We narrowed to 890 results for: IND

...of making targeted genetic edits. Cas9 is a DNA binding protein that can be directed to virtually any genetic..., each of which cleaves one strand of DNA upon binding the protospacer. The resulting double strand break...respectively. When complexed with a truncated gRNA, Cas9 binds the target site, but does not cut either strand ...fusion proteins retain the ability to recognize and bind to DNA; when complexed with a gRNA that directs ...of attempting to simultaneously and selectively induce both cutting and regulation of expression at different...of complementarity to a target site, Cas9’s DNA binding capability remains intact while its nucleolytic...transcriptional start site (TSS) of a gene of interest, it induces potent gene expression with no genetic alterations...

...either the N or C terminal of a protein and can inducibly facilitate the degradation of the tagged protein...mechanisms of control. Protein degradation can be induced by the addition of a small molecule, and protein...degron tags can do more than just degrade! Read on to find out more. Common degron tags When it comes to degrons...novel degrader, dTAG13, is a small molecule which induces dimerization of the FKPB12F36V fusion protein with...also be handy!   Fig 1. dTag molecules induce dimerization of an FKBP12F36V fused target protein-of-interest... can be added to media, buffer, etc., which will bind and light up a tagged protein for imaging. Given...ligases to specific proteins by simultaneously binding both their target protein and an E3 ligase through...

...Learn about Addgene's viral service Find viral protocols and videos Find biosensor AAVs for your research...at a certain frequency, and this frequency can indicate behavioral state, for example, if someone is awake...) are well understood, the direct effect of DA binding to a target neuron is more mysterious. Thus, scientists... ms on and 100 ms off), enabling detection of individual peaks of DA release.  Using dLight1 sensors in...during learning and reward consumption in live mice Find dLight sensor here! The GRABDA dopamine sensor Yulong... such that the GFP fluoresces upon DA-receptor binding (Sun et al., 2018). These sensors also exhibit ... mice over the course of Pavlovian conditioning Find the GRABDA sensor here! Overall these sensors are...

...plasmid, scientists would spend hours to days trying find their correct clones. Additionally, the presence...vice versa). Looking around the lab, you'll likely find many of the antibiotics listed in the table below...Working Concentration** Kanamycin aminoglycoside Binds 30S ribosomal subunit; causes mis-translation Bactericidal...Bactericidal 50-100 ug/mL Spectinomycin aminoglycoside Binds 30S ribosomal subunit; interrupts protein synthesis...Bactericidal 100 ug/mL Bleomycin glycopeptide Induces DNA breaks Bactericidal 5-100 ug/mL Erythromycin...Bactericidal 10-100 ug/mL Tetracycline tetracyclin Binds 30S ribosomal subunit; inhibits protein synthesis... Bacteriostatic 10 ug/mL Chloramphenicol   Binds 50S ribosomal subunit; inhibits peptidyl translocation...

...on PHP.B binding in vitro. They found that when LY6A contained C57BL/6J SNPs, PHP.B could bind it, but ...an ELISA to measure binding of PHP.B to a soluble version of LY6A. They saw binding between PHP.B and the...Wilson lab at the University of Pennsylvania independently searched for this unique sequence of DNA and...susceptibility. The Deverman and Wilson Labs independently identified LY6A as a receptor for PHP.B by looking...search on SNPs and small insertions and deletions (indels) most likely to have large phenotypic effects such...but when LY6A contained BALB/cJ SNPs, binding was reduced. The Deverman group then looked to see if PHP....mutation in PHP.B’s unique capsid loop disrupted binding. Collectively these results suggest that PHP.B ...

...our comprehensive CRISPR guide Find CRISPR plasmids and resources Find CRISPRa plasmids ...engineered combinations of transcriptional activators to induce gene transcription at genomic sites targeted by...cDNA libraries. However, using CRISPRa to target individual genes can be relatively laborious because multiple... control over its expression. Constitutive or inducible Cre recombinases can be introduced by AAV or by...in defined substructures of the CNS as well as individual neuronal populations.  The Yang Lab’s AAV plasmids... promoter, which was superior in its ability to induce CRISPRa in primary rat neurons over other promoters...straightforward than generating cDNA for each gene isoform individually. To highlight this advantage, they selected ...

