Skip to main content
Addgene

Enter a sequence to perform a BLAST-based search of the Addgene plasmid sequence database.

Information

In Category

We narrowed to 849 results for: HA

Showing: 241 - 280 of 849 results

  1. Viral Vectors 101: An Introduction to AAV

    Type
    Blog Post
    ... and serotype, respectively. For example, AAV2/8 has a type 2 ITR and a type 8 serotype. Almost all vectors... components      Cargo capacity Empty rAAV has a cargo capacity of ~ 4.7 kb, meaning that your gene...usually quite stable and in some mammalian tissues has shown expression for several years (Wonjo et al.,...scAAV vector with half the size of the ssAAV genome has a mutation in the terminal resolution site (TRS) ...

  2. Mapping the 4D nucleome with CRISPR/Cas9

    Type
    Blog Post
    ...nuclease activity, catalytically dead Cas9 (dCas9) has many applications, the most well-known being transcriptional...transcriptional activation/repression. dCas9 has also previously been adapted to fluorescently label a sequence...imaging time for this technique. The NIH Common Fund has made mapping the 4D nucleome a specific priority,...

  3. New Videos: Addgene Lab Tips

    Type
    Blog Post
    ...Addgene has distributed over 800,000 plasmids since 2004. In that time, our lab has gotten a lot experience...

  4. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    ... population level, but each individual cell only has a maximum of one gene knocked out. In this genetically...containing up to around 200,000 different gRNAs. It has been suggested that researchers should use ~400X ...than gRNAs in a screen, to ensure that each gRNA has a good chance to contribute to the screen. In the... If you are lucky and find that a druggable gene has a central role in your system, you could majorly ...guest blogger Fredrik Wermeling! Fredrik Wermeling has a research group at the Centrum for Molecular Medicine...Medicine, Solna, Karolinska Institutet, in Sweden. He has a particular interest in using molecular tools to...

  5. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    ...promoter of the 5’LTR has been deleted to reduce its activity. A CMV or an EF1α promoter has been inserted in...The risk of generating RCL is low but extra care has to be taken especially if you are working with proto-oncogenes...previously used to drive expression from the LTRs) has been removed, Rev (which facilitates nuclear export...expression. In contrast, in the Tet-on system, tTA has been modified (and renamed rtTA) to bind TRE only...viral binding. This method, called spinoculation, has been found very effective for T cells transduction...

  6. RNA Interference in Plant Biology: New Tools for an Old Favorite

    Type
    Blog Post
    ...It is fair to say that CRISPR has revolutionized molecular biology and has enabled functional studies in...enzyme named Dicer (Figure 1). The plant kingdom has expanded its repertoire of Dicer enzymes and these...knockout methods of the same gene. However, recent work has called this prevalent assumption into question. Specifically...multiple target sequences are concatenated. hpRNA has been extensively used in plant research and many ...constructs are available to the community. Each construct has unique features, such as inducible expression of ...

  7. A History of Genome Engineering in Popular Culture

    Type
    Blog Post
    ... techniques is a speculation of yesteryear which has been realized recently. Science Fiction (SciFi) writers...even before the discovery of genes themselves.  How has scientific progress in genetic editing impacted the...technology on various non-scientific platforms, CRISPR has also transcended into the world of SciFi. This is...and most important genres of literary fiction. It has always served as a vision of the future and to make...and agriculture, its portrayal in popular fiction has been largely negative. But these negative depictions....1126/science.1258096 Gaudelli NM, Komor AC, Rees HA, Packer MS, Badran AH, Bryson DI, Liu DR (2018) Publisher...

  8. Finding Your Perfect Job After University

    Type
    Blog Post
    ...industry just wasn't for me, and as has been highlighted recently it has become more difficult to attain ...interesting and luckily genetics is a sector which has been rapidly growing. These factors left me optimistic...because it is a small and dynamic company and so it has the flexibility of a ‘start-up’. I was surprised ...schedule.  I really love my highly varied role, it has allowed me to travel and to share my passion for ...everyone. My career path may seem quite unusual but it has allowed me to try out working in many different sectors...

