We narrowed to 1,054 results for: STI

...sure it is working? This article addresses these questions, which are critical in setting up a FRET-based... the wrong address in a different city, you can still develop a successful FRET-based assay since energy...interest: you just don’t know it yet! The poly-histidine tag often used for protein purification is a reliable...dye, as well as by extending the length of the histidine tag from 6 to 10 residues. While these Cy/NTA ...incorporation of fluorescent probes into protein: hexahistidine-tag-mediated fluorescent labeling with (Ni2+...FRET-Based Approach for Antibody Detection–A New Serodiagnostic Concept." PloS one 8.5 (2013): e62739. PubMed...

...efficiency of multiplex gene editing. Moreover, existing multiplex strategies have been facing difficulties... fused base editors (nCas9-BE), as Cas9 is mechanistically more favored for efficient base editing. They...human cysteine tRNA stood out from their initial testing. It outperformed other tRNA candidates (either ...multiplex genome engineering of crops or allow for sophisticated CRISPR screenings that would perturb the unprecedented... only minimal genetic payloads. Different from existing multiplex array architectures, including previous...

...disorders. Antibodies administered as drugs are still immunogenic, meaning that they elicit an immune ...previous exposures to that specific antigen, pre-existing conditions, etc. that can vary widely between ...decisions about in vivo application and clinical testing. This would pave the way for improved patient outcomes...Burns-Naas, L. A. (2015). Chapter 27—Immunotoxicology Testing of Monoclonal Antibodies in Macaca fascicularis...Containing Unnatural Amino Acids: A Method Using Existing in silico Algorithm Infrastructure and a Vision...

...Grammarly also offers basic writing suggestions, like alternative word suggestions. De-jargonizer The De-jargonizer... at these graphs when I need that extra push to stick with my writing routine. The graph of which days...detecting the readability of your writing. It makes suggestions on how to make your writing less complex. While...While I don’t think you should accept all of the suggestions Hemmngway makes (it wasn’t designed for technical...

... also be used to engineer new pathways with sophisticated gene expression controls. Most cell types do...two-component systems (TCSs). TCSs consist of a histidine kinase that phosphorylates a response regulator...domain from cyanobacteria to a common E. coli histidine kinase. This hybrid construct senses light, and...light; these two systems can be coexpressed for sophisticated control of gene expression. An E. coli light-sensitive...light-sensitive two-component system (TCS). A hybrid histidine kinase/photosensor senses various light inputs...

...when seeking your dream lab. The principal investigator's point of view If you are looking to join a ...course, the specific research field should be interesting; however, this should (in most cases) not be ...be the driving factor. The following questions can serve as a guide for the quest of a perfect "fit". ...Scientists". This post also includes a useful list of questions for a candidate to ask lab heads, lab members ...human disease. For more details and many more suggestions on planning for future success by finding and...

...or +1 (617)-ADDGENE (233-4363) with additional questions. Bacterial Strain Addgene can distribute your...Return to Article" button to return to the table listing all of the plasmids for this deposit. You can then...deposit, associate the plasmids with a Principal Investigator (PI), provide us with your shipping address ...hereby certify that I am affiliated with the institution ___. If this is no longer true, please modify...material under the terms deemed necessary by my institution or other third party rights holders. Once this...January 2016. It was updated by Angela Holmes and Christina Mork in February 2024.  Additional Inside Addgene...

...Biology. Can you tell me where to start?” is a question more and more students are asking. Teachers are...of educational programs all focused on current questions and experiments in the field of synthetic biology...from her own MIT curriculum. Drawing on current questions being asked in MIT’s research labs, BioBuilder... into modules that students and teachers can investigate together. Teachers decide which BioBuilder lessons...ideas ranged from a biological breathalyzer to pesticide breakdown, production of fire-resistant housing...

...Since there are many thousands of partitions, statistical modelling can be used to determine the actual...mismatch nuclease assays (2, 6, 7). Ability to distinguish between mono-allelic and bi-allelic mutations...digital PCR assays is that they are easily able to distinguish single-cell derived samples with mono-allelic...instructions on how to design such assays, in addition to testing their performance (2). Remember that although digital... digital PCR is a great screening tool, it still requires target DNA sequencing to confirm the exact nature...

...reach out to the program coordinator with your question(s). This precautionary step increases the likelihood...communicates your interest in a specific program, and institution. Consequently it’s best to draft a cover letter...vast majority of programs tend to ask similar questions, thus devoting some time to writing a solid personal...interview. Formulate and practice a set of common questions with a peer or mentor. Several days prior to the...entry-level position following graduation. While these suggestions increase your competitive edge, they alone, are...

