We narrowed to 890 results for: IND

...domains tethered to dCas9 often interact with DNA independently of dCas9 targeting (Galonska 2018). This means...importantly, TET proteins affect transcription independent of their enzymatic activity. They interact with...straightforward as expected. TET1 can have many effects independent of dCas9 targeting, including modifying other... This may seem counterintuitive: how can a DNA-binding protein with no enzymatic activity cause demethylation...are key for complete DNA demethylation – tight binding is crucial for dCas9 to interfere with DNMT1 even...02708-5 Hrit J, Goodrich L, Li C, et al. (2018) OGT binds a conserved C-terminal domain of TET1 to regulate...

... However, the molecular and cellular mechanisms behind the immune defects in CDG patients are still poorly...Arg) amino acids more frequently than any other. Indeed, nearly 20% of the single missense mutant alleles...could improve health outcomes for millions of individuals worldwide.Arg is unique among amino acids in ...Rescue occurs through guanidinium cation (Gdm+) binding in the location formerly occupied by the Arg sidechain...) because, at high concentrations (25 mM), Gdm+ binds to and inhibits a potassium channel, causing release...: Baldwin, Enoch, et al. "Generation of ligand binding sites in T4 lysozyme by deficiency-creating substitutions...

...using FRET! Read on to find out more about this amazing imaging technique and find further tips for using...or the activity of a protein in a synapse. What kinds of tools enable us to measure these properties, ...neuronal activity (voltage sensors), to name a few.   Find Many FRET-Based Biosensors on Addgene's Biosensor...PMC18018. 2. Miyawaki, Atsushi, et al. "Fluorescent indicators for Ca2+ based on green fluorescent proteins ...Biosensors Additional Resources on Addgene.org Find FRET Pairs Check Out Fluorescent Protein Biosensors...

...our Hot Plasmids articles.  Here's what you'll find in this post: Peptide-assisted genome editing for...Adapted from Zhang et al. 2023 with permission.   Find Cas-PAGE plasmids here!   Zhang, Z., et al. (2023...on mStayGold inspired and informed this section.  Find mStayGold plasmids at Addgene here!   Ando, R.,... Goeden, et al. 2023, under CC-BY license.   Find the AAV.CAP-Mac plasmid here!   Chuapoco, M.R.,...from Chen et al (2023) under CC-BY license.   Find AAV1-X1 plasmid here! Chen, X., et al. (2023). Functional...has been widely used in a variety of applications. Find anti-DYKDDDDK recombinant antibodies here!...

...yourself: is it clear how novel my findings will be and why these findings will be worth knowing? This can...tricky, as most of our research builds on ideas and findings from our own or other groups, but your work must...to know what we are doing and why. I always ask kindly friends and family to read my non-expert paragraph...back at my successful applications, I can't always find something that made them better than my unsuccessful...up an ELISA kit company on whose website you can find some great information on ELISA troubleshooting ...

...SSRs bind DNA at target sites to induce site specific recombination events: Cre recombinase binds loxP...the Cre-loxP recombination system can be used to induce site-specific recombination events, and that the...loxP sites, while FLP binds FRT sites. When a DNA sequence is flanked by target sites (floxed) in opposing...PMID: 2286602. PubMed Central PMCID: PMC3406316. Find FLEx plasmids from these Addgene depositors: Ed...

...site selection by TnsD, a sequence-specific DNA binding protein. Selection removes donor bacteria and recipient...subtilis to the bacterium of interest following IPTG induction of conjugation genes and bi-parental mating. Following...and colleagues used Mobile-CRISPRi to knockdown individual genes in several bacteria associated with human...measured after 6 and 12 generations with or without induction of CRISPRi knockdown. As expected, bacteria expressing...move Mobile-CRISPRi into your bacteria of interest? Find Mobile-CRISPRi plasmids at Addgene! References...Addgene.org Browse our CRISPR interference plasmids Find bacterial CRISPR plasmids Read our CRISPR guide ...

...this kind of animal models. Start using adenoviral vectors with your CRISPR/Cas9 research! To find more...cells, Cas9 machinery is transiently expressed and binds to target DNA. In the final step of targeted gene...region. The CRISPR/Cas9-derived RNA-guided nuclease-induced gene disruption frequencies in the various cell...looking for a broader range of CRISPRs plasmid tools, find more plasmids, CRISPR technology guides, FAQs, and...delivery of RNA-guided CRISPR/Cas9 nuclease complexes induces targeted mutagenesis in a diverse array of human...

