...of highly active sgRNAs for CRISPR-Cas9-mediated gene inactivation." Nature biotechnology (2014). PubMed...to select a tool based on your project and not general opinion. I could spend a lot of time running through... Online tools dominate the software space but generally rely on less powerful algorithms to detect off-targets...tools available mostly work on any computer but generally require working knowledge of the command line ...tools require some kind of data input in order to generate results. While different types of input are more...focus on specific organisms. For this reason I generally recommend that laboratories that have already ...PubMed Central PMCID: PMC4262738. Resources at Addgene Get Tips on How to Design a gRNA Read Our CRISPR...

...content on subjects such as: CRISPR Chemogenetics Optogenetics Genetic Code Expansion...Overview of Addgene's educational resources, including eBooks, science guides, videos, and protocols ... content , including protocols, tips on using Addgene’s materials, and career advice. Playlists include...Protocols Careers How-to Blog Read about plasmids in Addgene’s repository , and related molecular biology techniques...Concentrations for Bacterial Selection What is a Plasmid? Genetic Code Strain Information Molecular Cloning Techniques...

...). Efficient site-specific integration of large genes in mammalian cells via continuously evolved recombinases...Over 75,000 pathogenic genetic variants have been identified in humans and cataloged in the ClinVar database...editing tools. We’ll summarize the major ones in Addgene’s repository later in the post. Advantages of prime...by double strand breaks has been widely used to generate precise changes. However, the efficiency of Cas9...Prime editing has a lot of promise for treating genetic diseases. In our summer Hot Plasmids post, we covered...demonstration of prime editing used to rescue a genetic disease phenotype in an animal model, which was...s41587-022-01527-4 Additional resources on the Addgene blog PRIDICT: Predicting Efficiencies of Prime...

...Resources on Addgene.org Addgene’s Viral Services Addgene’s Virus Protocols Addgene’s Chemogenetics Guide ...system.   Figure 2: General overview of chemigenetic tools. A genetically encoded component is delivered...Resources on the Addgene blog Viral Vectors 101: Chemogenetics Viral Vectors 101: Optogenetics Fluorescent... within a cell are genetically encoded voltage indicators (GEVIs) and chemigenetic sensors. These tools...on this later). Chemigenetic sensors take a hybrid approach and utilize both a genetic component as well... system. Single Fluor Chemigenetic Sensors This system consists of a genetically expressed enzyme, typically...about how viruses can be used for chemigenetics, biosensors, and generally as biological tools, our blog ...

...AAV1 Chemogenetics Scott Sternson New viral prep pAAV-CaMKIIa-hM3D(Gq)-mCherry AAV1 Chemogenetics Bryan...mCherry AAV1 Chemogenetics Bryan Roth New serotype pAAV-hSyn-hM3D(Gq)-mCherry AAV1 Chemogenetics Bryan Roth... Brian O'Neill.  More resources on the Addgene blog Addgene's Data Hub What is a Virus (and a Viral Vector...but browsing the 49 new preps available in the Addgene viral vector repository with a nice cup of hot ...Table 1: New ready-to-use viral vectors in the Addgene repository  Name Serotype Category Depositor... serotype pAAV-hSyn-hM4D(Gi)-mCherry AAV1 Chemogenetics Bryan Roth New serotype pAAV-EF1α-F-FLEX-mNaChBac-T2A-tdTomato...prep pAAV_hSyn1-SIO-stGtACR2-FusionRed AAV5 Optogenetics Ofer Yizhar New serotype pAAV-CaMKIIa-ChRmine-oScarlet-KV...

...learn what Addgene can do to help. More resources on the Addgene blog Twenty Years of Addgene Sharing: ...researchers around the globe. Addgene’s educational resources As Addgene built out our scientific support...other videos on Addgene’s YouTube channel.   Figure 4: Global viewership of the Addgene YouTube channel...about virus, I learned from Addgene,” or “I learned to clone from the Addgene website.” We delight in seeing...When Addgene opened in 2004, we focused on creating a repository that made it easy to share plasmids,...accessible, due to a wide range of logistical barriers. Addgene worked to identify, reduce, and remove these logistical...accessible weren’t always the same thing, shaped both Addgene’s founding and our growth into the company we are...

