We narrowed to 801 results for: DES

... While it is certainly possible to select for desirable traits using traditional plant breeding approaches...modification of known or suspected genes that regulate a desired phenotype. In fact, CRISPR has already been used...wheat, mushroom, and rice (Khatodia et al., 2016). Despite the almost universal functionality of the CRISPR...genes in plants using the same general experimental design principles developed in other model organisms (...nanostructures (Zhang et al., 2019) or cell-penetrating peptides (Santana et al., 2020) and plants viruses (barley... through Addgene. In addition to the plasmids described above, Addgene carries several useful CRISPR toolkits...CRISPR Multiplexing Tools Validate Your Genome Edit Design Your gRNA Additional Resources on Addgene.org ...

...comments section.  FX cloning kit This kit is designed to be used with Type IIS restriction enzymes and...this kit have been used to purify a variety of peptides including antibiotic efflux pumps and ankyrin ...et al. "A transcription blocker isolated from a designed repeat protein combinatorial library by in vivo...followed by leader sequences (encoding small leader peptides) downstream of which you can clone your gene of...Shine-Delgarno sequences used in this so-called bicistronic design (BCD). The 49 vectors in this kit can be used to...(2015): 31-37. pCri system The Gomis-Ruth lab designed the pCri system to simplify the protein purification...for cloning. Furthermore, some plasmids contain peptides to facilitate protein folding and/or protease ...

...controls and be able to use this information to design thoughtfully and thoroughly controlled experiments...is generally a sample or group that will have a desired response. For example, if you are studying the ...sample, the antibody binds specifically to its desired target (purple) and nonspecifically through the...The Preimmune serum control will not bind to the desired target but will bind nonspecifically through the...the Fc, allowing you to detect this undesirable interaction. When non-specific interactions are suspected...nonspecifically with positively charged groups, such as aldehydes in fixed samples. To address this, block positively... will be a useful starting point for you when designing rigorous experiments. While this blog touched ...

...most clinically-relevant proteins are also the hardest to study. One common strategy is protein overexpression...compatible with both strong and weak promoters, and even modest endogenous pools generate high signal-to-background...introduce mutations. This vector-based use of HiBiT provides a means to quickly answer research questions before...the code of 'undruggable' targets Researchers can design drugs to bind to well-defined ligand-binding pockets...mechanistic drug responses (Kamio et al., 2025). They designed HiBiT-tagged TP53 constructs, one with an R175H... links p53 mutations to treatment outcomes and provides a HiBiT-enabled framework for quantifying variant...many difficult-to-study protein targets, HiBiT provides an excellent solution compatible with high throughput...

...Glutamate-Sensing Fluorescent Reporter (iGluSnFR) designed and improved by the Loren Looger and Kaspar Podgorski...Aggarwal et al., 2025).In this latest work, the team designed and screened a large library of iGluSnFR3 mutants... HA-, and EGFP-expressing lentiviral backbones designed right here at Addgene. Here's what makes these... stable cell line creation a breeze. Modular by design: Key elements (promoter, tag, polyA site) are flanked... plasmids for flexibility and speed, so you can design, test, and innovate faster than ever. Plus, we'...have a two-for-one solution: TurboCas!TurboCas provides the genomic targeting power of dCas9 with the ....2024.11.007. Branon, T. C., Bosch, J. A., Sanchez, A. D., Udeshi, N. D., Svinkina, T., Carr, S. A., Feldman, J. ...

... Roth BL (2015) DREADDs (Designer Receptors Exclusively Activated by Designer Drugs): Chemogenetic Tools...less invasive and more flexible as it relies on designer ligands that can be simply introduced by injection...lab and Takafumi Minamimoto’s lab known as deschloroclozapine (DCZ) has been developed to meet these requirements...should be carefully controlled for in experimental design.   Optogenetics Chemogenetics Timing Fast ...Find our chemogenetics and optogenetics science guides ...

...CRISPR inhibition (CRISPRi). A) Lentiviral vector design. B) Strategy for leak-free transcriptional control...Organoid EasyTag workflow combines thoughtfully designed plasmid components with Fluorescence Activated... heavy optimization. Claudia Vickers’s lab has designed novel SL fluorescent biosensors by integrating... rather than on the fluorescence intensity, it provides a direct and quantitative detection of SLs. The...biosensors could have a major economic impact.   Design and mechanism of action of the receptor-based SL...targets all mouse genes with 10 gRNAs per gene. The guides are optimized to maximize on-target cleavage and...

