We narrowed to 930 results for: Gene
-
TypeBlog PostPublishedJan. 26, 2023, 2:15 p.m....your desired edit are all you need to generate a knock-in. General considerations for designing a repair.... A Mouse Geneticist's practical guide to CRISPR applications. Singh P, et al. Genetics 114.169771; ...accurate mechanism for DSB repair. The process is generally error-free provided the DNA template used for ... DNA to the target sequence to be modified. In general, the insertion sites of the modification should...bases can be difficult, thus dsDNA plasmids are generally preferred for large insertions such as fluorescent...efficiency with circular plasmid templates is generally low; to increase the frequency of edits, researchers...targeting region from the vector. Alternatively, PCR-generated or commercially synthesized linear dsDNA templates...
-
Pooled Library Amplifications
TypeBlog PostPublishedMay 30, 2024, 1:30 p.m....Libraries Resources on Addgene.org Addgene's Guide to Plasmid Pooled Libraries Addgene's Pooled Library Collection...chemical transformations are generally cheaper and easier. One library that Addgene distributes with a chemical...contact us at deposit@addgene.org. Happy amplifying! More resources on the Addgene blog Genome-Wide Screening...If you’re depositing a pooled library with Addgene, you may be surprised to learn that we ask for an ...recombination and loss of plasmid diversity, so Addgene strives to distribute aliquots of the original ...requesting scientist receives a pooled library from Addgene, they’ll need to amplify the sample for further...developed for each individual pooled library; generalized amplification protocols tend not to work as well... -
The Twelve Days of CRISPR
TypeBlog PostPublishedDec. 22, 2020, 2:15 p.m.... CRISPR, Addgene gave to me: a primer on prime editing. On the second day of CRISPR, Addgene gave to ...On the fourth day of CRISPR, Addgene gave to me: ways to use optogenetics to control CRISPR editing. ... day of CRISPR, Addgene gave to me: updates on base editing from the first generation of base editors ...the eighth generation (and dual base editors, too!). On the seventh day of CRISPR, Addgene gave to me... of CRISPR, Addgene gave to me: anti-CRISPR proteins! On the ninth day of CRISPR, Addgene gave to me:...🎶 On the 12 days of CRISPR, Addgene gave to me… 🎶 A few years ago we used the #12DaysOfCRIPSR hashtag...detect nucleic acids. On the third day of CRISPR, Addgene gave to me: tips and tricks for designing gRNAs... -
SpyLigase Irreversibly Locks Peptides Together for Efficient Cell Capture
TypeBlog PostPublishedAug. 13, 2014, 3:49 p.m....no weak links in the chain. Addgene: What about the Streptococcus pyogenes protein domain made it ideal...cover it. Addgene: OK, thanks for your time, and thanks for depositing your plasmids with Addgene. Howarth...lab at the University of Oxford is dedicated to generating new tools to manipulate biology based on molecular...based on a protein isolated from Streptococcus pyogenes that locks itself together, to produce irreversible...another important step forward by dissecting that S. pyogenes protein into three parts. Their efforts yielded... from the “S” in Streptococcus and the “py” in pyogenes), capable of locking two peptide tags together... and its potential uses. Developing a new tool Addgene: Where did you get the idea to develop SpyLigase... -
Plasmids 101: NGS Quality Control for Pooled Libraries
TypeBlog PostPublishedOct. 26, 2017, 1:59 p.m....reference, Addgene commonly prepares NGS samples using both pre- and post-amplification DNA and generates PCR...CRISPR Libraries Resources on Addgene.org Browse Pooled Libraries at Addgene ...In addition to single plasmids, Addgene also distributes pooled plasmid libraries containing hundreds...million plasmids. These libraries are some of Addgene’s most exciting and versatile offerings! We recently...plasmids are amplified and none are lost. Each Addgene library has its own depositor transformation and...components of the library are represented. Typically, Addgene uses electrocompetent Stbl4 cells when performing...transformation efficiency. For a typical library, Addgene usually does four to eight electroporation reactions... -
