...MTAs per year.  The MTA Process at Addgene As a repository, Addgene doesn’t actually own the majority ...distribution for their plasmids. MTAs at Addgene Since Addgene was founded in 2004, we’ve handled the majority...If you’ve deposited or requested plasmids from Addgene, then you’ve been involved in the material transfer.... The UBMTA, which is the MTA most plasmids at Addgene are shared under, is well-understood and accepted...for managing research use agreements like MTAs.  Addgene does a lot to ensure that we can facilitate a fast...signing of MTAs by the Provider and Recipient (in Addgene parlance, the depositor and requestor) organizations...that differs.  When you deposit a plasmid with Addgene, your organization’s tech transfer office is sent...

...Precious Tools to Improve Gene Modification of Hematopoietic Cells for Research and Gene Therapy. Viruses 12...To deliver genes using lentiviral vectors, you need an envelope protein on the virus’s surface and a ... plasmid which contains a promoter, the envelope gene, and a polyA tail.   Figure 1: Lentiviral...can’t be used for neuronal tracing or peripheral gene delivery. Combining the rabies glycoprotein into...Lentiviral Vectors: One Vector, Many Guises. Human Gene Therapy Methods 28:291–301 . https://doi.org/10.1089...Resources on Addgene.org: Check out the lentivirus guide Find ready-to-use lentiviral preps at Addgene ...your viral vector, you use the same protocol for generating lentivirus, but you use a different envelope ...

...cell lines that stably express a gene of interest. While the general approach for using lentivirus, infect...multiplicity of infection (MOI) for predicting gene transfer events.” Genetic Vaccines and Therapy 2.1 (2004) Pubmed...Mycoplasma infection significantly alters microarray gene expression profiles.” Biotechniques 35.4 (2003)....Downstream processing of oncoretroviral and lentiviral gene therapy vectors.” Biotechnology Advances 24.3 (2006...production of lentiviral and retroviral vectors.” Human Gene Therapy Clinical Development 25.3 (2014). 175-85...lentiviral vectors through pseudotyping.” Current Gene Therapy 5.4 (2005). 387-98. Pubmed PMID: 16101513... cells do not take up nucleic acid or express transgenes as well as actively dividing cells resulting ...

...to knock-out single genes in clonal populations and to knock-out thousands of genes in mixed population...targeting over 19,000 and 20,000 human and mouse genes respectively, use lentiCRISPR v2 as the backbone...Resources on the Addgene Blog Browse Our CRISPR Posts Learn about Using CRISPR to Generate Knockouts Read...announce that the plasmid most requested from the Addgene repository in 2015 was... lentiCRISPR V2   LentiCRISPR...Availability and Use LentiCRISPR v2 is available from Addgene both as a single empty vector and in pooled libraries...reagents available to the research community through Addgene and look forward to another great year of sharing... Plasmids to Mammalian Cells Resources on the Addgene Website Visit our Lentiviral Guide Pages Find ...

... J.A. Engineering the next-generation of CAR T-cells with CRISPR-Cas9 gene editing. Mol Cancer 21, 78 ...dominant cataract disorder caused by a single copy of a gene known as Crygc (Wu et al). They found that the cataracts...able to demonstrate functional correction of the gene in clonally expanded organoids. This work, and many...immunotherapy for cancer treatment by traditional gene therapy pipelines (viral), and researchers are currently...single nucleotide polymorphisms (SNPs) in the TTR gene and is molecularly characterized by misfolding of...blog post on CRISPR’s potential for correcting monogenetic diseases. Now, almost 10 years later, CRISPR’...Resources References Yuxuan Wu et al. "Correction of a Genetic Disease in Mouse Via Use of CRISPR/Cas9." Cell ...

... at Addgene 2. Green, Alexander A., et al. "Toehold switches: de-novo-designed regulators of gene expression...was an in vitro paper-based platform that allowed gene circuits to be deployed out of the laboratory in...combined technologies, we built paper-based synthetic gene networks for the detection of antibiotic resistance...-of-concept work was exciting because it enabled gene circuit-based sensors to be used outside of the ... 3. Pardee, Keith, et al. "Paper-based synthetic gene networks." Cell 159.4 (2014): 940-954. PubMed PMID...Sensor Constructs at Addgene As we report, we have embedded this next generation of toehold switch-based...corresponding portions of the Zika virus genome with Addgene to facilitate rapid sharing of these tools with...

...permanently knock out or knock-in a gene? Do you want to activate or repress gene expression? Are you trying to...reporter gene (e.g. GFP) to identify and enrich positive cells or a selection marker to generate stable ...conjunction with activator or repressor domains to control gene expression. Base editors can help you make precise... separate transfer vectors. May contain reporter gene (e.g. GFP) to identify and enrich positive cells... for each, and potential delivery methods.   General considerations for planning your CRISPR experiment...Wildtype Cas9 or Cas9 nickase are appropriate for generating knockouts or knock-ins, or introducing mutations...PCR followed by gel electrophoresis and/or next-generation sequencing more appropriate? If you already...

...effectively targets your gene/region of interest is a key step for CRISPR/Cas9 gene editing. Dr. Masahito...sequence or gene of interest into the pCoofy vectors, select the appropriate pair of vector and gene primers...vector and gene primer for SLIC cloning. Amplify the pCoofy vector and the sequence of gene of interest... can be used to generate random mutagenesis libraries or to insert heterologous genes, reporters, or other...Updated mini-transposon vector for bacterial mutagenesis or gene targeting Victor de Lorenzo's lab has...approach to genetic engineering which drastically reduces the time and effort needed to insert new genes into...successfully create GFP fusion proteins with a variety of genes across the genome. Although this tool was published...

...Now that we know all about antibiotic resistance genes, let’s consider another basic element of any plasmid...Do you want to make a lot of your plasmid? Is the gene toxic in high amounts? It is always good to keep...below for additional considerations. *Requires pir gene for replication (reference). **F1 is a phage-derived...copies of plasmids if they contain large inserts or genes that create a toxic product. The E. coli strain...survive within cells. The replicons of plasmids are generally different from the those used to replicate the... to make additional copies. ori sequences are generally high in As and Ts. Why, you ask? Well, A-T base...few copies depending on how they are regulated. Generally, control of replication is referred to as "relaxed...

... extensive heterogeneity in the feature sequences due to genetic drift and the use of genes from different...raw plasmid sequence and display frequently used genes and control elements. Development of this tool required...sequence feature that may be used to make fusion genes, detection needs to occur even if one or two codons...of the feature. With the increasing popularity of gene synthesis, many researchers now use codon-optimized... who uses SnapGene or the free SnapGene Viewer companion product. If you don’t use SnapGene yet, you can...Resources on the Addgene Blog Read about our SnapGene powered plasmid maps Learn how we use SnapGene during the...sequencing at Addgene Additional Resources on Addgene.org Example plasmid analysis powered by SnapGene Browse...