...are addressed with ddPCR technology (Hindson et al., 2011, Hindson et al., 2013), which has been shown ...2011): 8604-8610.4. Hindson CM, Chevillet JR, Briggs HA, Gallichotte EN, Ruf IK, Hindson BJ, Vessella RL,...Resources on Addgene.org Find All Available AAV Learn More about Virus Production Find Viral Vector Protocols...value, physical titers don’t necessarily give any indication of the infectious titer. The infectious titer...Although you can optimize your primers for each individual sample, this will reduce convenience and comparability... the assay. This may be due to competition for binding sites between the primers and the AAV genome repeats...Finally, because physical titer does not give an indication of infectious titer, consider titrating each ...

...employer in mind, you may consider starting with indeed.com or monster.com, where you’ll find a variety ...employment within industry or academia; although the ACS career fair is more industry-focused. For another...another industry-geared option, try the Biospace career events page. Biospace holds career events all over...institution’s career page, where you’re likely to find current job openings. A good approach is to do this...of academia, you may need to look a bit harder to find your mentor. This may be a connection you’ve made...and other popular job site webpages. Importantly, find a good mentor that can share relevant experience...www.gradschools.com/get-informed/grad-school-roadmap Find Job Opportunities for Scientists on docjobs.com ...

...example, CRISPRa can be used to find potential cancer drug targets by finding genes that cause cell death ...Used with AAV Resources on Addgene.org Find Cas9 Activators Find Cas9 Repressors Catch Up on Your CRISPR...CRISPR activation has been used to differentiate induced pluripotent stem cells (Chavez et al., 2015) into...Additionally, CRISPR-based activation can be made inducible upon environmental triggers. This is done by creating...activators, similar tools were made using other DNA binding proteins such as zinc fingers (ZF) and TAL effectors...by putting the promoter of the gene into a gRNA finder such as WU-CRISPR (Wong et al., 2015)) or our lab....2018.04.011 Polstein LR, Gersbach CA (2015) A light-inducible CRISPR-Cas9 system for control of endogenous gene...

...University in St. Louis provides a combinatorial indexing strategy that simultaneously tracks clonal history...variable region acts as a unique identifier for individual cells. Each cell is labeled with 3-4 tags to ...decreased over time but they were not silenced.    Find CellTag libraries here!  Applications of CellTag...reprogramming of mouse embryonic fibroblasts (MEFs) into induced endoderm progenitors (iEPs), a self-renewing cell...takeaways of CellTag CellTag is a combinatory cell-indexing tool that combines scRNA-seq and lineage tracking...identity changes, such as the differentiation of induced pluripotent stem cells or the clonal evolution ...cancer. If you’re looking to use CellTag, you can find the CellTag libraries at Addgene. The Morris Lab's...

...post are engineered to bind RNA sequences derived from Zika virus and, upon binding, produce reporter proteins...posts written by Addgene depositors. Below you’ll find brief summaries of several excellent guest posts... post, Luke Lavis from Janelia Research Campus reminds us that, although fluorescent proteins are incredibly...used in a wide variety of experiments. Read more to find chemical fluorophores for your next experiment. ...virus biology while leaving virulent components behind. Minigenomes are stripped down viral genomes containing...post for potential applications of R-bodies and to find a video of them in action. AAV: A Versatile Viral...Medical School describes how AAVs function and indicates important things to consider if you’re getting...

...Addgene.org Find Photoactivatable FPs in Empty Backbones in the Michael Davidson Collection Find Photoactivatable...from a dark state to a bright fluorescent state (“kindling”), and from a bright state to a dark non-fluorescent... In 2008, scientists used mutagenesis screens to find a version of Dronpa that doesn't become inactivated...photoactivation. A high contrast ratio usually indicates that a PA-FP has a low level of spontaneous fluorescence...photobleaching and phototoxicity that standard FPs might induce though their continuous excitation. By generating...photoswitching between two fluorescence wavelengths. Indeed, within the world of super resolution microscopy...localization information in cells. A point to keep in mind- although PA-FPs are vastly improved over the conventional...

...proteins that bind the RNA motif will be biotinylated. Read more about RapID in this blog! Find plasmids for...biotin. Biotin, is a natural coenzyme, that strongly binds the glycoprotein, avidin or a similar protein, streptavidin...biotin molecules to dissociate from the ligase and bind to exposed lysines on neighboring proteins. Thus...organoid or in vivo studies (Che and Khavari, 2017). Find APEX plasmids at Addgene! Since the development ...biotin ligase from A. aeolicus, which lacks the DNA-binding domain (233 amino acids) found in the E. coli version...fusion proteins to specific locations in the cell.   Find BioID2 plasmids here APEX2: A more sensitive ascorbate...Lam et al. 2014). They named this mutant APEX2.  Find the APEX2 plasmids! TurboID and minturbo: Rapid ...