... the fragments size of your digestion on an agarose gel. If the digestion does not provide the fragments... to your cloning strategy: Restriction enzyme digestion Be mindful of the restriction enzymes you chose...like XbaI, ClaI, etc. If you are doing multiple digestion be sure that the buffers and temperature are compatible...It's also best to use 1-2 ug of the vector for digestion. PCR Design primers that overlap at least 24 bp...

...antibody region that binds to the antigen) from existing antibodies. Each cleaved antibody will produce...(Muyldermans S., 2013). They have the unique distinction of being the only affinity reagent on this list...  To antibody or not to antibody?  That is the question… or is it? This post isn’t meant to convert anyone...trendy as that may sound. Validated antibodies are still an excellent option for many applications, and frankly...References Grumezescu, A. M. (2018). Drug targeting and stimuli sensitive drug delivery systems. Elsevier Science...

...researcher at Lunenfeld-Tanenbaum Research Institute, posting about Addgene’s 20th anniversary on Twitter...SirHamhands, responding to a request for “any suggestion for CRISPR online course for beginners” on r/...alike. We transformed this process by aligning institutions with standardized agreements and electronic ...!” — Julien Béthune, Professor at HAW Hamburg, posting about his Blue Flame Award on Bluesky. In 2016,...

...cytosine and adenine base editors that can be reconstituted in vivo to form a protein identical in sequence... plasmids and co-transfected into cells for reconstitution. Image from Levy et al., 2020 with permission...when paired with appropriate fluorescent optics. Existing red fluorescent voltage indicators, however, fall...neural circuit manipulations when combined with existing green-fluorescent GEVIs and blue-light-sensitive...The kit functions in the model cyanobacterium Synechocystis sp. PCC6803, and the fast growing strain Synchococcus...

...available to counteract the toxin. If, however, an investigator were to successfully clone an insert into the... (re-ligated empty vectors, for instance) would still express ccdB and therefore would die. This procedure...insertion will completely replace ccdB with the investigator’s insert of interest. Hence correct clones are... behind the “swapping out” of ccdB with the investigator’s gene of interest.   CcdB resistant E. ...further. Feel free to offer your comments and suggestions, and even to submit a guest blog entry of your...

...controlled by deliberate and compulsory actions instigated by the central nervous system signaling to peripheral...metabolic syndrome. Coregulators Coregulators constitute a group of some 400 molecules that have been ...regulated manner. The fact that coregulators belong to distinct complexes may explain how more than 300 different...collectively address many important and fundamental questions in nuclear receptor and coregulator research. ...the NURSA website will feature direct links to listings of available plasmids for our curated NRs and ...

...ball-and-chain that they resemble, actually impede your investigation into the regulation of your favorite gene. But...these studies report correlations without clearly testing whether a certain methylation state causes a condition...to create rare DNA bases that are recognized by distinct proteins and result in unclear transcriptional...achieved using lentiviral backbones expressing blasticidin or puromycin resistance genes.  It’s that simple...Feinberg A, Vogelstein B (1983) Hypomethylation distinguishes genes of some human cancers from their normal...

...penicillin resistance plasmid can grow in spite of a hostile Penicillium fungus trying to kill them off. While...resistance to toxic substances like heavy metals or pesticides, enabling their hosts to survive where others... This ability to endure a harsh environment is distinct from the benefit of resistance plasmids, which...shifted over the years, and new classifications are still being created as whole-genome sequencing becomes...Plasmids Isolated from Lactic Acid Bacteria:Their Characteristics and Potential Applications in Biotechnology...

...how close is close enough for the desired edit to still occur if the cut site is sub-optimal? As with everything... cut site is just 10 bps away from the site in question (Kwart et al, 2017). To add insult to injury, ...same PAM preference of NG was isolated by the Kleinstiver Lab (read more on this lab's other developments...Nonetheless, in an AT-rich region, these Cas9s may still leave you choosing between efficiency and location...location.   Nearly PAM-less editing   The Kleinstiver lab sought to address the PAM problem through structure-guided...

...certain cell types.  Botond Roska’s lab at the Institute of Molecular and Clinical Ophthalmology Basel ...finding retinal cell-type specific promoters. Most existing promoters express in a broad number of cell types...targeting the same cell class in all three species are still greater than random chance. The chances of a promoter...human retinas but not in mice was low (R=0.12), suggesting that mice could be used to eliminate promoters...Morikawa, R.K., Rompani, S.B., Hantz, P., Szikra, T., Esposti, F., Cowan, C.S., Bharioke, A., Patino-Alvarez,...

