We narrowed to 932 results for: Gene

...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...

... 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...

...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...

...containing cassettes of the same gene multiple times.  Bioinformatics at Addgene After a run on our Illumina...Resources on the Addgene Blog A Look at Addgene’s QC Process Bioinformatics at Addgene The Power Behind...Previously, we provided a general overview of the QC process at Addgene. All plasmids go through this...technologies that Addgene uses in the QC process, when these technologies are used, and how Addgene keeps up with...Sequencing technologies used at Addgene Sanger sequencing (“First-generation sequencing”)  Sanger sequencing...assembles the raw data.  Addgene’s de novo assembly pipeline usually generates the correct plasmid sequence...revisit the assembly in a program called Geneious. Within Geneious, the raw FASTQ reads are used for either...

...plasmid to ensure our assembly result is accurate The gene/insert or any major feature highlighted by the deposited...sequenced by Addgene’s QC team. Email help@addgene.org to get started! Addgene’s QC process Addgene prioritizes...deposited sequence(s) in SnapGene. SnapGene annotates the plasmid maps based on SnapGene’s “Common Feature” library...Quality Control Now at Addgene Bioinformatics at Addgene Resources on Addgene.org Educational Resources... Figure 2. Addgene’s basic QC process. Created with BioRender.com. Once Addgene receives deposited...receiving the replacement plasmid at Addgene. QC is a team effort Addgene’s QC team is full of dedicated scientists... Resources Additional resources on the Addgene blog Addgene's Tips for Plasmid Quality Control Plasmids...

...do scroll down? You’ll find Blugene, as well as information about Addgene, our mission, links to our deposit...improving the tools you use to share and access them. Addgene’s product, development, and content teams have made... made some changes to the look and feel of addgene.org. Some you’ll notice right away, while others may...resources, tools, collections in our catalog, and the Addgene Help Center. A big part of this redesign was pulling... place, so you can quickly navigate around the Addgene website without scrolling up and down the page....skim through. Users with disabilities will find Addgene’s website easier to navigate, thanks to design choices...— check out all the new features and designs addgene.org! And when you’re finished, feel free to take ...

... plasmid encoding rep and serotype-specific cap genes, and a plasmid encoding adenoviral helper sequences...viral vectors are tested in vitro and in vivo for gene expression and/or function. These data are sometimes...copies of the lentiviral Rev responsive element (RRE) gene and the cellular ribonuclease P/MRP 30 kDa subunit...vector genome titers by droplet digital PCR. Human Gene Therapy Methods, 25(2), Article 2. https://doi.org...Titer by a Novel Droplet Digital PCR Method. Human Gene Therapy Methods, 29(2), 96–103. https://doi.org/...Production AAV distributed by Addgene has been produced either in-house by Addgene scientists or through collaboration...either performed by Addgene or by the University of Pennsylvania Vector Core. In general, titering is performed...

...means summer is right around the corner. Here at Addgene, we’re always inspired by the new seasons, so we've...complex questions, and giving tips for consistently generating solid data. If you’re ready for the next steps...We’re very excited to be launching the official Addgene TikTok at the end of this month! We’ll be sharing...lab, along with some behind-the-scenes peaks of Addgene’s people, culture, and workplace. Rumor has it that...

...mail due to laws concerning genetically modified organisms. Since Addgene is very proud to help scientists...times easier. We’ve certainly felt this way at Addgene so we’ve been putting some work into finding robots... many more robots than you ever knew existed! Addgene recently started automating a few protocols to ... hour. Automating vial fiIling The majority of Addgene plasmids are shipped as agar stabs in glass vials... easily programmed for a variety of lab needs. Addgene uses it to fill as many vials as it can in the ...Label your tubes with a label maker. Label makers generate far more legible labels than your hand and help...avoid sore fingers. There are other process that Addgene would love to move forward and automate. For instance...