...tagging method that labels cells with heritable barcodes. Barcodes consist of an 8bp variable region and a 6...Direct reprogramming describes the process where differentiated cells are turned into a cell type of ...with the majority of cells failing to develop the desired identity. The development of single cell technologies...Morris Lab at Washington University in St. Louis provides a combinatorial indexing strategy that simultaneously...towards successful iEP reprogramming. MettI7a1 encodes an uncharacterized putative methyltransferase. ...

... been described in this blog. If your data does not meet the assumptions for the tests described above...experimental groups, but how do you determine if they deserve that oh-so coveted asterisk of significance? You...referred to as two-sample or independent, but these designations all refer to the same test. On some occasions...Correction and the Benjamini-Hochberg procedure are designed to reduce false positive rates and control for...error rate than the multiple comparison tests described above. Violated assumptions and statistics Statistical... making your statistical analysis plan when you design your experiment, instead of after you’ve already...

...to a wide range of industries and is a highly desirable quality sought after by employers. These skills...correctly the first time. These skills are very desirable to employers because they save time and money ...Meteorologist Journal Editor Textile Technologist/Designer     Fieldwork and traveling Many people are...employers. Increasingly, computer skills are highly desired, as roles are becoming more reliant on technology...(Python, R, SAS, HTML, Matlab etc) are highly desirable in a range of roles, including: Science Reporter...find answers to include: Will you be sitting at a desk all day or be on your feet constantly in a lab? ...

...or FP) is one of the most important steps in the design of an imaging experiment.  The choice is non-trivial...fusion protein, etc...).  There are many online guides – including an excellent series of posts by Joachim...information about fluorescent proteins.  It was designed with the goal of encouraging user contributions...while also maintaining a relatively strict database design (schema).  Registered users can submit new fluorescent...risk of inaccurate data submissions, so FPbase includes a versioning system that allows users to see when...for, the Table of Fluorescent Protein Properties provides a master list of all proteins in the database,...

... lacks context unless it links to a verified description of that reagent online. Furthermore, if the link... link between the catalog number and reagent description is lost (e.g. a supplier is bought out by another...directly to the original article online. It is also designed to be persistent and exist online indefinitely...specific information about the plasmid: name, description, how the plasmid was generated, validated sequence...authors comply with journal policies, Addgene now provides an RRID and a resolvable compact identifier for...you directly to the lenti-Cas9-VQR plasmid page. Despite their differences, both of these PID formats for...

... has precluded widespread adoption of the technology. In 2012, Saito et al. described the first Nano-...molecular and cell biologists over the last two decades, but their use can be limited by a few caveats....hours! The latest enhanced Nano-lanterns (eNL) described in Suzuki et al. improve the brightness of previous...substrate furimazine. In addition to the previously described applications, the enhanced signal of eNL allows...autofluorescence and a high signal-to-noise ratio. Despite the many benefits of Nano-lantern technology, there...

...infectivity): The use of the VSV-G envelope provides the widest tropism or range of cells a virus can infect...The widespread use of γ-retroviruses (gamma-retroviruses) in cancer and stem cell research has prompted...lentiviral packaging methods. The three components described above (envelope, packaging, and transfer) are ...into the 293T packaging cell line. This system provides the greatest flexibility to pseudotype γ-retrovirus...nomenclature of the plasmid backbone is meant to be descriptive of the components found within the backbone. ...

...is not enough on its own to help you find your desired plasmid. In many cases, you will need a screening...you will need to sort through them to find the desired clone. Do your selection, then screen Why add a...what about identifying clones that contain your desired insert? This is where a  screen comes in. Let’s...blue-white screen, which relies on the lacZ gene. lacZ encodes the enzyme 𝛽-galactosidase which can hydrolyse...Sanger sequencing determines the precise order of nucleotides within a DNA molecule, in this case a plasmid... of the information you need, so you'll want to design a custom primer. Because Sanger sequencing can ...

...created SRIRACCHA in 2017 to enrich and identify desired CRISPR mutants (Wen et al., 2018). SRIRACCHA uses...constructs for the reporter to work and to make the desired genome edit:  A donor plasmid that contains a ...OH-T and doxycycline. A gRNA that targets your desired gene (whose sequence is also found in the reporter...well. The ratio of RFP to GFP expressing cells provides a real-time read out of Cas9 activity.  By using...integrated GFP reporter leaving you with just your desired genomic edit.  Find SRIRACCHA plasmids at Addgene...DSB is repaired by NHEJ which depending on the nucleotides inserted or deleted, can result in an in-frame...