...a risk that they may underestimate viral titer as these assays cannot distinguish between a cell that ... based off of p24 quantification tend to be overestimates since they can include free p24 and defective...assay, titering via measurement of viral RNA can overestimate the amount of infectious virus in a prep as ...integration events, reducing the possibility of underestimating the viral titer. The main drawback of this ...

...engineered precursor of the systemic capsid in question (or both). Therefore, a high-efficiency systemic...These are only starting recommendations; you will still need to experimentally determine the best titer ...your experiment, it’s time to start selecting and testing systemic capsids! Figure 2 contains a decision ...reduced efficiency, and is not neutralized by pre-existing antibodies against AAV9-based vectors (and vice...AAV-PHP.S, with a bias toward segments of the small intestine (Chen et al., 2022). Systemic capsid for alveolar...Cre-dependent selection yields systemic AAVs for targeting distinct brain cell types. Nature Methods, 17(5), 541–550...

...in 5’ four base overhangs after cutting. The destination vector receives the assembly after similar Type...re-digested and "eligible” for ligation into the destination construct. Reactions are often cycled between...acquisition of antibiotic selectable markers by the destination plasmid. Antibiotic selection allowed for high...very occasional uncut or cut/re-ligated pGGA destination plasmids. A final validation of the 5-, 12- or...is dependent on the presence of every module, destination construct and functioning Type IIS restriction...

...the prep during the production process? These questions are especially important when producing multiple...we have analyzed hundreds of viral vectors and estimate that recombinase-independent recombination occurs...below to let us know if you have any specific questions about viral DNA NGS you’d like us to discuss. ...by phone or at [email protected] to answer your questions about our quality control process! The images...Vector Characterization Methods for Quality Control Testing of Recombinant Adeno-Associated Viruses. In: Methods...

...protocol for your pooled library deposit. When a requesting scientist receives a pooled library from Addgene...colonies, you’ll likely rely on serial dilutions to estimate transformation efficiency (Figure 2).    ... Figure 2: Example of a serial dilution to estimate transformation efficiency. Figure used with permission...protocol paper by the Zhang lab, and the Broad Institute’s general pDNA Library Amplification protocol are...depositing it with Addgene. If you have any specific questions about designing your library amplification protocol...

...discuss the work, set expectations, answer any questions, and (most importantly) decide on a timeline. ...regularly, make yourself available, and encourage questions. Unless you’re delegating a recurring task to ...to a seasoned report, there should be questions! Remember that delegating is not just a way to create more...team members by giving them new opportunities. I still need to check myself sometimes to make sure I am...major problem. But I also learned to ask specific questions. “How’s everything going?” is likely to get you...

...Lance-Byrne and Alex Chavez, researchers at the Wyss Institute for Biologically Inspired Engineering. CRISPR/...protospacer. The resulting double strand break (DSB) stimulates DNA repair mechanisms that can be exploited to...are limitations to a system that relies on two distinct Cas9 variants—nuclease-competent or nuclease-null—in...bacterial species, each of which interacts with a distinct gRNA that allows users to decide which Cas9 protein.... When complexed with a truncated gRNA, Cas9 is still able to bind DNA but is no longer competent to cut...

...other split systems where light is required to reconstitute enzymatic activity. Photoactivatable genome ...(Kawano et al., 2015). Nicknamed paCas9-1 and consisting of the fusion proteins N713-pMag and nMagHigh1...far-red light-activated split-Cas9 (FAST) system constitutively expresses the C-terminal fragment of Cas9 while...Michael Lin, who developed a chimeric protein consisting of Cas9 and a photo-dissociable dimeric green... prevent DNA binding in the dark. Upon light stimulation, the pdDronpa moieties move apart, allowing DNA...LOV2 has a C-terminal helix that unfolds when stimulated by blue light. When fused to the catalytic site...

...student at Stanford University. Arabidopsis is a fantastic model organism for many reasons, not the least...Agrobacterium tDNA insertion likely precipitated domestication of sweet potatoes (Kyndt et al., 2015)! It sounds...too old to transform. This transformation could still work, but the efficiency will be low. Tips: Use...plants: If you transform early, your plants will still have plenty of time to produce seeds after you expose... plant’s flowers inside. 50 mL falcon tubes or plastic weigh boats are both great options for this, depending...

...inactivation of the vector can still occur even in the case of no previous immunity existing for that adenoviral ...challenges remain (Wold & Toth, 2013).  While the pre-existing toolbox for adenoviral vectors in Addgene’s repository... – and we hope that it inspires further use of existing vectors, as well as research into the development...M. (1999). Ovine Adenovirus Vectors Overcome Preexisting Humoral Immunity against Human Adenoviruses In...

