...AAV serotypes, and the ever-growing variety of recombinase systems. Picking Ligands Picking the right ligand...ligand co-developed by Bryan Roth's and Takafumi Minamimoto's lab called deschloroclozapine (DCZ) shows promise...our chemogenetics poster! This blog post was originally written by Maya Peters Kostman in May 2020 and...

... incident, and the sterilization of women in marginalized communities have created a legacy of distrust...included taking extra coursework and attending seminars in bioethics, public health, science policy, and...advantage of their academic environment and attend seminars, symposia, or courses in social, health or policy-related...

...is ideal because it eliminates the potential of human error or cross contamination between samples. With... Shannon Rinaldi...

...immunogenic profile. AAV was first discovered as a contaminant of adenovirus preparations. It is an icosahedral...untreatable until 2008. Following a single dose of subretinal administration of rAAV2 carrying the human gene... responses to adeno-associated virus and its recombinant vectors." Gene therapy 10.11 (2003): 964-976....

... the protein or structure of interest. For a C-terminal tag, as shown in the example above, we inserted...would be transcribed and translated. For an N-terminally tagged protein, we used the same strategy, inserting...tagging using CRISPR/Cas9 in human stem cells to illuminate cell organization." bioRxiv (2017): 123042. Additional...

...majority of our plasmids and pooled libraries were originally described in a peer-reviewed scientific publication... our thousands of depositors. That means their original articles are your best bet to figure out how to...asking to avoid unrealistic expectations. And finally, provide feedback. If you are unsatisfied with ...

...dCas9. Within a few rounds of cell division, the original methylated strand is diluted to undetectable amounts...dCas9 so that it may later be excised by Cre recombinase. Addgene has several great options: pLV hUbC-...L, Li C, et al. (2018) OGT binds a conserved C-terminal domain of TET1 to regulate TET1 activity and function...

...Originally published Apr 30, 2020 and last updated Jul 12, 2021. What started out as a small collection...used for this study at Addgene which include a C-terminal deletion in the spike protein to improve expression...pseudoviruses to determine how effective sera from vaccinated individuals are at neutralizing the different...

...PCR product that comes from amplification of the original plasmid which can over-inflate results. A packing...vectors carrying fluorescent markers are used as it eliminates the need for antibody staining. Check Out Our...into a target cell’s genome. This approach can discriminate multiple integration events, reducing the possibility...

...researchers to use. Although we haven’t yet made our final target decisions, we intend to focus on motor neuron... a new project at a makerspace is finding the financial resources to pursue it. Since makerspaces are ...equipment for new uses. We’ll probably never use the Illumina GAIIx we recently acquired  for sequencing again...

...This post was originally published on LinkedIn. Follow Addgene on LinkedIn for repository news and updates...members all say the same thing and one panel member dominates the discussion? Or maybe the one that goes way...front of a crowd. The moderator will then develop a final list of questions for the actual event, based on...

...Figure 1). Through this method, we found key determinants of prime editing efficiency such as the length....0 while accounting for some of the inherent inaccuracies of computational prediction models. In addition...efficiencies using sequence features and DNA repair determinants. Nature Biotechnology 2023, 1–11. https://doi.org...

...2014 to 2023. As you can see, nucleases, the original CRISPR tool, remain the most popular deposited...Read our recent publication, “The Expanding Dissemination and Distribution Patterns of Diverse CRISPR ..., R., & Barrangou, R. (2023). The Expanding Dissemination and Distribution Patterns of Diverse CRISPR ...

...generated info for your own purposes, or delete the originals if desired. It works as a whole or in part. For...ranging between 0 and 1. The cutting frequency determination (CFD score) is used to measure specificity of...re-binding. Lastly, the sequence inserted by HDR may inadvertently re-create a match for the active guide. With...

...don’t get done if there is not a clear deadline. Finally check-in regularly, make yourself available, and...that one on one discussion can be confidential. Finally, provide timely feedback. We often associate feedback...manner. If someone’s performance or behavior is inadequate, you need to address it as soon as possible for...

