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...similarly useful across the diverse kinds of proteins that one finds in cells. High yield of reaction ...freedom to test out lots of combinations, to end up finding something surprisingly useful. To apply these principles...HaloTag modules, which can connect proteins to any kind of small molecule probes with different functionalities...Exchangeable HaloTag Ligands HaloTag Reversible binding of fluorescent ligands to HaloTag PCV-Cas9, Cas9.... Applications are quickly expanding with these kinds of Click Biology reactions. For basic research, ...proteins. Split inteins are particularly useful for a kind of cellular computation, making possible multiple...

... Resources on Addgene.org Find Empty Backbones with Fluorescent Tags Find Plasmids for Subcellular Localization...are visible. These whorls can be quantified for individual cells, and the number of 'normal' looking cells...the results of the OSER assay, you should keep in mind that the presence of whorls is probably concentration...monomeric behaviour in cells (Shaner et al, 2013; Bindels et al, 2017). Still, the data presented in papers...is detected by Constantini et al (2015) and by Bindels et al (2017) but not by Cranfill et al (2016))....build a valuable resource with tools and data that indicate the performance of (a set of) fluorescent proteins...

...days prior to the implementation date, our data indicated that, while 69% of all postdocs were expected ...stakeholders. However, while most institutions are indeed going ahead with their planned raises, some are...had that increase cancelled. Rounding out the remainder, 2% of postdocs who weren’t going to get a salary... reversed their position and decided to allow individual departments to go ahead with the raises as they...detrimental effect on the morale of postdocs, and indeed we have been contacted by a number of postdocs ...to reach out to institutions with a statement indicating that we encourage them to keep raising salaries...Variety of Science Careers Resources on Addgene.org Find Career Opportunities at Addgene  ...

...Background with Our CRISPR Guide Pages Find CRISPR Plasmids Find More Tools for Combinatorial Engineering... Wong. The complexity of biological systems can hinder our attempts to study and engineer them, but what...Gilbert et al., 2014; Konermann et al., 2014) individual genes for studies interrogating their functions...incredibly useful for the efficient generation of individual (and multiple) genetic perturbations in single...sequences via oligo synthesis using the format indicated below and pool-clone them into the pAWp28 storage...identifies genes essential for West-Nile-virus-induced cell death." Cell reports 12.4 (2015): 673-683....

...a look in a typical lab freezer, you’ll probably find lots of different plasmids containing the same protein...Leu28Cys mutation in eDHFR does lead to covalent binding of the TMP-tag ligand (Gallagher et al., 2009)....based on a bacterial haloalkane dehalogenase that binds a synthetic chloralkane (CA) ligand. The current...before investing in one approach or the other.   Finding the right tag for you Protein-based tags typically...Promega (though if you are comfortable with it or can find a colleague to help you, synthesizing your own ligands... experiment, and explore Addgene's collection to find self-labeling fusion proteins or cloning backbones...An In Vivo Covalent TMP-Tag Based on Proximity-Induced Reactivity. ACS Chemical Biology, 4(7), 547–556...

... alternative evidence and discard inconvenient findings. Yet, we cannot prosper in a world where our medicines...ominous political climate in which American scientists find themselves today, and where Canadian scientists ...the public conversation and to tell the stories behind our research.  That's why I marched for science...University of Michigan campus speakers thoughtfully reminded us to use the same critical lens on our scientific... Aguilar Galileo wrote, “now you would not hide behind these two fingers the ninety-nine cubits of Aristotle...promote harmful policies that benefit polluting industries.In March, three climate scientists and an economist...specific, technical manner, so much so that we often find it difficult to convey the excitement and importance...

... on Addgene.org Find CRISPR Plasmids for Your Research Interested in light-inducible Cas9 activity? Check..., 2018). For example, AcrIIA4 mentioned above indicates that it’s the fourth Acr protein discovered that...CRISPR activity one of three ways: Inhibit DNA binding Example:  AcrIIA4 blocks Cas9’s interaction with...2019). Inhibiting DNA cleavage Example: AcrIIC1 binds to the HNH endonuclease domain of Cas9 and prevents...such as inhibiting crRNA loading, inhibiting DNA binding, or inhibiting DNA cleavage.   Five ways to ...of CRISPR activity Regulating Acr proteins by inducible promoters, light, or small molecules could achieve...