...phenotypes. Two labs, the Deverman lab at the Broad Institute and the Wilson lab at the University of Pennsylvania...the brains of C57BL/6J mice and BALB/cJ mice, suggesting it was a likely player in PHP.B permissiveness...receptor (AAVR). Cell lines lacking galactose were still transduced by PHP.B but only if Ly6a was expressed...with PHP.eB, an enhanced version of PHP.B, virus still reached the brain endothelial cells of these mice... available from Addgene. References Holmes, Christina, and William L. Stanford. "Concise review: stem...

...synthesis (those of us who have used antibodies for diagnostics know how frustrating it can be to get a “bad ...the bound protein fluorophores and quenchers are still relatively bulky. One of the advantages of oligonucleotides...stable and brighter version of the aptamer, but it still had limitations in vivo. Using a combination of ... a lower magnesium dependence. Though initially still unstable in eukaryotes, the Jaffrey lab has since...et al. “Aptamer-based assay for prion diseases diagnostic”. Procedia Environmental Sciences. Volume 12,...

...full length gene and/or protein is reconstituted. One way to reconstitute a split protein like Cas9 is to...allow for more room for regulatory sequences while still fitting Cas9 and its gRNAs into one AAV, Cas9 must...confirming its suitability for in vivo studies. Testing AAV-SaCas9 in vivo To test the efficiency of AAV-SaCas9...Nogales E, Doudna JA (2019) CasX enzymes comprise a distinct family of RNA-guided genome editors. Nature 566...

... 1–9 (2017).Walton, R. T., Christie, K. A., Whittaker, M. N. & Kleinstiver, B. P. Unconstrained genome... effects. The SpG variant, discovered by the Kleinstiver group has a PAM of NGN (Walton et al., 2020)....Don’t want to permanently modify the genome but still want to affect gene expression? dCas9 can be used... and antibody-activator fusion proteins. The Synergistic Activation Mediator complex, generally the strongest...

..., black pellet was sent to the AbuGoot lab for testing, they weren’t sure what they would find… but it...new CRISPR effector protein they named Cas7-11. Testing showed that Cas7-11 cut RNA with unprecedented ...insertions, and deletions. But CRISPR’s uses were still limited to small deletions and small insertions,...that gap. Consider, for instance, a disease like cystic fibrosis, which can be caused by thousands of different... Yarnall, Cian Schmitt-Ulms, Rohan N. Krajeski, Justin Lim, Lukas Villiger, Wenyuan Zhou, Kaiyi Jiang,...

...range of logistical barriers. Addgene worked to identify, reduce, and remove these logistical barriers...for people depositing plasmids and for people requesting them. Prior to this, many plasmids were available...packages navigated customs and arrived at their destination in a timely manner and at the correct temperature...to scientists across the globe?” By asking this question, and by listening to the answers from our community...

... during the early stages of the process. Some questions to start with are: Is your sequence of interest...opposite (reverse) orientation? The answers to questions like these will determine which acceptor vectors...assembly efficiency. This can be done through domestication, which is the modification of a DNA sequence... the synonymous mutation of a codon. You can domesticate the sites yourself using PCR, but gene synthesis...gene synthesis, you can request the sites to be domesticated through the process, and request your sequence...

... plasmid is delivered to your target cells, you still need to select for the cells that contain it. Antibiotic... approach in a mammalian cell experiment should still include an antibiotic resistance gene for growth...express your protein of interest. However, you do still need a species-compatible promoter for expressing... consider the viral vectors themselves to be a distinct gene delivery method. We focused this post on ...between prokaryotes and eukaryotes. Conclusion I still think it’s pretty incredible that my cells “speak...

...was applying. I probably did not respond to her question as well then as I would now. I would have to repress... repress laughter if I was asked that question today, because I sometimes wish for a stretch of time that...me, “I find that at a desk, it is much easier to stick to timelines. This is probably because you don't...careers and desk jobs “alternative careers.” Realistically, based on the numbers of PhDs and available ...into science in the first place. Finally, if you still find the idea of not holding a pipette again upsetting...

...the way we apprehend molecular complexes and is still a very useful tool.    Like a guitar hero (that ...SYFP2A, mTurquoise/mVenus and others. CFP-YFP is still one of the best and most used pairs to measure FRET... is difficult to address especially when you investigate unknown molecular complexes. But it is a parameter... faster measurement systems, this technique has still many good days and good “gigs” ahead...No offense...Chemiluminescence Resonance Energy Transfer    Interesting Articles About FRET: Piston DW, Kremers GJ. ...