...company.  Ultragenyx’s goal is to develop novel products for the treatment of rare and ultra-rare diseases...Harvard Medical School, he developed the first lentiviral microRNA-based system for polymerase II-regulated...

...again! Addgenie Marcy (now our Sr. Director of Production) pie-fully celebrating Addgene's 10 year anniversary...ready-to-use viral vectors from some of our most popular lentiviral and AAV vectors, as well as developing and sharing... recombinant antibodies, another plasmid-based product that, with Addgene’s support, could fill a community...

... The “Riboglow” platform for visualizing RNA production and localization Article contributed by Tyler...deaminases like APOBEC. To achieve this they created a lentiviral vector system that monitors both APOBEC- and ...

...genus capable of producing a variety of natural products. Recently, this group optimized their initial ... 10 guide RNAs per gene. The lab modified the lentiviral backbone to encode the CD90.1 selection marker...

...-studied lentivirus is HIV, which many lentiviral tools have been derived from. Lentiviruses have slightly...adenovirus has the highest maximum titer, a plus for production and infection. This virus can infect dividing... with immortalized cell line cultures. Lentivirus Lentivirus is a type of retrovirus and shares many ... commonly used lab viruses: gamma-retrovirus, lentivirus, adenovirus, and adeno-associated virus. Gamma-retrovirus...gamma-retrovirus, coming in around 9 kb. Unlike gamma, lentivirus can infect both dividing and non-dividing cells...nucleus as gamma-retroviruses do. Fun fact: Since lentivirus and gamma-retrovirus are so closely related, ...specific virus? We have you covered with guides on lentivirus, retrovirus, adenovirus, and AAV. If you’re wondering...

...precursors, biofuels, and natural products without the undesirable side products generated during chemical synthesis...these molecules. To make large amounts of pure product in an affordable way, researchers test many different... resistance through gene stacking, increasing production of a protein of interest, and improving the nutrient...Fragmid system was used to assess how different lentiviral plasmid backbone elements impact viral titer ...Modular Assembly Toolkit for the Rapid Design and Production of Diverse Expression Vectors.” ACS Synthetic...expand the Fragmid toolkit to facilitate other lentivirus- and AAV-based studies. Unified MoClo Systems...

...Saccharomyces cerevisiae, as a key host for the production of their products, including fermented beer and wine, ...the authors packaged it into lentivirus and used the resulting lentiviral library to generate a population...Science. 2013. PubMed PMID: 23328393 Wu lab human lentiviral CRISPR library     CRISPR pooled libraries enable...Library. This complex second-generation CRISPR lentiviral library targets nearly all human protein-coding...ingredients, and recombinant proteins. In order to keep production yields high and costs low, and to deal with the...from the CDC and cloned into a third generation lentiviral transfer vector. Each gene has been FLAG-tagged...applications. First, GMAP-compatible backbones for lentivirus LV 1-5 and retrovirus RV 2-5 were constructed...

...., BMC Microbiology 2011 Feb 22;11:38. Lentiviral CRISPR libraries for knockout screening New systems...Pooled Libraries webpage or read our blog post,Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout...CoChR have been deposited to Addgene, including lentiviral and AAV expression vectors for certain variants...purification of Cas9 protein in Rosetta E. coli. New Lentiviral CRISPR activator and repressor plasmids from ...direction of Sabine Suppmann, the Recombinant Protein Production group at Max-Planck Institute of Biochemistry...upon the CRISPR family of plasmids by offering a lentivirus-based mechanism for sgRNA delivery and providing...

...Quality control for Adeno-associated viral vector production. Neuromethods, 195, 77–101. https://doi.org/10.1007...2016). Practical Considerations for Using Pooled Lentiviral CRISPR Libraries. Current Protocols in Molecular...

...risk is high. You are generating a gene therapy product with CRISPR to (hopefully) be used in human clinical...been used - GUIDE-Seq and integrase-defective lentiviral vectors - which rely on insertion of DNA by the...

....7kb, or roughly half the packaging limits of lentiviral and adenoviral vectors. While many transgene ... But despite this, larger functional transgene products were produced following transduction.  So what...