...interest  Figure 1. When ABEs are in action they bind DNA so that the TadA domain can edit the short single-stranded...creation of accidental insertions and deletions (indel) by the catalytically impaired D10A nickase from... efficiency in a human cell line while reducing indel frequency more than 90% compared to ABE7.10. Choosing...changes in cellular DNA and RNA dependent or independent of the gRNA used (Gaudelli et al., 2017). The...in this work did not cause genome-wide, guide-independent off-target deamination, an important attribute...0491-6 Additional resources on the Addgene blog Find more blog posts about base editing Learn about another...Addgene.org Get the basics of CRISPR in our CRISPR guide Find all plasmids for base editing in our collection ...

...vector blog posts Resources at Addgene.org Find AAV plasmids Find ready-to-use AAV viral preps Browse all...do this, the group developed a new strategy for finding retinal cell-type specific promoters. Most existing... mouse cell type specific transcription factor binding sites interspaced with random sequences.   Strategy...transcription start site. TFBS: transcription factor binding site. Image from Jüttner et al., 2019 with permission...that was active in the outer part of the retina.  Find the AAV synthetic promoter plasmids! Comparing synthetic...Read more about using AAV for retinal gene therapy Find more about precise neuronal targeting using AAV ...

...of upfront research you have to do to find said robot and to find out how you can get one. As a starting...at Addgene so we’ve been putting some work into finding robots that’ll take a little labor off our hands... pod can pick up 8 tips and position the tips indepently making it easy to spread out your samples. This...live bacteria. For these shipments, we’ve needed to find a way to purify DNA from many bacterial samples ...same time as opposed to preparing each sample individually. Enter the BIOROBOT! This robot is called the...Barcodes to Track Samples Resources at Addgene.org Find protocols for standard molecular biology techniques...

...dividing cells. Find tips on using gamma-retroviral vectors in our Gamma-retrovirus guide and find popular packaging...tetracycline inducible, 3rd generation lentiviral vector backbone from the Shih lab. You can find more lentiviral... vectors. These vectors can induce combinatorial constitutive or inducible gene overexpression, knockdown...-lox system is a technology that can be used to induce site-specific recombination events and is widely...

...trials involve identifying a therapeutic gene, finding a means to deliver it and identifying a suitable...significant clinical efficacy against a form of blindness known as Leber’s congenital amurosis (LCA). This...RPE65. It is characterized by childhood onset of blindness and was thought to be untreatable until 2008. ...their efficacy:  Pre-existing immunity: Many individuals already have wt AAV in their bodies and may therefore...interfere with virus-cell interactions and seriously hinder gene therapy outcomes. Anti-AAV antibodies developed...decline in vision with gene therapy in childhood blindness." New England Journal of Medicine 372.20 (2015...Quality Control Process Resources on Addgene.org Find ready-to-use AAV in Addgene's Viral Service Check...

...this protein is well suited for genome editing.  Find Cpf1 Plasmids at Addgene   Image attribution...red) or LbCpf1 (blue). Mismatch positions are indicated below each graph. Figure adapted from Kleinstiver... and LbCpf1 ribonucleoproteins (RNPs) failed to induce off-target editing in a cell culture model. What...Nat Biotechnol. 2016 Jun 6. PubMed PMID: 27272384 Find plasmids from this publication at Addgene. 2. Kleinstiver...Nature Biotechnology (2016). PubMed PMID: 27347757. Find plasmids from this publication at Addgene. 3. Zetsche....1016/j.cell.2015.09.038. PubMed PMID: 26422227 Find plasmids from this publication at Addgene. 4. Makarova... Guide Pages Check Out MpAgo from the Doudna Lab Find Validated gRNAs ...