Celebrating 15 Years of Scientific Sharing
TypeBlog PostPublishedJan. 14, 2019, 2:08 p.m....', style='' %} Resources on the Addgene blog Subscribe to the Addgene blog Read our viral vector blog...blog posts Learn more about Addgene Resources on the Addgene website Find plasmids for your research ...Addgene has so much to celebrate! As we enter this new growth phase of the organization and the expansion...and the opportunities we have for the future. At Addgene we will never ever stop planning, perfecting, and...new projects. Diversification is happening. The Addgene team will have some fun challenges this year and...get excited about making their first deposit to Addgene. We are also lauded by another set of stakeholders...we received some entirely unsolicited praise: “Addgene, for one, is a sterling example of collaboration... -
Building and Validating Recombinant Antibodies: A Case Study
TypeBlog PostPublishedApril 5, 2022, 1:20 p.m....first antibodies distributed by Addgene. The plasmids used to generate these antibodies were based off... used to generate cDNA, which was used as a template for PCR amplification with a degenerate primer set...Introduction to Antibodies Resources on Addgene.org Browse Addgene's recombinant antibody plasmid collection...Addgene has recently announced the launch of an open-access recombinant antibody resource called NABOR...expressed from plasmids that have been deposited with Addgene. Here, we take a closer look at the construction...et. al., Elife A limitation of hybridoma generated mAbs is that if the frozen cell line becomes non-recoverable...grow up the cells. This RNA could then be used to generate functional recombinant monoclonal antibody (R-... -
The Twelve Days of Antibodies
TypeBlog PostPublishedJan. 6, 2025, 5:17 p.m....Antibody, Addgene gave to me… The Addgene Antibody Data Hub! 🎶On the sixth day of Antibody, Addgene gave...Antibody, Addgene gave to me… An Antibodies 101 eBook! 🎶On the second day of Antibody, Addgene gave to...Antibody, Addgene gave to me… An online Antibody Guide! 🎶On the fourth day of Antibody, Addgene gave ...Antibody, Addgene gave to me… Antibodies developed by IPI! 🎶On the eighth day of Antibody, Addgene gave ...Antibody, Addgene gave to me… A recombinant Anti-APP! 🎶On the eleventh day of Antibody, Addgene gave to...Anti-DYKDDDDK! 🎶On the ninth day of Antibody, Addgene gave to me… A neuroscience antibody collection!...flow plots! 🎶On the twelfth day of Antibody, Addgene gave to me… A growing recombinant antibody collection... -
Hot Biosensors 2022: Year-End Roundup
TypeBlog PostPublishedDec. 6, 2022, 2:15 p.m....new to Addgene this year. Feel the love with MTRIA sensors One of the latest additions to Addgene’s catalog...imaging. Vladislav Verkhusha’s lab has generated a new genetically encoded calcium indicator using small...on the Addgene blog Fluorescent Proteins 101: Fluorescent Biosensors Resources on Addgene.org Fluorescent...Here at Addgene’s headquarters, the skies are getting dark and wintry with the end of year approaching...other targets. Figure 1: MTRIA biosensors are generated by inserting a circularly-permuted GFP into an...localized for synaptic glutamate response. They generated variants optimized for different applications ...St-Pierre lab and colleagues have developed a new genetically-encoded voltage indicator (GEVI) called JEDI-... -
Meet an Addgenie: Insights into Grad School, Career Paths, and More
TypeBlog PostPublishedJune 25, 2020, 1:15 p.m....learning.” Addgene’s Director of Product Management Eric Perkins, who has been in many roles in Addgene since...work at Addgene. I posed this question to all nine of my interviewees. Fabienne Furt, Addgene’s Associate...resources on the Addgene blog Browse our science career blog posts Download Addgene's science career guide... guide Resources on Addgene.org Learn more about working at Addgene ...Flynn and I am currently wrapping up my co-op with Addgene as a DNA Sequencing Analyst intern. As a member...go back to school right away. Shannon Rinaldi, Addgene’s Lab Manager, was taking classes at her local community...community college when she took her first job at Addgene as an Order Processing Assistant. After a few months... -
Tips for Technical Support Calls