  9. Lab Automation at Addgene

    Type
    Blog Post
    ...Automating bacterial transformation One protocol that has been automated is high throughput transformation....benefits of the Biomek FxP is it’s customizability. It has a large deck that can house many different types ... how long to keep it there. In addition, the FxP has two robotic, mobile pods - one pod can pick up 96...cap sealer. This is a simple table top device that has a roller to go over caps and push them into micronic...

  10. Corralling and Cornering the Goldilocks Antibody

    Type
    Blog Post
    ..., that informs us that this antibody has specific binding and has a strong signal. We call the unique ...strong signal has a shorter exposure time than the negative control, and a weak signal has a longer exposure...

  11. Antibodies 101: Conventional vs Spectral Flow Cytometry

    Type
    Blog Post
    ..., full spectrum flow cytometry (“spectral flow”) has emerged as a particularly useful tool for those wishing...for PE. In order to correct for this, compensation has to be applied to “subtract” this spillover and correctly...they can more easily be detected. Cytek’s website has a convenient tool, called the similarity index, which...beads or cells. Overall, spectral flow cytometry has the ability to generate incredible amounts of data...

  12. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    ...   Fig. 1: SpCas9 has sparse PAM coverage in the human genome. SpRY Cas9 has PAM sites covering the... a PAM with no sequence constraints, or at least has looser or different requirements. Luckily, there ...alternative PAM requirements (Zhao et. al, 2023). xCas9 has been shown to be less effective at NGH PAMs, though...

  13. Adeno Associated Virus (AAV) for Cell and Gene Therapy

    Type
    Blog Post
    ... development and function. To date, gene therapy has been used to treat 10 infants with this disease. ... nucleic acid cargo (DNA or RNA) to target cells has been challenging. A variety of vector delivery technologies...AAV is not associated with any pathogenicity and has a low immunogenic profile. AAV was first discovered...Harshana S De Silva Feelixge is a researcher whose work has focused on gene therapy for viral infections. She...

  14. Plasmids 101: Terminators and PolyA signals

    Type
    Blog Post
    ... transcriptional read-through. Chris Voigt’s lab has characterized a set of prokaryotic terminators and...for eukaryotic termination are understood and it has been noted that each eukaryotic RNA polymerase terminates...combination with certain viral packaging systems has been associated with reduced viral titer, but improved...Expression Levels of Lentivirus-Delivered Transgenes but Has the Potential to Reduce Viral Titer in a Promoter-Dependent...

  15. CRISPR 101: Mammalian Expression Systems and Delivery Methods

    Type
    Blog Post
    ...to see if anyone has published a protocol that would work for your system. Addgene has depositor submitted...Sep 30, 2020 by Marcy Patrick. CRISPR technology has been widely adopted for genome editing purposes because...Mammalian CRISPR expression systems Each model system has its own best practices for efficient delivery of ...

  16. Story of a SynBio Startup: RevBio's Epiphany (or Lack Thereof)

    Type
    Blog Post
    ...understands (synthetic biology), building a product no one has ever seen before (flowers that change color throughout...but the experience is invaluable. This opportunity has given us the chance to develop the technology we ...the business training to make the most of it. It has also allowed us to really analyze what we want out...Nikolai Braun grew up outside of Washington D.C. and has always enjoyed exploring the outdoors and experiencing...

  18. CRISPR Challenges: Standardization and Homology Directed Repair

    Type
    Blog Post
    ...KSgenome and seeing the intricacies of DNA repair has emphasized how essential these studies are to improving...alternative non-homologous end joining (Alt-NHEJ), has been harnessed for CRISPR knock-in applications. ...29266098 Creating knock-in mouse models with CRISPR has been challenging, with protocols using dsDNA templates...

  19. CUT&RUN: An Improved Method for Studying Protein-DNA Interactions

    Type
    Blog Post
    ...for mapping where a protein binds genome-wide, and has been widely applied in many model organisms and cell...proteins (Figure 1). Importantly, since Protein A has weaker affinity for mouse IgG, it is helpful to incubate...prior to the addition of Protein A MNase. Once MNase has been localized to target sites, the nuclease is briefly...

  20. The Strength of Story Telling

    Type
    Blog Post
    ...Show that as the story has progressed, you or your lab or science or your CV has grown as well. Find your...Task, Action, Result) interview technique, which has been addressed in much better pieces than I’m able...