...traditional restriction cloning. Sequence and Ligation Independent Cloning (SLIC) could also be a great choice, ...non-recombinant vectors from propagating. The ligation-independent methods like Gibson and SLIC are also quite efficient...you some tips for your next project. But keep in mind that there are many other cloning methods - including...don’t be afraid to explore multiple approaches to find out which one works best for you. Happy cloning ...simplified, purification-free, sequence-and ligation-independent PCR cloning method." BMC biotechnology 11.1 (...Addgene.org0 Check out protocols for plasmid cloning Find vector backbones Read our molecular biology reference...

... Is it an evolutionarily conserved gene? Can you find an ortholog in the Drosophila genome? Continue reading...describe how you can make your own mutant flies. Find Drosophila Resources at Addgene The birds and the...to help keep your research moving. Below, you’ll find some great introductory resources. As you dive deeper... deeper into Drosophila research, you’ll find more resources that may be more tailored to your field of.... The Bloomington Drosophila Stock Center at Indiana University is another tremendous resource for the...Drosophila Learn about Cre-Lox Resources on Addgene.org Find Drosophila Plamids Drosophila CRISPR Resources Deposit...

...whether as part of protein fusions or expressed independently) originally fluoresce green or blue and, when... (green or blue) within a population of timer indicates the age of the pool: the higher the ratio, the...is dependent on the total expression time but independent of protein concentration. Calibration curves ... red/green fluorescence falls within the linear window for the theoretical timer shown in Figure 1B, you...cell structure, etc). FP timers should therefore find use in studies of animal development where events...Your Fluorescent Protein Resources at Addgene.org Find Fluorescent Protein Timers Browse All Fluorescent...

... and an extension sequence containing a primer binding site and a reverse transcription template (RTT)...high-throughput libraries My labmates and I wanted to find a way to predict the efficiencies of pegRNA designs...the RTT overhang, the GC content of the primer binding site, and many more. Once we had our datasets with...experiments Prediction models only become useful once they find their way into the hands of users. For this reason...resources References Akhtar, W., De Jong, J., Pindyurin, A. V., Pagie, L., Meuleman, W., De Ridder, J....B. (2021). Impact of chromatin context on Cas9-induced DNA double-strand break repair pathway balance....

...following techniques to increase plasmid yield. Keep in mind that you might need to use multiple techniques for...that a higher volume of lysate passed over the binding matrix could clog the matrix when performing filter...determined for each plasmid/strain combination individually, based on determining the cell density or by...Measuring the optical density of the culture can indicate the number of cells present in the given volume... standard 37℃. This plasmid feature should be indicated in the plasmid information or growth strain instructions...coli Strains Additional Resources on Addgene.org Find Info on How to Quantify Your DNA Learn to Do a Bacterial...

... switch to Golden Gate Cloning easier because individual labs will not need to ‘domesticate’ all the basic... they were developed mostly with model plants in mind, Arabidopsis and Nicotiana benthamiana. N. benthamiana...Golden Gate cloning system especially with plants in mind. When it’s ready, I’ll make those available on my... up making 30 constructs. They’ve never done any kind of cloning and they don’t have any prejudice about... active on Twitter. Sometimes it’s a good way to find out what’s happening in the office next to you. ...remarkable number of female scientists whose tweets I find really inspirational. They share experiences, articles...

...proficiency.  What kind of content are you sharing?  Sometimes your subject matter will drive the kinds of communications... I want to put to rest a common misconception. I find personal bias often unintentionally plays a big ...My dad, on the other hand, mastered the art of finding how-to videos on YouTube long before he learned...audience already is.  You’ll also want to keep in mind that access and availability to resources (including...embark on a project, do your best to understand what kind of resources - including your time - will be required...

...protein (FP) is split into two fragments that individually lack fluorescence. When complementary FP fragments...resources in less genetically tractable organisms lags behind. BiFC can also be used to screen for small-molecule...carry out because it relies on a single BiFC pair. Find BiFC plasmids for your experiment at Addgene! How... the random association between FP fragments independent of PPIs. Overexpression can be assessed by comparing...fluorescence due to the random association between FPs independent of PPIs. Simply compare the fluorescence intensity... introduction to FRET Resources on Addgene.org Find fluorescent protein plasmids and resources Visit...