...question after credible attempts to replicate major findings could not be replicated by other researchers.There...replicate.  Why is replicating previous research findings so difficult to do?  There are four reasons that... to present only the most novel or significant findings, and combined with powerful cognitive biases such... such as motivated reasoning and hindsight bias, we may fool ourselves and our readers into believing ...research practices (QRPs) can quickly lead us to find statistically significant results that are not robust... not come out the same as an original research finding we can be more certain that some important, previously...makes a big difference for the robustness of the findings.   How do we make research more reproducible?...

... PhDs Making the Leap to Industry." He successfully transitioned to industry after his PhD and a couple... PhDs face when transitioning from academia to industry is whether to stay hands-on in the lab or pivot...exploring alternative career trajectories within the industry, but the transition wasn’t easy - I had to reinvent...assistant. However, I recognized the need to shift my mindset to empower my team and allow myself to focus on... rather than the day-to-day details. Ignoring individual differencesManaging a team isn’t “one size fits...a diverse team where some members thrived on independence, while others required more guidance and structure...learned that my approach needed to adapt based on individual needs. For instance, I had one colleague who ...

...works GPCR signaling is initiated when a ligand binds to the extracellular surface of the GPCR. This results...from the Gβγ dimer. The Gα and Gβγ subunits then induce or inhibit intracellular signaling cascades as .... Ligand dissociation from the GPCR allows for binding of a new inactive heterotrimeric G protein complex...identify some of these unknown ligands and their GPCR binding partner, Bryan Roth’s lab created an open source...activation for more than 300 GPCRs. In this assay, binding of an agonist recruits a beta-arrestin-TEV protease... discover new ligands for unknown GPCRs. You can find the entire PRESTO-TANGO kit at Addgene! TRUPATH:...were first mutated via site-directed mutagenesis to bind non natural ligands. These non natural ligands however...

...100% indel frequency at both targets (35% and 76% biallelic), compared with only 44% and 66% indel frequency...recognition and cleavage. This sequence flexibility indicates that gRNAs could be incorporated into these constructs...the 5’ end. PTG construct schematics. Scissors indicate cleavage sites. After confirming gRNA cleavage...cleavage, they examined gRNA expression levels and indel generation for two PTG constructs in rice protoplasts... allowing them to examine the frequency of both indels at each locus, as well as large deletions. They...deletion with 4-20% frequency at the four loci, indicating that Cas9 can be guided to at least 8 distinct....1073/pnas.1420294112. Epub 2015 Mar 2. PubMed. Find the plasmids from this publication at Addgene ...

...those of antibody-independent assays such as mRNA data from transcriptomics; independent antibody strategies... internet search for the required antibodies and find….hundreds of options. With limited time and budget...immunoprecipitation, the epitope may not be accessible for binding and the antibody will not work. Whenever possible...more detailed data on the assay methods. You might find that while the website highlights a variation of...fluorophore, or a conjugated secondary antibody that will bind to your primary antibody. If you are trying to visualize...applications use either direct (primary-conjugated) or indirect (secondary) methods. In those cases, remember ...the antibody cross-reacts with. Ideally you will find an antibody that has been validated in the species...

...small molecules that bind these target proteins to other small molecules that bind an E3 ligase. These ...molecules by conjugating a CRBN-binding ligand (thalidomide) to an FKBP12F36V binding ligand (ortho-AP1867) via...E3 ligases such as cereblon (CRBN) or von Hippel-Lindau (VHL) to bring the endogenous degradation machinery...selectivity, overcoming resistance to inhibitors, and inducing prolonged biological effects. While targeted ...oncoproteins that lack selective small molecule binders. To overcome this limitation, we created a generalizable... FKBP12F36V-tagged proteins. The dTAG molecule induces ternary complex formation between the FKBP12F36V-fusion...and the ubiquitin proteasome machinery through binding with cereblon, leading to rapid target degradation...

...interest flowed through. The protein of interest binds to the metal ions, becoming immobilized itself. ...containing resin. The protein of interest (purple) binds to the Ni2+ and is immobilized along with a polyhistidine...purification, however, is the propensity for nonspecific binding of untagged proteins. Despite being a relatively...histidines are present in most production systems and can bind and co-elute with the protein of interest (Bornhorst...be high enough to disrupt contaminant-metal ion binding without eluting the target protein and needs to...substrates along with stringent wash conditions.   Find polyhistidine tag plasmids here! Where should I ...Unlike other affinity purification methods where binding relies on a specific protein conformation, the ...