...increased sensitivity allows for a more confident determination of copy number, albeit within a range of RQ ...trends? How confident do you need to be in your determination to move forward with your experiment?  Complexity...real-time polymerase chain reaction methods for the determination of amplification of Plasmodium falciparum multidrug...

...the end product of PCR temperature cycling. The final PCR product is often visualized on a diagnostic ... dNTPs and other reagents to run low during the final cycles which would slow or stop PCR amplification...statistics to determine the concentration of the original template DNA in the sample.  Addgene currently...

...active during M and early S phases, when HDR is inactive, and the balance of NHEJ, HDR, and MMEJ repair...longer the homology, the more efficient the recombination, but with longer homology arms comes more time...which has been difficult to modify due to NHEJ’s dominance over HDR in this species. As CRISPR technology...

...for some applications, the benefit it offers is marginal. Fortunately, amp can easily be replaced with ...NPTII, facilitates phosphorylation of kan, which inactivates it. The NPTII gene confers resistance to several.../1475-2859-11-56 Trastoy, M. O., Defais, M., & Larminat, F. (2005). Resistance to the antibiotic Zeocin...

...labs and institutions. For example, search for “Melina Fan” and you’ll be able to go directly to a page...In this example we’re going to be looking for recombinant monoclonal antibody plasmids from the James Trimmer... ‘Vector Types’ I selected ‘Lentiviral’ which eliminates most of the plasmids designed for other uses....

...are binary gateway vectors and can be used for promoter swapping and generating N and C terminally tagged...4. Nakagawa, T. et al. (2007) ‘Improved Gateway Binary Vectors: High-Performance Vectors for Creation ...

...template control PCR reaction for detecting contamination is also a good idea.    Analyzing PCR product...positive clones (+) compared to negative clones. Finally, orientation-specific primers (3) give the same...Resources Promega Overview for Screening for Recombinants New England Biolabs Overview of Colony PCR ...

...virulent or antibiotic resistant bacteria, and inappropriate antibiotic usage selects for resistant bacteria...including Chipotle and Panera Bread have pledged to eliminate the use of antibiotics by their suppliers, and... DSB occurs on a plasmid, the plasmid will be eliminated from the bacterium, which may also induce cell...

...scientist. When our elementary school needed a new coordinator for the annual Math & Science Night, I took the...It’s through the PTA that I recently started coordinating the annual Math & Science Night, a chance for...contacting our school district’s science curriculum coordinator with minimal luck, but eventually the 8th grade...

...fragment to the N- and C-terminal end of each PIP and empirically test which combination provides the best fluorescent...cellular functions, including ubiquitinoylation, kinase signaling, integrin signaling, and transcription...

...T. R. (2015). Stability and compatibility of recombinant adeno-associated virus under conditions commonly...Methods Matter: Standard Production Platforms for Recombinant AAV Produce Chemically and Functionally Distinct...Identification of factors that contribute to recombinant AAV2 particle aggregation and methods to prevent...

...signal sequence (gene start) and a transcription termination/polyadenylation signal sequence (gene end). A...encoded in separate (and thus smaller) plasmids. Finally, having four separate plasmids allows researchers...it reaches the gene end signal, where it will terminate transcription and polyadenylate the mRNA. These...

...isoforms of these proteins, as well as the viral long terminal repeats (LTRs) are different. As a result, lentiviral...modified the 5’ LTR of the transfer plasmid to eliminate the need for the HIV Tat protein and consequently...initial infection. SIN is shorthand for self-inactivating, which is achieved by deleting a large portion...

...Addgene, you’ll usually receive an aliquot from the original library constructed by the depositing lab. (For... library should be very similar to that of the original library. Like amplification, this step also depends...concentration.  Screening time Phew! You are now (finally!) ready to use your transduced cells to perform...

...relating to reproducing the original research can be documented -- in the original context in which it was...import Word documents into RSpace, maintaining the original formatting. In this way data is moved from a static...