...of genome engineering via CRISPR.  Though the individual topics are not surprising, the real value here...Science,” he describes the now infamous tale of Rosalind Franklin and her essential-but-unlauded role in...quieter, more reserved Franklins and Prashers will find it increasingly difficult to compete (for funding... quite a walk down memory lane for me. A brief window in time “The most obvious future consequence of... but that’s unlikely to be foremost in readers’ minds. Six months ago I would have nodded my head knowingly...book left them wanting to learn more. There are individual paragraphs that may have warranted their own ...

...expressing a protein that directly binds to zeocin, preventing it from binding DNA. Zeocin is an all-in-one ...a way to produce more of the plasmid, isolate individual plasmids, or express a gene contained in a plasmid...member of the bleomycin family of antibiotics, which induce cell death by intercalating into DNA and generating...genotoxic when not 100% inhibited, meaning it will induce mutations in the host/plasmid DNA during selection...the Sh ble gene does not fully suppress Zeocin-induced DNA cleavage in human cells. Mutagenesis, 20(2)...

... determines how long and precise we can track individual molecules. Poor photon budgets are a widespread... synthetic fluorophore-contianing “ligand” that binds to the tag. Like most good ideas in biological imaging...proteins (FAPs) – These modified antibody fragments bind and enhance small molecule fluorogens (Figure 1e... encoded tag is required as the small-molecule binding motif targets endogenous protein (Figure 1f). ...when free in solution, but high fluorescence upon binding to its cognate protein. In some cases this property...Request Plasmids from the Michael Davidson Collection Find Fluorescent Protein Kits ...

...Plant Plasmids and Resources page Find synthetic biology plant plasmids Find CRISPR plant plasmids ...plant. A common method for silencing genes Virus induced gene silencing (VIGS) is a transient technique ...subcellular localization of your protein of interest. Find all plasmids from the Lee-Parsons lab. Test different...PMC6585625. Unver, Turgay, and Hikmet Budak. "Virus-induced gene silencing, a post transcriptional gene silencing...Addgene blog Read all of our plant biology blog posts Find tips for Arabidopsis transformation Read our Plasmids...

... above showed successful target modification, indicating that AAV is a good delivery vehicle for Cas9....cleavage activity in mammalian cells. SaCas9 produced indels at a similar efficiency to SpCas9, leading the ...tested two genes in mice. For both genes, they saw indel formation and phenotypic changes as early as 1 week...packaged in AAV.   Split AAV approaches  While finding smaller Cas9 orthologs works for some applications... in two pieces and package each piece into an individual AAV. When a cell is transduced by both AAVs, ...Resources on Addgene.org Check out our CRISPR Guide Find CRISPR Plasmids ...

... To create the eGFP reporters the Harris lab individually mutated three codons in eGFP to eliminate fluorescence...capability of the seven human APOBEC3 proteins, finding that APOBEC3A and APOBEC3bctd are the most efficient...using a base-editor (C>T conversion) they could induce the translation of any detectable protein, not ...contained the cytidine base editor exhibited robust induction of GFP fluorescence confirming the efficacy of... universal, the Dow lab successfully used GO to induce the translation of an array of different reporters...Resources on Addgene.org Browse the CRISPR guide Find CRISPR plasmids for your research ...

...et al. 2010), inducible promoters with different degrees of stringency and inducibility (Toh et al. 2023... replication initiation factors and proteins independent of the host. Host range depends on whether the... vary in DnaA-box and iteron structure, host-independent   Building broad host range recombinant...2023), and additional cloning sites (Keen 1988).  Find broad host range recombinant vectors at Addgene!... (2023). Development of constitutive and IPTG-inducible integron promoter-based expression systems for...

...Use the links below to find Gateway plasmids for your organisms of interest: Find Gateway cloning plasmids...carry different antibiotic resistance markers (indicated here by plasmid color), allowing you to easily...pLenti CMV Puro DEST (w118-1) or the doxycycline-inducible pLIX_403. The chosen attR destination vector will...fragments, cDNA or Genomic DNA and is available for all kind of organisms from mammals to E. coli. Glossary of...Quickly Verify Your Clones Resources on Addgene.org Find Plasmid Cloning Protocols Watch Our Diagnostic Restriction...