...Community Addgene: In 2012 your lab first deposited plasmids with Addgene. In 2014, you joined Addgene's Advisory...lab. In June 2014, Dr. Fujii joined Addgene's Advisory Board.  Addgene: Your lab has worked extensively ... enChIP through Addgene. Retroviral Expression System for enChIP using CRISPR Addgene: Your lab recently...the plasmids of these systems to Addgene in the near future. Addgene: Has your lab been working on any...first knew Addgene around 2010 and found the scheme very nice at first sight in that Addgene takes care...ourselves without Addgene. So, when I received an invitation to join the Advisory Board of Addgene, I accepted...Hodaka Fujii lab's Addgene page and click the "Subscribe to Alerts" link in the Addgene Alerts box. References...

...expected to grow with the ever-expanding genetic toolbox generated by the continuous effort of numerous scientists...). GFP is the most popular, most widely used genetically encoded fluorescent probe. Several factors contribute...probe, it is the first choice when selecting a genetically encoded fluorescent tag. There are, however, ...1998). Another mutation (A206K) is necessary to generate the strictly monomeric EGFP variant, mEGFP (Zacharias..., followed by an intramolecular reaction that generates the chromophore (Tsien, 1998). Importantly, this...Thorn, 2017). These fluorescent analogues are generally added to the bathing solution and require uptake...adding GFP may alter diffusion rates. Smaller genetically encoded probes are based on cofactor binding ...

...If you’ve ordered a pooled library from Addgene, you may have been a little surprised at how this powerful...deposited many types of pooled libraries here at Addgene, including cDNA, barcoding, screening, and gRNA...gRNA libraries. Most of the pooled libraries at Addgene are lentiviral pooled libraries, though other types...types exist.  When you buy a pooled library from Addgene, you can either order a DNA pooled library, which...single tube, or a pre-made viral prep library. Addgene’s pooled libraries are typically DNA preps, but ... A) Amplification, validation, and lentiviral generation of a pooled library. An aliquot of the pooled...experiments. Pooled library lentiviral preps are generally aliquoted to reduce the number of freeze-thaw ...

... institution’s TTO or legal office first. Generally, Addgene is also waiting to receive that legal agreement...materials from Addgene can qualify. We also reward improvement against one’s own record Mention Addgene Master...question? Refer them to Addgene’s Tech Transfer Team: techtransfer@addgene.org Congratulations to the...between a TTO and Addgene, whose terms apply to all future deposits of materials with Addgene Master Reception...between a TTO and Addgene, whose terms apply to all future materials requests from Addgene MTA: material ...commercialization Additional resources on Addgene.org Addgene Help Center: Tech Transfer Technology Transfer...arrival of the newest plasmid you requested from Addgene...the plasmid that will help you run that last ...

... Resources on Addgene.org Learn how Addgene engages with the community on our Addgene gives back page...Initiatives for the Personal Genetics Education Project (pgEd). Advances in genetic technologies and other ...broadens the conversation about genetics pgEd, based at the Department of Genetics of Harvard Medical School...existing inequalities and injustices. The Personal Genetics Education Project (pgEd), where I currently work...work, strives to broaden the conversations about genetics and facilitate dialogue between scientists and...the personal, social and ethical implications of genetics. In these conversations, pgEd strives to be inclusive...and empowered to make their own decisions about genetics, both as individual consumers of technologies ...

...Resources at Addgene.org Find pooled libraries at Addgene Find basic lab protocols at Addgene ...As a tech at Addgene, part of my job, in addition to preparing DNA and generating/replenishing kits, ...information from the workshop. Future workshops for Addgene Addgene strives to be a great resource for the research...Technician at Addgene who is interested in medicine and music.       Additional Resources on the Addgene Blog ... use what we learn to improve processes in the Addgene lab. In this post, I’ll tell you a little bit about...excited to go there because both LabCentral and Addgene aim to make scientific advancement easier. NEBNext...extract libraries from previously fragmented DNA.The general process for library preparation using this kit ...

...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...

...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...

... 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...

...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...

...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...

...', 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...

...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-...

...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...