...flexible linker is a methyllysine binding domain (chromodomain), which binds selectively to lysine-methylated...like the potential devastation of the palm oil industry through epigenetic effects on the Cover of Nature...maternal copy of the gene is expressed in normal individuals (in a parent-of-origin-specific manner), but ... the histone-derived peptide, the methyllysine binding domain causes a structural change (facilitated ...682-688. PubMed PMID:12404098.  4. Lin, Chi-Wang, Cindy Y. Jao, and Alice Y. Ting. "Genetically encoded ...documentary PBS Additional Resources at Addgene Find the Ting Lab plasmids at Addgene Browse More Fluorescent...

... is the only right way, and Indecisives, who keep changing their minds and have trouble making decisions...categories. In addition, I also learned about two other kinds of difficult supervisors: Hands-off super busy ...work is just one of the hundred things on their minds. When you come to a meeting with a clear agenda,...poor mentors (super-agreeable friendly types, indecisives, and super busy types). If you supervisor ignores...you are advancing career, you will become the independent, assertive, and proactive person that all employers...a Good Mentor Resources on the Addgene Wesbite Find plasmids to Expedite Your Experiments and Achieve...

... pet gives you a clue to help you find the next picture. If you find the next correct photo, you will ...1 below!) You will find five photos this way. At the very last photo, you will find a link to a form where...our recently redesigned blog. 🐕 Your task is to find them, scavenger hunt style. 🔎 Here’s how the scavenger...scavenger hunt works works: You will be trying to find pictures of Addgene pets with Addgene-branded clothing...

...Additional resources on addgene.org Find Optogenetics Plasmids Find Biosensor Plasmids Visit the Fluorescent...multiple other labs have reproduced their EGFP findings since the initial report in 2023.)     ...changes in response to a ~25 mT magnet. Gray regions indicate times when the magnetic field is on. Image reproduced..., in response to a ~10 mT magnet. Gray regions indicate times when the magnetic field is on. Image reproduced...enhanced for future MagLOV-based tools, just as light-induced conformational changes are the basis of many optogenetic...collaborating, the York Lab is eager to hear from you. Find the MagLOV plasmid here!   References and resources...

.... Click on the gene names in the figure below to find plasmids containing that particular gene. ...) and Mark Prescott (Monash University). You can find more plasmids for labeling your subcellular structure...signals not visible to the human eye and are indifferent to subjective color intensity perception. Open...colocalization relies upon the detection of 2 independent fluorescent signals and their potential overlap...muli-color imaging. Furthermore, one must bear in mind that the resolution of light microscopy is limited...interest in a healthy or diseased cell. We hope you find the resources discussed in this article useful for...21209361. PubMed Central PMCID: PMC3074624. 5. Bindels, Daphne S., et al. "mScarlet: a bright monomeric...

...In order to bind DNA, Cas9 and other CRISPR enzymes require a short PAM sequence adjacent to the targeted... researchers have engineered alternative Cas9s binding distinct PAM sequences. Now, Hu et al., working...create point mutations, also has a strict editing window about 15 nt upstream of the PAM. Even if there’...ω-dCas9 could recognize a library PAM, it would induce transcription of gene III, permitting phage propagation...xCas9 activity with wt SpCas9 in mammalian cells, finding xCas9 3.7 displayed about the same editing rate...loci displayed a broad range of PAM sequences, indicating they may be less predictable than SpCas9 off-...Resources on Addgene.org Check out our CRISPR Guide Find CRISPR Plasmids for Mammalian Systems Learn about...

... RNA (gRNA) sequence. Users can design gRNAs to induce the break at a specific genomic site and, using...screen, it is expensive and time consuming to individually generate gRNAs and homology arm repair templates...promote homology directed repair after a CRISPR/Cas9 induced break. Learn more about these constructs and the...for tagging endogenous proteins with an auxin-inducible degron (AID) tag to generate proteins which can...repressors, can be included in the repair template for inducible alteration of protein expression levels. CRISPR...Rapid Protein Depletion in Human Cells by Auxin-Inducible Degron Tagging with Short Homology Donors.” Cell...Resources on Addgene.org Check out the SapTrap Kit Page Find Other Fluorescent Protein Kits Browse the Fluorescent...