...forum discussions about a wide variety of topics. Individual forums are hosted on subreddits (r/”topic”) which...yourself and your work out to other scientists who will find it useful, I highly recommend doing an AMA! Before...something we are, not something we do. We have to find a way to keep doing science to be happy. The happy... I said in the AMA, it’s incredibly important to find good mentors and incentivize good mentorship: AMA...get to speak publically about your work (you can find my tips for a perfect science talk here). Finally...the features I need to save, organize and easily find the resources that I come across on the internet...

...different or novel tools and assumptions to reveal findings that the originator was oblivious to, no matter...of the experiment would require access to each individual video. To PLOS’s credit, the organization subsequently...recognize that storing data in a sharable form is indeed a significant effort. Our standards of data recording...recording are highly variable and usually left to individual researchers. Yet accurate and comprehensible ...appears to leave the scope of data to be shared to individual authors. Minimally, raw but sufficiently annotated...charges of elitism and for scientists to be hiding behind our jargon and esoterica. Public trust in and funding...

...from the termite hindgut. Want to level up your or your class’s streak plate skills? Find the chromoprotein...Resources on Addgene.org Find fluorescent protein plasmids at Addgene Find chromoprotein plasmids at ...the students to separate out colonies from each individual strain using the streak plate technique. Time...

...these types of conflicts is important for the individuals involved, and also for the labs, organizations...they were taking care of that”, because that can indicate confusion around... Ownership: Misunderstandings...tools that can come in handy: Empathy: If you often find yourself in confrontational situations with others...the age of five. But it does mean that even if you find someone annoying or disagreeable, you recognize ...Definitely not. But it can help, and if and when you find yourself transferring this skill to another job,...for avoiding, competing, or accommodating, if you find yourself gravitating most frequently towards these...

...copies of the transgene that will recombine independently, cells may acquire one of a variety of colors...extremely versatile technology as recombination can be induced in developing fish via Cre protein microinjection...microinjection or crossing of Zebrabow animals to an inducible Cre line. The Schier group developed Zebrabow as ...powerful tool for understanding cell proliferation. Inducible XFP expression in the Ubow mouse Another murine...CFP, and YFP. After crossing onto a tamoxifen-inducible ubiquitin background, each cell in the mouse that...Köhler, Corinna, et al. "Mouse cutaneous melanoma induced by mutant BRaf arises from expansion and dedifferentiation...

..., 2008). Shortly after though, three groups independently tried reproducing these results and determined... and part B. Each piece of the gene is then independently packaged into an AAV. When a cell is co-infected...Viral Vector Packaging Capacity in a Transgene-independent Manner. Molecular Therapy 16:124–130 . https:...AAV Transductifon Is Mediated via a DNA-PKcs-independent, Rad51C-dependent Repair Pathway. Molecular Therapy...ddv386 Additional resources on the Addgene blog: Find tips for a first time AAV user Learn about how AAV...on Addgene.org: Check out the Addgene AAV guide Find ready-to-use AAV preps from Addgene's viral service...

...single-component systems take advantage of BcLOV4’s light-induced binding of the plasma membrane. In previous publications...light BcLOV4 undergoes a conformational change and binds the plasma membrane under blue light CRY2-CIB1 heterodimerizes...limitations (Figure 1). Figure 1: Opto-katanin induces fast, localized microtubule severing. A) Schematic...substantial PA accumulates in the plasma membranes, indicated by the PA probe colocalizing with SuperPLDx30....CRY2-CIB, and other optogenetic modules? Or want to find more optogenetic tools in our collection? Check ...

...complexity inside the cell or on its surface. Bear in mind, the voltage setting of the laser impacts the magnitude...While flow cytometry can be used for virtually all kinds of experiments and research fields, for any experiment...After that, the next gates fully depend on your individual experiment. Practical example: lymphocytes in...cellular debris and other larger cells. Note, we don’t mind the warning message at the top, as our cells of ...strategy, I can, of course, also draw multiple individual gates around the populations of interest (Figure...of a quadrant gate, I am free to use multiple individual gates. Note that in this case, square gates were...

