunology, and preclinical studies. Explore how our service can elevate your research and therapeutic development! 👉 Learn More: https://lnkd.in/gZBrjYKe #GeneTherapy #AAV #Biotechnology #Genomics

🔬 Unlocking Disease Mechanisms with Gene Knockout and Knock-In Models 🧬 In our most recent article we explore the powerful applications of gene knockout and knock-in models in understanding and treating various diseases. Key Highlights: -Innovative Tools: Discover the latest in gene editing technologies, including CRISPR/Cas9, TALENs, and ZFNs. -Understanding Diseases: How these models help decode gene functions and disease mechanisms. -Therapeutic Development: The role of gene editing in developing cutting-edge treatments like CAR T-cell therapy. -Challenges and Future Directions: Insights into the limitations and potential of these models in disease research. These technologies are advancing our knowledge of diseases like cancer, cardiovascular conditions, and neurodegenerative disorders and paving the way for novel therapeutic strategies. 🔗Read the article here: https://lnkd.in/eyJSthUv #GeneEditing #CRISPR #BiomedicalResearch #DiseaseMechanisms #TherapeuticDevelopment #GeneticResearch #Biotechnology #HealthInnovation #PrecisionMedicine

With the proven success of in vivo genome editing, it will open a new door for treating many diseases, one injection, lifelong effectiveness, safety profile of #AAV In vivo genome editing via CRISPR/Cas9-mediated homology-independent targeted integration for Bietti crystalline corneoretinal dystrophy treatment Bietti crystalline corneoretinal dystrophy (BCD) is an autosomal recessive chorioretinal degenerative disease without approved therapeutic drugs. It is caused by mutations in CYP4V2 gene, and about 80% of BCD patients carry mutations in exon 7 to 11. Here, we apply CRISPR/Cas9 mediated homology-independent targeted integration (HITI)-based gene editing therapy in HEK293T cells, BCD patient derived iPSCs, and humanized Cyp4v3 mouse model (h-Cyp4v3mut/mut) using two rAAV2/8 vectors via sub-retinal administration. We find that sgRNA-guided Cas9 generates double-strand cleavage on intron 6 of the CYP4V2 gene, and the HITI donor inserts the carried sequence, part of intron 6, exon 7-11, and a stop codon into the DNA break, achieving precise integration, effective transcription and translation both in vitro and in vivo. HITI-based editing restores the viability of iPSC-RPE cells from BCD patient, improves the morphology, number and metabolism of RPE and photoreceptors in h-Cyp4v3mut/mut mice. These results suggest that HITI-based editing could be a promising therapeutic strategy for those BCD patients carrying mutations in exon 7 to 11, and one injection will achieve lifelong effectiveness. https://lnkd.in/e_8u2MkX

Researchers at Aarhus University have developed a groundbreaking "trimodal genetic engineering" system using orthogonal CRISPR/Cas technologies. This method allows simultaneous activation, repression, and deletion of genes within a single cell by combining dSpCas9-VPR for activation, dSaCas9-KOX1 for repression, and AsCas12a for deletion. Optimized in Jurkat cells and primary human T cells, the approach achieved precise, transient gene regulation alongside permanent modifications with minimal off-target effects. Demonstrating efficiency in applications like CAR T-cell therapy, the system holds immense potential for cell therapy and regenerative medicine, enabling complex gene interactions crucial for treating diseases with intricate genetic networks. https://lnkd.in/dDDqkhnr

🔥 Still excited about this Cell & Gene Therapy news from last month! 🔥 About a month ago, Prime Medicine and Bristol Myers Squibb (BMS) announced a $55M collaboration to develop next-gen T-cell therapies using Prime’s innovative PASSIGE™ technology for large-scale, non-viral gene editing. Even though it’s been a few weeks, I’m still thinking about how this could be a game-changer for cancer and immune-related diseases. As someone who’s spent years in oncology and cell therapy, I’m excited about the potential here. The ability to scale precise, non-viral gene editing could really reshape how we approach personalised medicine, especially for patients with high unmet needs. Key takeaways: • $55M investment to drive innovation in gene editing. • Focus on developing T-cell therapies for cancer and immunological diseases. • Non-viral, large-scale gene editing could revolutionize treatment approaches. Even though this news is a few weeks old, it’s still worth discussing because of its potential impact on the future of #CellTherapy and #GeneTherapy. What do you think? Let’s chat! #Oncology #Biotech #Innovation #CGT Source: https://lnkd.in/eqTRcKYw

🔬 Excited to share that our latest research paper, "Temporal Insights into Molecular and Cellular Responses during rAAV Production in HEK293T Cells," has been published! 🧬 🔍This study analyzes various transfection conditions for the production of recombinant adeno-associated viruses (rAAVs) in HEK293T cells using SWATH-mass spectrometry. We have identified important proteins and pathways that can help boost rAAV yield, optimize transfection, and improve gene therapy vector production. 🏭 Industry Impact: This research holds immense promise for the biomanufacturing sector, offering valuable insights into cellular responses during rAAV production. By deciphering the molecular intricacies of gene therapy vector manufacturing, we aim to drive advancements in production processes and elevate the efficiency of rAAV production for diverse therapeutic applications. Please read the full paper to learn more about our contributions to this critical field. 𝐡𝐭𝐭𝐩𝐬://𝐰𝐰𝐰.𝐜𝐞𝐥𝐥.𝐜𝐨𝐦/𝐦𝐨𝐥𝐞𝐜𝐮𝐥𝐚𝐫-𝐭𝐡𝐞𝐫𝐚𝐩𝐲-𝐟𝐚𝐦𝐢𝐥𝐲/𝐦𝐞𝐭𝐡𝐨𝐝𝐬/𝐟𝐮𝐥𝐥𝐭𝐞𝐱𝐭/𝐒2329-0501(24)00094-9 #Research #GeneTherapy #Biomanufacturing #Proteomics #rAAV #HEK293T#A*STAR Bioprocessing Technology Institute (BTI)#A*STAR

Excited to share groundbreaking research on using pseudotyped lentivirus-derived nanoparticles (LVNPs) for targeted delivery of CRISPR-Cas9 ribonucleoprotein complexes. This modular approach enables precise gene editing in specific cell populations, including human primary B cells, achieving indel rates exceeding 80%. The study highlights the versatility of engineered viral glycoproteins from SARS-CoV-2, Nipah virus, and measles virus, offering tailored solutions for therapeutic applications, from autoimmune diseases to cancer. This innovation paves the way for safer and more effective genome editing therapies. Kudos to the research team for this leap forward in molecular medicine! Curious? Explore more here: https://lnkd.in/dT5BHmk3 #GeneTherapy #CRISPR #InnovationInScience #BiomedicalResearch