  21. Sensing Neuronal Dopamine

    Type
    Blog Post
    ...measuring DA and other neurotransmitters in the brain has been in vivo microdialysis of extracellular fluid... dLight1 dopamine sensor In 2018, Lin Tian’s lab has developed dLight1, a molecular tool to detect and...here! The GRABDA dopamine sensor Yulong Li’s lab has created the GRABDA sensor based on a similar strategy...

  22. New and Upcoming Viral Vectors - Spring 2019

    Type
    Blog Post
    ...AAV5  pOTTC1596 - pAAV SYN1 HA-hM3D(Gq) 121538  AAV5  pOTTC1484 - pAAV SYN1 HA-hM4D(Gi) 119741  AAV5 ...iGABASnFR GABA sensor (pAAV.hSyn-FLEX.iGABASnFR) has good membrane localization and brightness. The F102G...

  23. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    ...require heavy optimization. Claudia Vickers’s lab has designed novel SL fluorescent biosensors by integrating...showed that the receptor-based fluorescent biosensor has a high sensitivity (50 nM-500nM range) and a high...recent months: Our selection of GCaMP8 AAV preps has expanded. Check out the new selection of ready-to-use...

  24. Year of the Camelid: Antibody Style

    Type
    Blog Post
    ...The UN General Assembly has declared 2024 the International Year of Camelids. The declaration is intended... antibody libraries to generate new sdAbs. There has also been some interesting work done developing transgenic...al., 2021.   The single-domain antibody field has come a long way since the serendipitous discovery...

  25. Hot Plasmids - October 2022

    Type
    Blog Post
    ...channelrhodopsins. HcKCR1 has a maximum absorption of 540 nm and two-photon excitation around 1,040 nm, has larger photocurrents...anti-c-Myc [9E10] (Even, et al., 1985), the Myc tag has become one of the most commonly used protein tags...

  26. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ...inhibitory GABA-ergic interneurons, Gordon Fishell’s lab has developed a new AAV approach using regulatory elements...fluorescent protein - mScarlet! The Dorus Gadella lab has recently developed a synthetic red fluorescent protein... imaging and in protein fusions. The Gadella lab has deposited all mScarlet constructs with Addgene. ...efficiency Blue flame depositor Yannick Doyon’s lab has developed a new method to increase CRISPR editing...increases the probability that the locus of interest has also been edited! Thus, ouabain selection enriches...coselection are available at Addgene, and the Doyon lab has also provided a detailed protocol at Protocol Exchange...Essential gene Knockdown library. Although B. subtilis has a fully sequenced genome, many applications in biotech...

  28. Top Requested Plasmid of 2017 - pMD2.G

    Type
    Blog Post
    ...! Lentivirus envelope vector pMD2.G This plasmid has been requested nearly 6,000 times (over 800 orders...broad usefulness of pMD2.G and VSV-G. This plasmid has been used in studies of the secretome, ubiquitin ...

  29. Plasmids 101: Restriction Cloning

    Type
    Blog Post
    ...and insert a linear fragment of DNA (insert) that has been cut by compatible restriction enzymes. An enzyme...one found on the left side of the image below. It has a promoter (blue arrow) followed by the restriction...finished plasmid   Once it looks like your ligation has worked, you will need to pick individual bacterial...purpose. Ideally, once you know that your plasmid has an appropriately sized insert, you should send it...plasmid for more details. Once your complete plasmid has been verified, you’re ready to get experimenting!...

  30. PCR Applications for Copy Number Variation Assays

    Type
    Blog Post
    ... ddPCR.) qPCR Quantitative/real-time PCR (qPCR) has been used for copy number assays for years. qPCR ...and quencher to quantify specific amplicons.  Sybr has high background noise, due to the large amount of...AT-rich DNA and the length of the amplicon and it has a higher rate of false positives (Colborn, 2008; ... used to accurately call copy numbers, though it has been shown to identify trends correctly (Muhamad,...

  31. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    ... disrupting normal brain function. Chemogenetics has become a popular tool in the field as it provides...chemogenetics are instead that it is noninvasive and has longer-lasting effects. Building your chemogenetic... agonist for muscarinic receptor derived DREADDs has been clozapine-N-oxide (CNO). However, concerns have...are already approved for clinical use. Varenicline has been further modified to create ultrapotent PSEMS...