TypeBlog PostPublishedSept. 20, 2016, 2:39 p.m....Resources on the Addgene Blog Learn About the Addgene Help Center Get Help with the Addgene Deposit Process...emails in my six years as a senior scientist at Addgene. Having spent that long in customer service, I'...Though my experience is based solely on my time at Addgene, I’m confident that these tips and tricks will ...another great source of information. At least at Addgene, the questions in our FAQ section are really, truly...not realize it, all of the reagents available at Addgene come from researchers, and the vast majority of...that for you” meme is infamous enough to have generated it’s own tongue-in-cheek website. If you are ...happens more frequently than you might think. The Addgene scientists answering the phone are smart, but they... -
Year of the Camelid: Antibody Style
TypeBlog PostPublishedMay 9, 2024, 1:15 p.m....from addgene.org Plasmids encoding nanobodies at Addgene Sybody Generation Toolbox from Addgene ...only antibodies. (A) Overview of genetic modifications made to generate the nanomouse. (VHH = variable ...The UN General Assembly has declared 2024 the International Year of Camelids. The declaration is intended...to human populations around the world. Here at Addgene, we love camelids too, though not just for the ...chromobodies,” which are single-domain antibodies genetically fused to fluorescent proteins and used to visualize... synthetic single-domain antibody libraries to generate new sdAbs. There has also been some interesting...) (Xu, et al., 2021). Figure 2: Genetically modified mice offer an alternative method to ... -
Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy
TypeBlog PostPublishedAug. 27, 2020, 1:15 p.m....been generated for yeast (Kim et al., 2019) and fruit flies (Bischof et al., 2018), the generation of such...quantities (and the correct stoichiometric ratios) to generate useable data. What’s more, measuring and quantifying...straightforward nature of BiFC also makes it easier to generate a large number of constructs for high-throughput...screening of PPIs. For example, Bischof et al. (2018) generated an open reading frame (ORF) library to screen ... such resources in less genetically tractable organisms lags behind. BiFC can also be used to screen for...pair. Find BiFC plasmids for your experiment at Addgene! How to Design a BiFC Experiment Much like FRET...optimized to be successful. A BiFC experiment generally involves three steps: fusion protein construction... -
Four Factors that Differentiate the Stem Cell Field
TypeBlog PostPublishedSept. 28, 2016, 2:35 p.m....scientists are generous Besides some of the scientists I've already mentioned, Addgene depositors Marius...What an honor for Addgene to be a part of that! Additional Resources on the Addgene Blog Learn about...landmark Cell paper showing that it was possible to generate induced pluripotent stem cells (iPSCs) capable...called OSKM factors). As the Yamanaka lab's primary Addgene contact for many years, I've been looking forward...example of the fast movement in this field. As an Addgene scientist, I'm exposed to many scientific communities...clinical trial patients with advanced macular degeneration; Laurenz Studer discussed his work with Parkinson's...quality control process alone takes nearly a year. Addgene depositor Juan Carlos Izpsua Belmonte said it took... -
Transferable Skills Guide: Cross-team Communication
TypeBlog PostPublishedNov. 21, 2019, 1:53 p.m....communication blog posts Resources on Addgene.org Find careers at Addgene ... project I’d seen in my already long tenure at Addgene. I would work with our research team and software...pinpointed and (mostly) solved two main problems with Addgene’s cross team communication. Although these real ...these plates now” to a fellow team member. But, at Addgene, we use seven different types of “plates” (Fig....not the container was barcoded (this is pretty Addgene specific) The types or purposes of each given container...that there were a lot of words that nearly all Addgene teams used that seemed simple, but they could in...solution for this problem was the creation of an Addgene lexicon. The base lexicon was and continues to ... -
Three Key Considerations for Precise Neuronal Targeting Using AAV Technologies