...educational resources — including blog posts, protocols, guides, and collections — is a lot of material to sift... launched earlier in the year, as well as our redesigned homepage and our new search tool, are the results...a few of the strategies we used in our software design. In each case, we leveraged the team’s knowledge...per resource category to expose as filters and designed the search engine to return only the filters that... bits of information for a given resource, and designed the search engine to weight search results differently...'s new search Read to learn more about why we redesigned our homepage and mascot ...

...assessed for the desired parameter. A droplet containing a cell that is positive for a desired parameter is...their observations. However, while flow cytometry provides analysis at the cellular level, it cannot be used...degree of scatter is captured by detectors and provides useful information about the cell. Light that ... traveling is called forward scatter (FSC) and provides information about the relative size of the cell...initially traveling is called side scatter (SSC) and provides information about the cell’s complexity. A cell...side scatter (SSC). Users can gate around their desired cell population for further analysis. In this example...

... molecule degraders” recruit the proteasome to destroy targeted proteins. We previously developed small...locus-specific knock-in of FKBP12F36V. Detailed protocols describing the use of these plasmids are available here....Nabet et al., Nature Chemical Biology additionally provides detailed information on cloning procedures used...degradation, and lead author on the recent paper describing the dTAG technology. Follow him on twitter @behnamnabet... 29581585. This reference contains the full description of the dTAG system. The dTAG system was employed...Conditional Mouse Models CRISPR 101: Cas9 Nickase Design and Homology Directed Repair Plasmids for endogenous...

...into agriculture as species-specific pesticides and insecticides, an advancement over previous chemical...prone to degradation – those are the words that describe RNA! Double stranded? Well, that’s just for DNA...also known as RNAi, are short, dsRNA molecules designed to degrade mRNA transcripts. RNAi activates the...sequence is found (typically RNA that the RNAi is designed against), RISC can repress translation and/or ... and repetitive sequence. All of the R-loops described above play distinct roles in promoting cellular...

... could participate. Despite the group not having a public website, all the slides, handouts, and resources...science or even to discuss potential solutions. Despite all the talk, not much was being done to address...practices could be an actionable way to tackle widespread irreproducibility. Luckily, there were other ...thinking about reproducibility and shared similar desires to take action. Thanks to Teytelman’s vast network...practices could be an actionable way to address the widespread issue of irreproducibility. Use your network...

...approaches to gender equality over the past several decades: “Fix the Numbers of Women” focuses on increasing...-art methods of sex and gender analysis and 2) provides case studies with concrete examples of how gender...using both sexes in basic research is excellent design), but they used all female stem cells—this was ...environmental factors. How to include sex in experimental design Recognizing the importance of analyzing sex, the... above, another excellent resource for research design is Kalpit Shah et al., “Do you know the sex of ...Corporations need to integrate these insights into product design. But eyes have been opened—and we cannot return...

...monitored over time by sequencing the barcodes in the population (all barcodes can be amplified using the same...with roughly 73 million semi-random, 30 bp DNA barcodes that are integrated into target cells' genomes...ClonTracer system, lentiviruses containing these barcodes infect the target cell population at low multiplicity...replicate populations, they found that similar barcodes consistently made up the majority of the resistant...resistant populations. In fact, 88% of the barcodes within the resistant populations were shared by at least...

...repository that will help epigenetic researchers custom design their own engineering plasmid tools in about a ...cloning, the Haynes lab’s Protocols.io protocol describes an elegant system that allows for the assembly... cells that make up the human body and G-baToN provides versibility in tracking and modulating the interactions... inhibitor response 1 (TIR1) expressed under a desired promoter with a specific spatiotemporal expression...interests via CRISPR techniques. This tool-kit includes an array of FPs as well as additional useful tags...libraries encode target sites flanked by randomized nucleotides in place of a protospacer adjacent motif (PAM...

