...  and that’s just to name a few. But what if you want to study RNA-protein interactions? The characterization...RNAs and proteins with UV light, followed by an antibody pulldown of a protein of interest, much like using...disease. RaPID can also be used to test how RNA variants associated with human disease affect protein binding...

...where you want it to? Knowing that a gene is expressed in a particular cell type is important not only ...Gross-Scherf, B., Morikawa, R.K., Rompani, S.B., Hantz, P., Szikra, T., Esposti, F., Cowan, C.S., Bharioke...

... deliver Cas9 variants, such as a transcriptional activator dCas9-VPR, if these variants are fused to ...Mature Nanoblades then bud into the cell culture supernatant and can be purified by centrifugation. Gag-Cas9...

...scientific conference or a karaoke bar), you probably want to sigh. The problem is that many of these studies...Sapozhnikov and Szyf 2021, Charlton et al. 2020). Most importantly, TET proteins affect transcription independent... directly inside the gRNA sequence is a great guarantee that DNMT1 will be blocked (within PAM works, ...

...active in vitro and in human and plant cells. Its small size offers advantages for cellular delivery. Get ...integration and connectivity without the constraints of antibody labeling. This opens the door to in vivo experiments...

...publication.   3’UTRome The 3’UTRs of mRNA are important for post-transcriptional regulation in C. elegans...Dynamics Database (WDDD) The WDDD aims to acquire quantitative data on all the cell divisions of the C. elegans...lab in 2015 adapted the AID system discovered in plants to conditionally deplete proteins of interest in...

...riboprobe binding (via hybridization) specific antibodies that detect the mRNA-riboprobe complex are added...stain the cells that contain this mRNA-riboprobe-antibody complex and, as a result, these cells turn blue...Addgene. If you use these published protocols and want to share pictures, feedback etc. please write to...

... Zhang’s lab characterized two Cpf1 nucleases, distant cousins of well-known Cas9. Cpf1 cleaves DNA in...2015). PubMed PMID: 26422227.   Potential advantages of Cpf1 Type II CRISPR systems based on Cas9 ...efficiency across CRISPR nucleases and NgAgo is an important next step for the field. Based on what we know...

...with all these CRISPR plasmids (and libraries)? We wanted to know, so we classified CRISPR plasmids containing...other countries, from Japan to Australia, are significant depositors and requestors. CRISPR tools are developed...our CRISPR depositors and requestors. Thank you!  Want to dive deeper into Addgene’s CRISPR data? Read ...

...poster you want to know more about? Interested in doing a postdoc in a presenter’s lab? Want to collaborate...breaths the day-of, you’ll handle them with ease. Anticipate questions: When practicing with your colleagues...

...difficult” GC-rich regions of the genome, which seem resistant to Cas9 cleavage (1). Both the CRISPR-Cas9 and...researchers were unhappy with the new method and 65% wanted to wait until someone else optimizes an NgAgo protocol...wouldn’t be a surprise if someone came up with a mutant NgAgo (or a novel endonuclease) protein that can...

...Structure of a retrovirus. Image re-used from Dong & Kantor, 2021 under CC-BY license. Figure 2 gives a...is present in the capsid, as the engineered, recombinant genome is (thankfully) gutted of many essential...1406488. PubMed Central PMCID: PMC372876 Dong, W., & Kantor, B. (2021). Lentiviral Vectors for Delivery of ...

...(ESCs). Your ChIP-seq finds just over 13,000 significant binding peaks for NANOG in the human genome. .... But not all of those 13,000 regions are important for maintaining ESCs. So which of these 13,000 regions...cells compact DNA in a way that brings linearly distant regions close together. gRNA Selection and Filtering...

...After sequencing and searching for cellular RNAs important for Cpf1 function, they found that mature crRNAs...staggered pattern similar to FnCpf1. Potential advantages of Cpf1 over Cas9 Type II CRISPR systems based... Direct synthesis of these crRNAs should be significantly cheaper than that of the ~100 nt crRNA/tracrRNA...

...continue to rely on printed journal articles as the dominant tool for sharing ideas. This leads to everyday...platforms (such as social networking, visual appeal, instant sharing across multiple other sites), I created...learn about the results of studies on reducing antibiotic prescriptions for respiratory tract infections...

... the Addgene repository can save researchers significant time and removes a variable from the lengthy ...from 2016–2023 by type of tool. Equally as important to what is being delivered is where it’s being...trends in our own repository data, giving us significant insight into which vectors are likely to be of...

...fluorescent protein control. We also added the toxin-antitoxin stability cassette Axe-Txe so that the plasmid...plasmid could be maintained in the absence of antibiotics, resulting in the plasmid pBAD-bARGSer-AxeTxe (Fig...agents to boost expression to appreciable levels. We wanted to create second-generation mARGs with higher expression...

...biochemical assays followed by in vivo testing. The mantra being “you can’t trust those whacky cell lines”...contamination with HeLa being the most frequent contaminant, closely followed by T24 bladder cancer cells...repository like the ATCC, there are times when it is important to check their identity. For example, perhaps ...

...fragments are likely non-functional, they could be dominant-negative and/or toxic. A potential solution is..., was 81% of that from a single vector, but significantly greater than the expression from an overlapping...While the hybrid dual vector system has a slight advantage over the overlapping and trans-splicing approaches...

...photo-stimulated through fiberoptic cannulas directly implanted in the injection site and connected to a laser...Taylor et al., 2016; Hooper and Maguire, 2016). Recombinant adeno-associated viruses (rAAVs) have become ... The resulting fusion gene is inserted in the antisense orientation with respect to the promoter to prevent...