...targeting EvolvR to five dispersed regions of the target gene of spectinomycin, rpsE. These results not only uncovered...directed mutagenesis to generate specific mutations in your plasmids Use REPLACER Mutagenesis to mutate...and forward genetics harnesses this at a large scale by screening diverse libraries of genetic variants....Berkeley, provides a new approach for generating novel genetic variants in bacteria. Read on to learn about...EvolvR’s on-target mutagenesis rate was 2.5 x 10-6 mutations per nucleotide per generation vs. 10 x 10-10 ...additional mutations to PolI3M generated PolI5M which increased EvolvR’s mutagenesis rate to ~10-3 mutations ...than one genomic site for mutagenesis, a feature which they used to generate bacteria resistant to both...

...Halloween is only 2 days away and here at Addgene we couldn’t be more excited! As a small, tight-knit...email system.  Our 2013 winners dressed up as Addgene's favorite lunch... a BonMe noodle meal, crammed...interested in finding out who wins this year’s Addgene costume contest, like us on Facebook and checkout...  addgene...

...-invasive systemic gene delivery in non-human primate (NHP) brains.  Systemic gene delivery capsids previously...Monomeric StayGold variants AAV.CAP-Mac for systemic gene transfer in primate brains Endothelial-specific ...Adeno-associated viral vectors for functional intravenous gene transfer throughout the non-human primate brain....plasmid here! Chen, X., et al. (2023). Functional gene delivery to and across brain vasculature of systemic...past decade, primary cells remain difficult to genetically edit due to toxicity from electroporation or ... under CC-BY license.   The Miyawaki lab generated a series of StayGold mutants and identified a ...mentioning that Kiryl Piatkevich and colleagues generated yet another monomeric StayGold variant (Piatkevich...

...these expected hits, we always get hits to “random” genes. Do these hits always mean the sample is contaminated...matters is the number of hits to the same sequence (gene). In a clean sample, there are typically fewer than... However, a large number of hits to one sequence/gene - we set our threshold at >100 hits - immediately...example, AAV-68544 contained 220 hits to the CHRM4 gene. When contamination is suspected, we proceed to...Viral Preparations from Viral DNA Extracts. Human Gene Therapy 31:664–678 . https://doi.org/10.1089/hum...purity. But Addgene is also conducting more rigorous quality control, including next generation sequencing... for transgene expression. Once packaged in AAV, this pre-recombined DNA could lead to transgene expression...

...her work has always been focused on gene transfer, from transgenes to plants, chloroplast to the nucleus...easy to express genes in, which makes it a good laboratory workhorse. Tips & tricks Addgene: Do you have ...The MoClo Plant Kit makes it really easy to clone genes for expression in plant cells. Sylvester and I chose... Now there are many, so if you want to make a 10-gene construct, you don’t have to worry about repeating...constructs to allow people to, for example, knock out a gene with simultaneous complementation, with a knockout...Marillonnet share with the scientific community via Addgene, the struggles of plant scientists and how they...Everybody's favorite parts for plant synthetic biology Addgene: Tell me about the Golden Gate Modular Cloning ...

...from that of the gene of interest, or expressed from the same transcript as the gene of interest, but ...visualize when and where the gene is expressed. Click here to view Addgene’s collection of empty backbones... a wide number of functions, including: tagging genes for elucidating their expression or localization...promoter of interest can be used to effectively monitor gene expression from that promoter in a given cell type...more about optogenetics at OpenOptogenetics and find optogeneic actuators and sensors at Addgene. Cell marking...and others. Review Addgene's collection of fluorescent biosensors here. Optogenetics: Scientists can use...implantation and generation of transgenic mice. Purification: GFP can be used as a general epitope tag for...

...dealing with a defective gene, Xm, the sequence of which is identical to the correct gene, Xw, except for a ...Adenine base editors (ABE) What if you have another gene Y, which has a defective allele Ym and you need ...first created a defective chloramphenicol resistance gene (CamR) by introducing a point mutation (H193Y). ...transformed it into E. coli harboring the defective CamR gene. Screening for surviving colonies and subsequent...system. We use a single polycistronic tRNA-gRNA (PTG) gene system to clone multiple protospacers which enables... slim chance of generating indels with base editors, keep in mind that the generation of indel sometimes...biology Resources on Addgene.org Find all CRISPR plasmids and resources at Addgene Find CRISPR base editors...