...of 3’UTRs, the Arizona State University (ASU) Biodesign institute developed a 3’UTR database specifically...specifically for C. elegans. This database provides detailed information about C. elegans 3’UTRs structures, ...imaging processing. The combination of these methods provides unique opportunities to develop computational ...Addgene In addition to the wealth of open resources described above, there were many plasmid tools presented...self-excise, and covalently ligate their flanking peptides together (Wang et al., 2018).” With the split ...continuously curated as we continue to receive plasmids designed for the worm so check back frequently for what...

...high quality DNA. Figure 1 illustrates the three grades of DNA you may need based on their relative quality...components (i.e. buffers, columns, or beads) are designed to work together — no mixing and matching!   ...bacterial culture. Plasmid purification kits are designed to accommodate specific volume ranges of culture...column-based miniprep kit for example. These kits are designed to handle ~1–5 mL of liquid culture. Using a higher.... To help combat this, some manufacturers have designed eco-friendly DNA purification kits by reducing...Milllipore Sigma’s GenElute™ kits were all specifically designed with environmental sustainability in mind (Table... kits from a variety of manufacturers. It also includes a rough comparison of list price between these...

...genetic locus when complexed with an appropriately designed small RNA, or guide RNA (gRNA). The gRNA conventionally...exploited to either inactivate a gene or introduce a desired genetic alteration. Listen to Our Podcast Interview... truncated such that it exhibits 15 or fewer nucleotides of complementarity to a target site, Cas9’s DNA...truncated gRNAs Importantly, the use of truncated guides has largely been demonstrated to result in decreased.... An additional benefit of the use of truncated guides is that they can be delivered into systems that...Cas9 species. Within a single cell, full-length guides targeting one set of genetic loci can be introduced...

...amazing project. You’ve written your protocols, designed your experiments, and prepared your reagents. ...following cloning methodologies have been extensively described in other articles; so if you are interested in...homology. You might also consider FastCloning, first described by Chaokun Li and colleagues. Here you simply ...-generated insert. When mixed, topoisomerase I provides both the endonuclease and ligase function while...restriction enzymes. In Golden Gate cloning, the desired end construct does not contain the type IIS recognition...This feature biases the reaction in favor of the desired assembly to make the process close to 100% efficient...

...insertion site, the corresponding gRNA might be undesirable due to potential off-target sites or low predicted..., resulting in lower overall genome frequency. Despite the progress we’ve made, there is still a need ...bacterial polymerase subunit ω. This construct is designed to activate transcription from the accessory plasmid... stringency and promoting survival of only the broadest-targeting variants. For the final round of evolution...spectinomycin resistance. xCas9 3.7 showed the broadest PAM range, recognizing NG, NNG, GAT, and CAA PAMs...scaffold. With an NGG PAM, xCas9 3.7-VPR constructs modestly outperformed SpCas9-VPR in activating transcription...

...degradation of the tagged protein. Degrons are destabilizing domains which will be identified by various ...proteasome for degradation. Like several other tags described here, the dTAG uses a small molecule, a tag, and...protein for imaging. Given its popularity and widespread usage, a proteolysis-targeting chimera (PROTAC... The auxin-inducible degron (AID) system is the oldest system reviewed here. The AID system relies on ...Thankfully, this is a hot topic with many resources! Bondeson et al. (2022) reviews all the tags discussed here...choice!   References and Resources References Bondeson, D.P., Mullin-Bernstein, Z., Oliver, S. et al....

...Snapp. Stop using EGFP/GFP for fusion proteins! Despite multiple studies in high profile journal articles...world examples of these situations in cells are described below (See Figure 1).   Figure 1: Each...of environmental changes that the biosensor is designed to detect resulting in false positives [2, 4]....Addgene has many of these constructs and they are designated mEGFP, mVenus, etc. A word of caution is needed... not necessarily monomeric, even if they were described as monomeric in a publication. The gold standard... [2]. Some affinity assays used in the papers describing the FPs above including molecular sizing columns...

... to imagine designing 10-gene systems with quantitative precision. There are few designers of biological...made news in March with a publication in Science describing the development of “transcriptors,” transistor-like... and observed that we humans find it useful to describe and interact with the world using words like OR...The future of synthetic biology Addgene: You've described the pieces in these kits as the beginnings of ...laying down a challenge to its users and to the design community: If you can begin to reliably put together...

