Skip to main content
Addgene

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

In Category

We narrowed to 1,053 results for: FER

Showing: 391 - 420 of 1053 results

  1. CRISPR 101: Cytosine and Adenine Base Editors

    Type
    Blog Post
    ...updated by Jennifer Tsang in August 2020 and by Emily P. Bentley in September 2025. References and Resources...for resolving DNA double-strand breaks (DSBs) preferred by researchers. Compared to non-homologous end...APOBEC3G displayed less predictable sequence preferences.  The Liu lab also used natural and engineered...generation of base editors. Compared to BE3, BE4 offers a 2.3 fold decrease in C→G and C→A products as ...yielded an editor with more flexible sequence preferences, evoAPOBEC1-BE4max, which is significantly more...the development of prime editing, which uses a different mechanism than base editing, has allowed scientists...Resources References Chen, L., Zhu, B., Ru, G., Meng, H., Yan, Y., Hong, M., Zhang, D., Luan, C., Zhang,...

  2. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    ... the transcript. Different Cas orthologues and subtypes function somewhat differently and can be more ...Transcriptional repression (CRISPRi) in bacteria As RNA interference does not function in bacteria, most efforts ...promoters or direct gene knockout. In contrast, CRISPR offers a much more user-friendly way to modulate gene ...Peters, Oren Rosenberg, and Carol Gross used two different genetic systems, targeting either Gram-negative...bacterial species (Peters et al., 2019). The systems differ on how exactly the CRISPRi locus is introduced ...Clostridium difficile, could be targeted and preferentially depleted from gut. While still far from the...the BE3 that further improves editing rates. References Bikard D, Euler CW, Jiang W, Nussenzweig PM, ...

  3. CasPEDIA: A Functional Classification of Cas Enzymes

    Type
    Blog Post
    ...new features to the tool. Resources and References References Adler, B., Trinidad, M., Bellieny-Rabelo...solve the problem: CasPEDIA, a new resource from Jennifer Doudna’s lab at the Innovative Genomics Institute...CasID, to help facilitate direct comparison of different enzymes. CasPEDIA is meant to be a constantly ...plasmids containing that enzyme, and embedded in the References lists on every page you can find direct links...design, etc.). With this feature, you can compare different Cas enzymes to make sure you are getting the desired...the winner!).   Fig. 3 – Comparison of different Cas enzymes using CasPEDIA. Adapted from Adler...

  4. Viral Vectors 101: Inverted Terminal Repeats

    Type
    Blog Post
    ... 2019), and different combinations of ITR and capsid serotypes vary in their gene transfer efficiency....DNA molecule as the gene that they package. In a transfer plasmid, the transgene is flanked by two ITRs,...and rAAV vectors, modifications can affect gene transfer efficiency and transgene expression (Zhou et al...expression can be altered by using ITR sequences from different AAV serotypes that vary in their RBE and T-shaped...restriction digest methods.    Resources and references More resources on the Addgene blog Viral Vectors...Addgene.org Addgene's AVV guide AAV Viral Preps References  Chen, Y., Hu, S., Lee, W., Walsh, N., Iozza...

  5. New and Upcoming Viral Vectors - May 2020

    Type
    Blog Post
    ... various AAV serotypes can be used to achieve different patterns of expression, researchers often need...empirically for their own experiments. To this end, we offer control AAV in smaller aliquots of 20ul for serotype.... The Addgene viral service has now expanded to offer many of our control plasmids in new serotypes, as...optogenetics lie in the spatiotemporal control offered by the system that allows for specific manipulation...H134R) bound to an mCherry reporter. We now also offer AAVrg for plasmid 83898, which targets expression...Gordon Fishell   Chemogenetics Chemogenetics offers noninvasive methods for manipulating cell activity...dependent version of the DREADD hM3D is now also offered as AAV1 (along with AAV PHP.eB, AAV Retrograde,...

  6. Early Career Researcher Toolbox: Social Media for Scientists

    Type
    Blog Post
    ... scientific conference, whether you are attending in person or monitoring that conference via social media...Fast Company, opportunities to speak at conferences, and offers for postdoc positions. Check out the podcast...write a second. While working on this post, I was offered and accepted a position to work remotely as an ...American Society for Cell Biology (ASCB) 2018 Conference. I don’t exactly know how I was selected to participate...s worth a read since many of the pointers are different from the other links shared in the post. And the...

  7. Protein Tagging with CRISPR/Cas9: A Conversation with Mendenhall and Myers

    Type
    Blog Post
    ...and it’s already looking like it’s making a big difference. We can now look at many more transcription factors...Mendenhall: The system is really quite simple but it offers improvements in efficiency and throughput. In adapting... this for tagging any protein in all kinds of different assays, not just transcription factors. The second...quite small—20 amino acids—so we don’t think it interferes with the function of the protein. Myers: We’ve...Mendenhall: The benefits are two-fold: CRISPR/Cas9 offers high efficiency of inducing double strand breaks... surprising to us that there aren’t a hundred different versions of this out there. Addgene: Why was it... They could also switch from FLAG to GFP or a different epitope. Addgene: Do you have any additional tips...