TypeBlog PostPublishedNov. 27, 2019, 2:03 p.m....approaches to control transgene expression by restricting expression to genetically defined cell types. ...neuroscientists have begun to use more precise genetic tools: activators, inhibitors, sensors, recombinases...vectors (AAV), neuroscientists can combine classical genetic tools with the manipulation of specific neuronal...changes affect the phenotype. Viral vectors and genetic approaches are continuously refined and developed...molecular tools and sensors Combinations of classical genetic tools, for example Cre/loxP or Flp/FRT, with AAV.... Specific transgene expression can be also achieved in nontransgenic animals by using cis regulatory ... one location can result in expression of an optogenetic payload in neurons that project to different ... -
The 12 Days of CRISPR: 2021
TypeBlog PostPublishedDec. 28, 2021, 2:15 p.m....CRISPR, Addgene gave to me: a primer on prime editing. On the eleventh day of CRISPR, Addgene gave to...all of them here. On the first day of CRISPR, Addgene gave to me: a miniature CRISPR-Cas system for mammalian... mammalian use. On the second day of CRISPR, Addgene gave to me: engineered pegRNAs to improve prime editing...editing efficiency. On the third day of CRISPR, Addgene gave to me: Selective Endogenous eNcapsidation ... Delivery (SEND). On the fourth day of CRISPR, Addgene gave to me: enhanced prime editing systems through...manipulating MMR machinery. On the fifth day of CRISPR, Addgene gave to me: efficient CRISPR editing with a hypercompact...engineered guide RNAs. On the sixth day of CRISPR, Addgene gave to me: PRIME-del (Precise genomic deletions... -
New Viral Vectors - Fall 2024
TypeBlog PostPublishedOct. 3, 2024, 1:15 p.m....preps available in the Addgene viral vector repository! This includes the first Addgene prep packaged in the... AAV1 Optogenetics Karl Deisseroth New serotype pAAV-hSyn-hChR2(H134R)-EYFP AAV2 Optogenetics Karl Deisseroth... Brian O'Neill. More resources on the Addgene blog Addgene's Data Hub What is a Virus (and a Viral Vector...other new AAV vectors as well — including next-generation biosensors for GABA, dopamine, serotonin, and...Table 1: New ready-to-use viral vectors in the Addgene repository Name Serotype Category Depositor...serotype pAAV_hSyn-PdCO-mScarlet-WPRE AAV5 Optogenetics Ofer Yizhar New viral prep pAAV_hSyn-DIO-PdCO-mScarlet-WPRE...-PdCO-mScarlet-WPRE AAV1, AAV5 Optogenetics Ofer Yizhar New viral prep with multiple serotypes pAAV-Syn-FLEX-rc... -
Hot Plasmids - October 2022
TypeBlog PostPublishedOct. 6, 2022, 1:15 p.m....C-terminal region of the human proto-oncogene c-Myc. Since the generation of the first anti-c-Myc [9E10] (...high-throughput screening platform to apply saturating mutagenesis on 21 positions in the protein sequence of the...approach and outcomes of the screen for improved genetically-encoded voltage indicators (GEVIs) based on the...kinetics. They demonstrate the use of HcKCR1 as an optogenetic silencer in experiments with mouse cortical neurons... HcKCR2, making it more suitable for in vivo optogenetic use. Fig. 2: A) HcKCR1 is highly selective...control K+ gradients and can be used as a potent optogenetic silencer. The current work also proposes promising... light-gated potassium channels that mediate optogenetic inhibition. Nat Neurosci 25, 967–974 (2022). ... -
Early Career Researcher Toolbox: Free Online Molecular Biology Tools
TypeBlog PostPublishedSept. 12, 2023, 1:15 p.m....analyze and annotate any DNA sequence. SnapGene Viewer: The free SnapGene Viewer is great for looking at plasmid... biology tools. Addgene Data Hub: A data repository for validation data on Addgene/CLOVER viral vectors...Utah. Figure 1: A plasmid map created in SnapGene of Blue Flame plasmid pET21a-BirA. DNA / .../ protein database search and analysis tools Addgene's analyze sequence tool: Annotates features, BLASTs...restriction enzyme cut sites Open reading frame finder Addgene vector database: Find sequences for plasmids assembled...available sources, as well as those deposited with Addgene. NCBI BLAST: Compares nucleotide or protein sequences...tool, set up an IDT account. Figure 2: Addgene's Analyze Sequence tool. DNA sequence manipulation...