... crRNAs for Cpf1-mediated cleavage are 42-44 nucleotides in length, about the same size as Cas9’s, but...SpCas9. Since Cpf1 doesn’t require a tracrRNA, crRNA guides are only ~42 nt long. Direct synthesis of these... than that of the ~100 nt crRNA/tracrRNA hybrid guides needed for Cas9 function. Since both Cpf1 and its... endjoining (NHEJ). Cas9-mediated NHEJ usually destroys the PAM site due to its proximity to the cleavage...chance” mechanism might improve the frequency of desired HDR edits, but the possibility has not yet been...to examine Cpf1 off-target editing. First, they designed gRNAs with single and double mismatches throughout...

...Cas13a as a diagnostic tool. A pool of DNA or RNA nucleotides containing a sequence of interest (red) is amplified...Reverse-transcription RPA (RT-RPA), respectively. Amplified nucleotides are combined with Cas 13a in complex with crRNA...the target sequence is present in the pool of nucleotides, the nonspecific RNAse activity of Cas13a becomes... sequence is present in the original pool of nucleotides.   What are the potential applications of...can mediate cleavage with spacers from 20-28 nucleotides in length and does not require a specific PFS... the collateral RNA cleavage response is well described in bacterial cells, LwaCas13a does not display...

...iPSCs. The generation of human iPSCs (hiPSCs) now provides an opportunity to develop and use patient-specific...for reprogramming purposes. In this post, I’ll describe an episomal vector system that increases reprogramming...the CytoTune™-iPS 2.1 Sendai Reprogramming Kit designed to generate clinical grade hiPSCs is sold at $11,000...replication, OriP and the EBNA-1 sequence that encodes for the DNA binding protein EBNA-1 [7]. These two...reported efforts. The pCXLE episomal system thus provides a practical way to generate transgene-free, virus-free...factors they encode Episomal Plasmid Addgene ID# Encodes pCXLE-hSK 27078 SOX2 and KLF4 pCXLE-hUL 27080...

...site, and titer. Addgene’s Viral Vector Service Besides being complex to use, viral vectors are also difficult...which include calcium and voltage sensors, are the broadest category of viral vector tools, which may help... for vectors in the “controls” category, which includes things like viral vectors that deliver GFP. ...most requested serotypes (Figure 4), our catalog includes vectors in a variety of serotypes, including the...early engagement with UPenn to our launch of the redesigned Data Hub in 2022. It’s influenced the way we ...viral vector educational resources, from eBooks to guides to protocols, which was why I was so excited to...

...their BioRxiv post, the authors generated and described several improved genetically encoded fluorescent...Takayuki Iwaki, Kimiko Hara, and Kazuo Umemura have described a simple method which alleviates the inclusion...contains the signal peptide of MBP, followed by the desired nanobody (NB) sequence, an AviTAG, and 6xHis tag...expression can vary significantly depending on transgene design and delivery. To overcome these variations in human...Addgene The MoClo CRISPR/Cas Toolkit for plants includes 95 plasmids consisting of CRISPR/Cas nucleases... uses CRISPRi to control gene expression. This includes a CRISPRi-based synthetic oscillator, bistable...

...released a major redesign of how search works on our site. For Addgenies, development includes everything from...experience and the overall soundness of the software design. Each of us brings slightly different cognitive...Addgene. Another advantage of Scrum is that it provides transparency to everyone involved about the status... on a new feature. We start with a story that describes the feature and what the criteria is for accepting...our users, we are able to make informed software design choices, resulting in software that helps our users...

...promotes scientific sharing The EDGE Program is designed to expand the number of systems that can be studied...genotype to phenotype studies New organismal systems described at this meeting were extraordinarily diverse. ...generating interesting drug-like compounds especially cardenolides which have been used to treat heart disease....disease. Caterpillars that eat milkweed generate cardenolides that tastes yucky (and is poisonous) for bird ...control who a female wants to mate with and if she decides to raise their offspring or leave them to their...of basic evolution of fungi and eukaryotes and includes the fungus that is killing frogs all over the ...

... that can help with the transition.” The lab-to-desk-job culture shock is a real thing. I experienced...challenging than lab work at times.” Sometimes a desk job is defined as sitting down at a computer and...consider as you make this transition. Of course, a “desk job” could entail any number of career paths--industry...colleague Mike Lemieux told me, “I find that at a desk, it is much easier to stick to timelines. This is...from legal, Diana in finance, and Wu in graphic design. You could dread these interactions, but you could... move away from calling non-academic careers and desk jobs “alternative careers.” Realistically, based...