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Showing: 181 - 200 of 877 results
  1. New Tools Enable CRISPRa for Neuroscience Applications

    Type
    Blog Post
    Published
    July 2, 2019, 12:51 p.m.
    ...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 ...
  2. AAV Titers: Where Do They Come From And What Do They Mean?

    Type
    Blog Post
    Published
    Nov. 15, 2017, 3 p.m.
    ...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 ...
  3. Transferable Skills Guide: Career Planning Resources

    Type
    Blog Post
    Published
    Nov. 14, 2017, 2 p.m.
    ...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 ...
  4. CRISPR Activation: A Practical Guide

    Type
    Blog Post
    Published
    Aug. 18, 2016, 2:30 p.m.
    ...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...
  5. Single-cell tracking of lineage and identity with CellTag

    Type
    Blog Post
    Published
    May 9, 2019, 1:15 p.m.
    ...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...
  6. 10 Great Guest Posts We're Thankful For

    Type
    Blog Post
    Published
    Nov. 18, 2016, 3 p.m.
    ...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...
  7. Fluorescent Proteins 101: Photoactivatable Fluorescent Proteins

    Type
    Blog Post
    Published
    April 25, 2017, 2:30 p.m.
    ...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...
  8. The Many Reasons for Irreproducible Research- and a Vaccine to Eradicate It

    Type
    Blog Post
    Published
    Jan. 30, 2020, 2:15 p.m.
    ...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?...
  9. GPCRs: How Do They Work and How Do We Study Them?

    Type
    Blog Post
    Published
    Feb. 11, 2021, 2:15 p.m.
    ...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...
  10. From Lab Bench to Leadership: Transitioning into Management as a PhD

    Type
    Blog Post
    Published
    Jan. 21, 2025, 2:15 p.m.
    ... 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 ...
  11. Back to Bacteria: CRISPR gRNA Multiplexing Using tRNAs

    Type
    Blog Post
    Published
    June 2, 2015, 6:06 p.m.
    ...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 ...
  12. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    Published
    Feb. 27, 2024, 2:15 p.m.
    ... inserted into it. You will have the option to indicate separate promoters, tags, or cloning information...strain(s) through Addgene. Description – please indicate the gene cloned into your plasmid, any tags or...information. Enter plasmid information During the remainder of the process, you will enter plasmid data in...this will make it easier for other scientists to find information related to your plasmid. It is optional...insert using standard mutation notation. Please indicate the amino acid change as well as any functional...etc) on your insert. Use the drop-down menu to indicate whether the tag is on the N or C terminus and ...information will make it easier for scientists to find your plasmid by type. Figure 8: Add vector backbone...
  13. dTAG - You're it!

    Type
    Blog Post
    Published
    June 21, 2018, 2:06 p.m.
    ...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...
  14. Antibodies 101: Selecting the Right Antibody

    Type
    Blog Post
    Published
    March 15, 2022, 1:15 p.m.
    ...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...
  15. To Each HIS Own

    Type
    Blog Post
    Published
    March 2, 2023, 2:15 p.m.
    ...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 ...
  16. Is this the right place for me? 8 tactics for choosing a lab

    Type
    Blog Post
    Published
    Oct. 2, 2018, 12:56 p.m.
    ...options critically. How will it feel when you find it? If you find the right lab, some days it won’t even feel...scientists seem to toss data-driven reasoning out the window when making decisions. We think scientists make...success. Gather data on the lab choices How do you find out what the lab is really like? Gather data of ...boss You sign on to a lab for the mentoring only to find out that the lab head is an absentee advisor. They...character traits and skills that you hope to grow.  Find mentors who take their responsibility to train you...meaningless tasks, removal of responsibility, repeated reminders of blunders, setting target up to fail, shifting...(topic and/or technical aspects) applicable to industry if this path interests you  (e.g. human disease...
  17. Hot Plasmids: Spring 2025

    Type
    Blog Post
    Published
    June 3, 2025, 1:15 p.m.
    ... articles. Here's what you'll find in this post: Glutamate indicators with increased sensitivity and...viral preps targeting interneurons Glutamate indicators with increased sensitivity and tailored deactivation... for a variety of synaptic imaging applications. Find iGluSnFR4 plasmids here! Aggarwal, A., Negrean,..., Iyer, R., … Podgorski, K. (2025). Glutamate indicators with increased sensitivity and tailored deactivation...and AAV plasmid sets launching later this year!  Find Addgene's own Lentiviral backbone plasmids here!... 5% and 11%. It has higher efficiency and lower indel formation than methods relying on homology-directed...scarless, unlike previous integrase-based approaches. Find STITCHR plasmids here! Fell, C. W., Villiger, L...
  18. Split-BioID: An Improved Method for Studying Protein-Protein Interactions

    Type
    Blog Post
    Published
    May 1, 2018, 1:57 p.m.
    ...at Addgene.org Find plasmids from the Bethune Lab Find Plasmids from the Roux Lab Find Plasmids for FRET... particular protein  complex as opposed to an individual protein. The Bethune Lab has overcome this limitation...directly interact with the bait from those that indirectly interact or from proteins that are just in the...labeling. BioID requires 6-24 hours for tetracycline induced expression of the BirA* fusion protein and saturation...Schopp et al allowed 24 hours for tetracycline induced expression and biotinylation of interacting proteins... PMID: 28585547. PubMed Central PMCID: 5467174. Find the Split-BioID Plasmids Here! Munter, S.D., Görnemann...fragmented at a different site than in Schopp et al. Find these Split-BioID Plasmids Here!   Burke, B., Kim...
  19. Hot Plasmids - October 2020

    Type
    Blog Post
    Published
    Oct. 13, 2020, 1:15 p.m.
    ...deposits and we hope they'll make it easier for you to find and use the plasmids you need. If you'd ever like...deposit please sign up here.  Here's what you'll find in this post: Tools for labeling excitatory inhibitory...respectively. Once expressed in the neuron, the FingRs bind to the endogenous synaptic proteins allowing them...inhibitory or excitatory synapses or that are Cre-inducible, researchers can explore synaptic integration ... show a high proportion of blue fluorescence, indicating cell proliferation. BaF3 cells maintained in ...in high IL-3 showed mostly blue fluorescence, indicating a quick cell cycle. Cells that had IL-3 withdrawn...a few highlights from recent CRISPR plasmids. To find all of the CRISPR plasmids available from Addgene...
  20. What's New in CRISPR - May 2019

    Type
    Blog Post
    Published
    May 28, 2019, 12:49 p.m.
    ...Addgene.org Read our CRISPR guide Find CRISPR plasmids by function Find recently deposited plasmids ... but we hope that this blog series will help you find new CRISPR tools for your research! This time: ...control. Read the paper in Nature Communications Find the E. coli genome-wide CRISPR inhibition library...fold. Read the paper in Communications Biology Find the Cas9-PCV and PCV-Cas9 plasmids Selective curbing... editing efficiency. Read the paper in Nature Find the SECURE base editors All-in-one AAV delivery... in zygotes. Read the paper in Molecular Cell Find the Nme2Cas9 All-in-One AAV constructs CRISPR interference...essential C. albicans gene. Read the paper in mSphere Find the dCas9 and dCas9-Mxi1 plasmids If you have a...
Showing: 181 - 200 of 877 results