...sites including both the edit rate and the repair product sequence. Targeted amplicon sequencing doesn’t ...CRISPR-Cas9 and TALENs using integrase-defective lentiviral vectors." Nature biotechnology 33.2 (2015): 175... for Your Experiments Get Ready-to-Use CRISPR Lentivirus ...

... with these tools, I found myself writing AAV production tricks into the margins of my protocol. While...life easier!  In this post, I’ll share some AAV production, purification, and titration tips, while also... and analyses needed for packaging AAV. AAV production Overview: The first step in packaging AAV is ...expand the HEK293 cells. Check out the Addgene AAV Production protocol for more details. Pro tips I’m not ... have enough AAV. Compared to the Addgene AAV Production protocol, I take a lazier approach for harvesting...then the viral suspension generated during AAV production is overlaid. Following ultracentrifugation, the...AAV plasmid that was carried over from the AAV production process. Either SYBR green technology or a TaqMan...

...diligently throughout the production process.  Helper-free production Unlike lentivirus, AAV does not possess...For more details on the production process, check out the Addgene AAV Production protocol. Titering Once...experience. Here, we’ll go over the basics of rAAV production.  Of course, you will want to make sure that...facilitate efficient replication. The most common production system, however, is a “helper-free” system, in...).    Figure 1: Overview of key steps of AAV production, iodixanol gradient purification, and AAV titration...remove residual plasmid DNA carried over from the production process. For more details on the qPCR titering... capsid ELISA: Packaging of genomes can limit production of recombinant AAV-2. Gene Therapy, 6(7), Article...

...along with viral packaging plasmids into a viral production cell line, usually HEK-293 cells. When all of...these viral plasmids are expressed within the production cell line, viral particles will be produced and...Typically, one day after transfection into the production cell line (depending on the virus), virus can...Services Resources on the Addgene blog Viral Production at Addgene Viral Vectors 101: What is a Virus?...perform the transduction with: either retrovirus, lentivirus, adenovirus, or AAV. Then, your desired DNA to... some viruses integrate into the host genome (lentivirus and retrovirus) while others are non-integrating...Addgene.org Generating Stable Cell Lines with Lentivirus Virus Handling and Storage Addgene Viral Services...

...iPSCs from dermal fibroblasts via retroviral and lentiviral infections respectively [9, 10]. These stellar...: Implications for Regenerative Medicine and Reproductive Medicine. Geburtshilfe Frauenheilkd, 2015. 75...

...yet a third plasmid (packaging plasmid). Viral production then proceeds by transfection of all three plasmids... into a producer cell line, such as HEK-293T, production of viral particles, and transduction of a target...centrifuging viruses (a necessary step during production), there is a significant risk for aerosol contamination...viruses grown in the lab are gamma retrovirus, lentivirus, adenovirus, and adeno-associated virus (AAV)...humans, earning this virus a BSL-2 classification. Lentivirus (a type of retrovirus) and gamma retrovirus require...and required in BSL-2 labs. When working with lentivirus, many organizations require or recommend disposable...you have to work with BSL-2 materials, such as lentivirus. Most of this work will need to be done in a ...

...Central PMCID: PMC3161101. 20. Chang, C.J., et al., Production of embryonic and fetal-like red blood cells from...that Differentiate the Stem Cell Field Learn how lentiviral vectors can be used for gene delivery Resources...common delivery methods include retrovirus [2], lentivirus [4], adenovirus [5], Sendai virus [6], plasmid...

...What if bacteria snuck into the prep during the production process? These questions are especially important...genome can be packaged inside the vectors during production. These impurities cannot be removed during the...during plasmid growth in bacteria or during viral production. This recombination produces a minor pre-recombined...Sequences during Recombinant Adeno-Associated Virus Production and Their in Vivo Persistence after Vector Delivery....org Find Ready-to-Use AAV Find Ready-to-Use Lentivirus Browse Viral Plasmids ...

...co-infection of helper viruses like adenovirus for productive infection. AAV vectors are engineered to provide... in Addgene's Viral Service Check out our AAV Production Protocol Check out our AAV Purification Protocol...technologies have been investigated including lentiviruses, adenovirus, and inorganic delivery tools such...