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...used to infer differentiation trajectories and identify both up- and down-regulated pathways. Individual...unique barcodes for each TF help streamline identification. While the Zhang lab controlled their multiplicity... a model system. TF screens can be designed to identify custom TF programs to acutely initiate disease...

...CRISPR locus of a bacteria host encodes multiple guide RNAs (gRNAs) on a single array to target the genome...applications when repetitive tRNA sequences must be avoided, different tRNAs with optimized 5’ leader sequences...array for multiplex gene editing applications, we provide detailed manual and online design examples of using...

...coinjection into zygotes of Cas9 mRNA and a single-guide RNA targeting the mutant allele (7). An independent...CRISPR/Cas9 has been used to generate mutations in a wide range of organisms, but its potential to efficiently...These papers suggest it might work well enough to consider as a therapeutic.” There are many fundamental ...

...the pMinDisplay backbone is fused to a specific peptide segment (PDGFR TMD) that anchors the sensor to ...the membrane and displays it on the extracellular side. In addition to the original PDGFR TMD, the researchers...ordered directly from Addgene. The Podgorski Lab has provided a brief breakdown of the promoter systems you ...

...that interact with the virus. As a result, we identified 332 human proteins that the virus interacts with...the help of human proteins in their lifecycle, identifying these potential human helpers of SARS-CoV-2 (...You could be the next scientist joining in this worldwide fight against SARS-CoV-2. As we’ve all been affected...

...bioscience and tech transfer communities who help guide and promote Addgene in its mission to make research...given by Phil Sharp. After a few rotations, Doench decided to complete his PhD work in Sharp’s lab.  Doench...a tremendous amount of technology development alongside it.” Doench starting working with David Root ...

...proteins to be imaged together, you will need to consider the same factors as when choosing an individual... the filter sets or lasers on your microscope. Ideally, you would like to have substantial overlap between...protein and a filter set, many filter set vendors provide tools to plot the fluorescence spectra of proteins...

...based on maps of protein structure. Sometimes this provides a great deal of clarity. But, often these methods...Institute Target Accelerator Plasmid Collections, now widely available to the scientific community through Addgene...Functional Interrogation of Rare Cancer Variants Identifies Oncogenic Alleles. Kim E, Ilic N, Shrestha Y....

...check out this blog post or see Addgene's CRISPR guide page for a review of Cas9.  For a brief comparison...Essletzbichler, P.,  Zhang, F. (2015). Cpf1 Is a Single RNA-Guided Endonuclease of a Class 2 CRISPR-Cas System. Cell...Research Catch up on Your CRISPR Background with Our Guide Pages ...

...through our hot plasmids articles. These articles provide brief summaries of recent plasmid deposits and ... the first reporter genes using ultrasound and provide a major benefit to scientists studying microbiome...different cells present in a mixed population, or identify differentiation and disease states. This collection...

...there are many great resources available to help guide you through the rough waters of mutation validation...great deal of precision. This means digital PCR provides an absolute measure of target abundance, does ...screening to validate your genome edits, this paper provides detailed instructions on how to design such assays...

...promoter should flank this component as well. On both sides of this promoter-gene combo are two long terminal... and have appropriate factors displayed on the outside of the particle to interact with the host. This...applications and aid in ease of use. Our lentiviral guide outlines these components.     Fig. 1 - ...

...within a biological system. Basically, biosensors provide quantifiable readouts for levels of different chemicals...FRET-based biosensors have been engineered to detect a wide array of neurotransmitters ranging from dopamine...fluorescence-based and have been optimized for a wide-range of uses to monitor the fundamental cellular...

... kB, then you would be oversized even before considering the need for some manner of polyA. In this scenario...can be safely concentrated to ~1x1014 vg/ml.. As evident in our repository, this application is an outlier...Addgene.org Find Adenovirus Plasmids Adenovirus Guide Page Subscribe to Viral Vectors Blog Posts ...

... and gotten unusual or unexpected results? Or considered why DpnI will degrade your template DNA from ...methylated (if it comes from a phage for instance). Aside from restriction modification systems, DNA methylation...level, the potential for methylation should be considered when digesting DNA. Why? Well, even though Dam...

...task and will prevent you from getting lost in the wide word of informatics. I am a molecular biologist ... project a few years ago when my advisor and I decided to do a massive RNA-seq analysis. I had never interacted...even if your conclusion is “this approach is not ideal because…” Including a conclusion for each analysis...

...through our hot plasmids articles. These articles provide brief summaries of recent plasmid deposits and ...can be reconstituted in vivo to form a protein identical in sequence to the unmodified enzyme. Each half...any exogenous sequences and regenerates a native peptide bond at the split site. Figure 1: The base...

...interactome of these key regulatory molecules, and has provided clues to their previously uncharacterized cellular...acetyltransferase to luciferase, the basic assay remains widely used today.  The sheer abundance of these critical...greatly increased the burden associated with identifying plasmids in the research literature and acquiring...