...your data by Nov. 12 to compete. The top ten participants will be invited to compete in our speed round...winner each receiving a $500 travel award. All participants will receive an Addgene t-shirt.  Our judges...

... suggestions in the comments.  X has moved significantly away from the functionality and features that...this reason, our timeline has accelerated. We significantly reduced posting on November 20, 2023, and as...

...more commonly known as fruit flies, have been an important animal model for research, particularly genetics...well, annoy. Fly-related Issues in the Lab A significant drawback of working with fruit flies is their...

...tube-labeling, quick plate pouring, and much more. We didn’t want to keep the knowledge we’ve gained along the way...trick When combining reagents in the lab, it is important to mix them thoroughly before use.  For efficient...

...worth of ladder. The Penn State ladders are significantly cheaper. When using a homemade plasmid prep ...increases the costs to $20-30 per ladder: still significantly cheaper than commercial ladders. Are you interested...

...story of Rosalind Franklin and her incredibly important role in solving the structure of DNA. 2. The...her or her family's knowledge. 3. Fossil Rock Anthem One of Tom's earliest and most popular videos. ...

...ventilation, and lab lighting/HVAC requires a significant energy expenditure here at Addgene. These are... critical systems that cannot be removed or substantially reduced in an attempt to save energy. The transition...

...biologist Tips for protein purification Replica plating Anti-CRISPR Retroviral vectors Career development for...for the Addgene blog simply email us. We can’t guarantee that all those who sign up will be able to write...

... focusing on safety and the ability to generate mutant microbial strains faster than ever before. 29:54... end : How Alex hopes to help Addgene advance - plants for trimmed down protocols, distribution of new...

...References 1. Choi, Vivian W., et al. "Production of Recombinant Adeno‐Associated Viral Vectors for In Vitro and...18265393. Additional Resources on the Addgene Blog Important Considerations When Using AAVs AAVs Are Great ...

...that you can stage in a lab diorama. Or maybe you want to create a stack of wrapped gifts using life science... festivities - however you celebrate! And if you want to see how Addgene celebrates and decorates, follow...

...in CRISPR including: Base editing Prime editing Anti-CRISPRs … you name it! So get excited and download...multiple genes in a cell population. The resulting mutant cells are screened for phenotypes of interest. ...

...search bar Now, you can search for the articles you want to find without even leaving the Addgene blog! Just...articles on the Addgene blog by subscribing to instant alerts below.  ...

...toiling away in laboratories this holiday season want the work you are doing to have an impact, to move...libraries and offering another way to consider and quantify the impact of scientific publications. Ventures...

...expression level without the need for a custom antibody. There is one major caveat - a tag may interfere...affinity for other proteins, and accessibility to an antibody. From their initial pool of candidates, they selected...

...week. So I have developed some good habits that I wanted to pass on to you to make sure you are getting ... you properly visualize your results.The most important of those is to always look closely at the trace...

...water of HR work. What do you think is the most important resource that Addgene provides the scientific ... how to operate during the pandemic, but it’s important to develop a plan for moving forward. Sooner or...

...bioluminescent, some types of corals and anemones are vibrantly colored. He hypothesized that GFP-like proteins...they’re used, see some new techniques that take advantage of them, and give some tips on deciding which ...

...accomplishments. Employers and academic departments want to be able to tell that you know about something...Learned Sooner by Peggy Klaus Looking for a mentor or want to start a peer mentoring group? "What Makes a ...

...choosing a specific serotype, but occasionally you may want to further control where your gene of interest is... many more tips and ideas on how to use these fantastic tools in your experiments.  Many thanks to Hongkui...

... they’ve engineered SECURE adenine base editor variants using a protein-truncation strategy with reduced...Transient expression of CRISPR/Cas9 in cells has advantages like reduced off-target activity and decreased...

...and see if we should start celebrating papers and grants too with pizza or pub.” The Cepko Lab celebrates...required to achieve success in the lab, it is important to take the occasional breather and acknowledge...

...Organize you experiments, keep track of upcoming grant deadlines, and sync everything to your calendar.... updates to protocols in real time. You may also want to consider moving your paper spreadsheets for reagent...

...double strand breaks. Researchers have long taken advantage of this natural process to integrate protein tags...selectable gene product for isolation of successful transformants. Addgene distributes several ready-to-use, modular...

...increasing transgene cargo capacity, enhancing anterograde transport efficiency, and engineering viral capsids...development of this practical guide by the workshop participants  - a testament to openness and sharing in science...

...physical scientists are underrepresented. It’s important to note that these results could be biased for...Resources Check out the Top 50 Science Stars on Twitter Want suggestions on how to use social media as a scientist...

... for you to find the plasmids and publications relevant to your research interests. We do this by promoting...other resources. If you think we’re missing an important collection or if you think a particular plasmid...

... Society, or maybe you think about a local food pantry or community arts organization. Many people are...resources continuously raising money from donations or grants to fulfill their mission. Addgene is fortunate ...

...Experiments Experiment  Detection Method Notes Quantification of a process Plate-based assays (plate reader... HPLC (proteins or other molecules) Isolate or quantify cells, proteins or other molecules that have been...

...worthless. But simply writing down your data doesn’t guarantee a publication. There is an art to composing manuscripts.... Practice writing everyday Overall, the most important instruction the classes taught me was to write...