  32. Plasmids 101: Degron Tags

    Type
    Blog Post
    ...In the last decade, the use of degron tags has become increasingly popular for the modulation of endogenously...protein levels can return to normal once the drug has been removed. The degradation and reversal are rapid... its first use as a protein regulation tool, AID has gone through an additional round of evolution (AID...the system, but the general concept of the system has remained the same. An obvious drawback of the system...

  33. Hot Plasmids February 2024

    Type
    Blog Post
    ...FIQ or mStayGold (Ando et al., 2023). This variant has similar photostability and brightness compared to... of AAV9-X1.1 to that of AAV1. This viral vector has brain endothelial cell-specific tropism, with slightly...purify or image a recombinant protein, the FLAG tag has proven itself a good option for essentially all applications...western blots, though the original parent clone (M2) has been widely used in a variety of applications. Find...

  34. Academic vs. Industry Postdocs

    Type
    Blog Post
    ...Not the case, as I’ve discovered.) As someone who has now seen both sides of the postdoc experience, I’... lab is closely involved with clinical trials or has ties to industry, the drug discovery process can ...Genentech, will not. The point is that each company has a unique approach, and some might suit your needs...currently a Postdoctoral Scientist at AstraZeneca. She has a background in cell and developmental biology and...

  35. Fluorescent Tagging of Endogenous Genes with SapTrap

    Type
    Blog Post
    ...the growing spectrum of fluorescent proteins (FPs) has been an invaluable resource for studying the organization...sequence (like GFP) can be inserted at the DSB. CRISPR has allowed scientists to tag and light up endogenous..., and T2A-TurboGFP-PEST), and small epitope tags (HA, Myc, Strep tag II, AviTag, HaloTag, SpyTag). The...characteristics. For example, Masato Kanemaki's lab has developed a CRISPR/Cas-based system for tagging endogenous...

  36. Addgene's Cancer Collection Pages Connect the Research Community

    Type
    Blog Post
    ...community has access to all of the latest resources? In response to this challenge, Addgene has launched... why recent work in the field of cancer research has focused on picking apart the complex cellular pathways...at Vanderbilt University in Nashville, Tennessee has already demonstrated that a number of novel small...

  37. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    ...host cells immediately prior to their transduction has been shown to increase AAV transduction efficiency... such cationic reagent that is commonly used and has been shown to increase transduction efficiency by...single-use aliquots. Never use a polybrene solution that has been thawed more than once. Fibronectin is another...polybrene (e.g., hematopoietic or primary cells), and has been shown to increase transduction efficiency by...

  38. Cpf1 Update: Comparison to Cas9 and NgAgo

    Type
    Blog Post
    ...regulatory elements. To solve this problem, the Zhang lab has previously used SaCas9 (1053 aa/3.2 kb) with AAV ... these options (2.7 kb with 24 nt guides), so it has the potential to eclipse any of the CRISPR nucleases...THE ORIGINAL NgAgo ARTICLE DISCUSSED IN THIS POST HAS BEEN RETRACTED AND FOLLOW UP STUDIES HAVE FAILED ...be the best option for HDR, but this possibility has not been experimentally confirmed. Researchers have...

  39. Plasmids 101: Colony PCR

    Type
    Blog Post
    ...for the presence of an insert. Traditionally this has been done with restriction enzyme digest; however... Find Empty Backbones at Addgene Each approach has advantages and drawbacks, which are summarized in...insert-specific primers but also tell you whether the insert has the correct directionality. Verifying the insert ...and are more likely to work in a PCR reaction that has bacterial debris. Use a positive control. A good...

  40. Which Fluorescent Protein Should I Use?

    Type
    Blog Post
    ...Fluorescence is the emission of light by a substance that has absorbed light. The emitted light is at a longer ...protein Excitation & Emission (ex/em): Each FP has its unique ex/em peak. Therefore, choose FPs that...detect the emission. For example, if your microscope has only two lasers, at 488nm and 561nm, you will not...Some proteins are very dim (e.g. TagRFP657, which has a brightness of 0.1) and this should be taken into...
Showing: 241 - 280 of 849 results