...molecular tools to map neuronal connectivity are widely used in neuroscience. In this post, I’ll describe... in first order neurons, the G-deleted RABV can divide and replicate (since these processes are mediated...Another study used RABV monosynaptic tracing to identify inputs to two major dopaminergic cell populations...

...specificity of AAV, work has focused on modifying the outside protein shell, or capsid, of the virus so that ...here.  Applications for synthetic promoter AAVs Besides restricting AAV transgene expression to particular...synthetic promoters with high rates of labeling are ideal of gene therapy applications, promoters that produce...

...initiation step of gene transcription; today we'll provide an overview on how transcription stops, or termination...post-transcriptional additional of multiple adenine (A) nucleotides to the tail of a messenger RNA transcript. The...be detailed here, but additional references are provided at the end.  Nearly all common bacterial expression...

...SpCas9. Hu et al. performed GUIDE-seq analysis to characterize genome-wide off-targets, since increasing... Resources on Addgene.org Check out our CRISPR Guide Find CRISPR Plasmids for Mammalian Systems Learn...

...Even before fluorescent proteins (FPs) came into wide use, there were a variety of ways to monitor cell...few properties that have now become hallmarks of ideal FP timers: 1. Ratiometric determination of expression...Empty Backbones Check Out Our Fluorescent Protein Guide ...

...that popularity through the years.  Libraries are divided into different categories based on function: knockout... years, even as more CRISPR technologies become widely used and available. If we look closely at Figure...navigate that learning curve. As of 2023, our CRISPR guide has been viewed 1,904,462 times, and the CRISPR ...

...Degrons are destabilizing domains which will be identified by various enzymes and sent to the proteasome...protein for imaging. Given its popularity and widespread usage, a proteolysis-targeting chimera (PROTAC...some stage, then choosing a HaloTag could pay dividends down the road. It is difficult to predict precisely...

...below. Types of Voltage Indicators GEVIs can be divided up structurally into 2 main groups: voltage sensitive...voltage sensing component (i.e., rhodopsin naturally resides in the membrane), or both. Targeting these systems...Addgene’s Virus Protocols Addgene’s Chemogenetics Guide Resources on the Addgene blog Viral Vectors ...

...to not produce artifacts in your experiments. Considerable effort has gone into making FPs monomeric, yet..., increasing the probability that two FPs will collide with each other. The consequences can be dramatic...localized to the endoplasmic reticulum, the normally spiderweb-like pattern of tubules can be grossly distorted...

...factor of 2. This means that if you set up two identical assays with the same sample, you could end up ... should fall within the range of the assay for a wide range of titers. We usually dilute our samples 1...experiments.  References Furuta-Hanawa, Birei, Teruhide Yamaguchi, and Eriko Uchida. "2D droplet digital...

...sites. FRT sites and loxP sites differ at the nucleotide level but share an overall structure of 13 base...37 °C). Smart molecular evolution led to the identification of FLPe which has a temperature optimum of ...Recombination between the two target sites with identical sequence leads to inversion of the flanked DNA...

... codon, stop codons, reading frame considerations, etc). This video demonstrates how to use the Snapgene...Protocols Watch Our Diagnostic Restriction Digest Video Check Out Our Molecular Biology Reference Pages  ...

...more funding and research opportunities are being provided to immunotherapy than ever before. Although different...from many other types of cancer. Mesothelioma is considered an orphan cancer; it is rare disease with only...(ADAO) are working toward such a ban, and the president of the organization, Linda Reinstein – whose husband...

...results in a polymerase error rate of up to 2% per nucleotide position. The “sloppy” PCR reaction is created...ssDNA is packaged into phage and then extracted. Besides taking a couple days to generate the phage-derived...strand: A mix of degenerate primers and the high-fidelity Phusion polymerase are used to introduce point...

...fluorescence can occur. Cre recombinase can be used alongside common, existing tools for fly research that use...turnover of Langerhans cells in the dermis and identify that Langerhans cells are replenished through ...The intestinal stem cell markers Bmi1 and Lgr5 identify two functionally distinct populations." Proceedings...

...different, so more options mean better chances of identifying a successful editor. For this application, ABE8e...And base editing advances won’t stop here. By coincidence, KJ’s genetic sequence was able to tolerate bystander...Read our CRISPR 101 eBook Brush up with our CRISPR Guide ...

...found the video radd-ically helpful!    More resources at Addgene Addgene's Antibody Guide What is an...

...Innovation (IPI). Together, we’ll be working to provide researchers with a collection of IPI’s well-validated...on our website and blog, including our antibody guide, antibody protocols, and Antibodies 101 blog series...

...      And you’ll be able to find all published videos in our Quick Tips playlist:  Want your quick tips...be sharing our favorite parts of every quick tip video on our TikTok! Be sure to subscribe to our YouTube...

...your favorite name. We hope you submit your best ideas because the finalists will receive some newly designed...Learn more about Blugene's make-over Get a how-to guide on the new Addgene search engine ...

...Sorting” is there to help guide you through the process. If you’re confident in your application and protocols...

...Learn more about the Data Hub in this blog post. Alongside our efforts to facilitate the sharing of reagents...Learn more about the viral service Read our AAV guide ...