...homology-directed repair (HDR) of a CRISPR/Cas9 induced double-stranded break. This site-directed genome...rendering the resulting construct resistant to Cas9 induced cleavage. Summary of Site Directed Mutagenesis ...is removed by DpnI restriction digestion leaving behind only the mutagenized plasmid. Although it is commonly...reaction. Ideally, your primers should be free of palindromic and repetitive sequences, but if present, a minor...similar approach can be used to identify Cas9/CRISPR-induced genome modifications. Occasionally, multimerization...Addgene.org Browse Our Molecular Biology Protocols Find Empty Vectors for Your Next Cloning Experiment ...

...on Addgene.org Find Synthetic Biology Plasmids Find Bacterial Expression Systems Find Plasmids for Protein...optimized for parameters such as induction temperature and length of induction, which will vary between different...gels containing Phos-Tag, a small molecule that binds specifically to phosphate moieties and promotes ...

...Advances in Cre-lox recombination have included inducible Cre, such as the tamoxifen-responsive CreERT, ...single-cell labeling is necessary to distinguish individual cells. The Brainbow-1.0 construct contains three...mice.) The combinatorial effect of multiple, independent recombination events leads to a rainbow of colors... with the vast array of colors needed to mark individual neurons, the system also suffered from a number...they screened a variety of fluorescent proteins to find those with ideal characteristics (low aggregation...Check out Brainbow plasmids available from Addgene! Find Brainbow Plasmids @Addgene: Brainbow 1.0, 1.1, ...

...science funding, limitations to facilities and indirect costs, and increased tariffs present an unprecedented... of the cost of creating and verifying them independently — particularly valuable when budgets are constrained...preparations, and antibody production that labs may find increasingly difficult to maintain or access. These...applications, addressing the new funding structures and indirect cost caps more efficiently. Offering free educational... seeking and expanding our collaborations with industry and public and private partners to create additional...

...similarly to their untagged counterparts. These findings motivated Georgieva et al. to develop new tags...from non-vertebrate systems. Preliminary testing indicated that inntags IT5, IT6, and IT10 expressed well...on these parameters. IT5 and IT6 displayed good binding efficiency in both immunofluorescence and immunoprecipitation...for both applications. To prove that inntags are indeed innocuous, Georgieva et al. created tagged versions...detrimental effects on cell size and growth rate, indicating that they compromised protein-protein interactions...

...the toolbox, with the first paper identifying a binding receptor published this year (Dhungel et al., 2025...for your experiment? Use Addgene’s AAV Data Hub to find experimental reports on commonly used vector cargo...Serotype 4 (AAV4) and AAV5 Both Require Sialic Acid Binding for Hemagglutination and Efficient Transduction...Viswanathan, S., Gaj, T., Lavzin, M., Ritola, K. D., Lindo, S., Michael, S., Kuleshova, E., Ojala, D., Huang..., M. J., Chiorini, J. A., & Zabner, J. (2001). Binding of adeno-associated virus type 5 to 2,3-linked ...α2,6 N-Linked Sialic Acids Facilitate Efficient Binding and Transduction by Adeno-Associated Virus Types...

...protection against infectious agents. Several enzymes behind this immunity have already been discovered and ...coli bacteria strain. They found that Cas14 could bind and cleave a targeted sequence of ssDNA. Unlike ...cleavage, the authors showed that Cas14 could indiscriminately cleave ssDNA in trans like Cas13 and Cas12...of dsDNA and RNA in trans. Fluorescent signal indicates the presence of this viral sequence in the assay...our CRISPR cheat sheet Resources on Addgene.org Find the Cas14 plasmids here Checkout our CRISPR guide...

...Advisor/PI who will also be a good mentor? Where can I find a Mentor? What is this I hear about “group” or “...“A good mentor has your goals/needs/situation in mind, rather than (or as well as) their own.” “Someone...mentors/advisors like Yoda? If so, you might want to find additional sources of wisdom. While somewhat lighthearted...on the blog “Tenure, She Wrote” is titled, “What Kind of Mentor Do I Want To Be?” In this excellent essay...the northeast, writes a thoughtful treatise on the kind of advisor/mentor she wants to be for her new lab...