  8. Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins

    Type
    Blog Post
    ...Agrobacterium-mediated DNA transfer, each GOI is usually expressed from a different plasmid, which may lead... entry vectors (pUC57-L1L4 and pUC57-L3L2) that offer the advantage of using restriction digest cloning...plasmids to study PPI using Förster Resonance Energy Transfer (FRET, Heckler et al., 2015) or ratiometric Bimolecular...consists of 4 plasmid toolkits generated with 4 different, binary 2in1 vectors with all possible combinations...change in protein conformation in response to light, offers a way to spatially and temporally control cellular... or dark- state of the photoswitchable proteins offers a powerful approach to control protein-protein ...

  9. Human Germline Editing Using CRISPR

    Type
    Blog Post
    ...al. designed plasmids containing Cas9 and two different gRNAs, and electroporated them with an ssODN repair...experiments in S phase zygotes at 18 hr post fertilization. They observed higher targeting efficiency than...41/42 embryos were non-mosaic at 3 days post fertilization. Since Ma et al. designed a repair template ...about the creation of “designer babies.” The differences in editing between iPS cells and embryos make...off-target effects, but this advantage likely won’t transfer to all genes we’d like to target. In addition,...embryos to only 60%? Unfortunately, given the differences between CRISPR repair in embryos and iPS models...available with current techniques like PGD.  Reference Ma, Hong, et al. "Correction of a pathogenic gene...

  10. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    ...delivery of split base editors Article contributed by Jennifer Tsang AAVs are a great gene therapy delivery mechanism...reproducibility as scientists can assemble parts from different labs and experimental systems in their own research...mutants, determined protein expression levels from different promoters, and inhibited gene expression using...contributed by Erin Sanders CreLite, designed by the Eisenhoffer lab, is a new tool that increases temporal and...bringing the two halves of Cre together. Image from Eisenhoffer et al., 2019. This novel tool is practical...organ culture, as well as cell lineage studies. Eisenhoffer and his colleagues evaluated CreLite for cell...eye, muscle and epithelial cells, as well as in different stages of development, ensuring the breadth and...

  11. CRISPR Meets Synthetic Biology: A Conversation with MIT’s Christopher Voigt

    Type
    Blog Post
    ... to what’s happening at different points within the wood. There are different types of cellulose fibers...number of DNA binding proteins with Cas9 guided by different promoters. The challenge in building genetic circuits...getting enough regulatory proteins that don’t interfere with each other. CRISPR is so orthogonal and programmable...then you’d need the ability to turn on genes at different times. It requires more complex synthetic networks...circuits, they are extremely orthogonal. It’s a different perspective and you can layer them for more complicated...for multi-input CRISPR/Cas9 genetic circuits References: Nielsen AA, Voigt CA. Multi-input CRISPR/Cas...

  12. A Primer on Optogenetics: Introduction and Opsin Delivery

    Type
    Blog Post
    ...numerous variants for the different types of opsins available and their differences and uses have been discussed... the different variants [1, 2, 4]. And, of course, the best way to learn about the many different opsin...about this in detail later). But what are the different steps in an optogenetics experiment and what are...available, instructions for obtaining a material transfer agreement, and list of vendors (such as the vector... core at the University of North Carolina) that offer his constructs. Importantly, control viruses (not... or follow him on Twitter @derekpsimonphd.    References 1. Tye KM, Deisseroth K. Optogenetic investigation...

  13. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    ...mammalian plasmid here and edit on! References and Resources References Kwart, D., Paquet, D., Teo, S. et...cutting efficiency — although both are of course preferred! When trying to strike that balance, you may wonder...sequence constraints, or at least has looser or different requirements. Luckily, there are some available...necessarily more flexible than NGG; they’re often just different (e.g. NGAG, NGAN, etc.). Additionally, alternative...enzymes certainly add more PAM options, none of them offer a one size fits all solution to the issue. Nor are...al). Additionally, SpG Cas9 with the same PAM preference of NG was isolated by the Kleinstiver Lab (read...

  14. Viral Vectors 101: Calcium Sensors

    Type
    Blog Post
    ...remember: Keep Ca+2lm and Ca+2rry on! References and Resources References Nasu Y, Shen Y, Kramer L, Campbell...sensors are commonly used calcium reporters with different kinetic parameters. There are three GCaMP types...and Localization The calcium concentrations in different subcellular compartments can vary greatly. Additionally...measurements are more susceptible to artifacts and pH interference, and comparison between experiments can be more...sites per sensor (to decrease unnecessary calcium buffering and to increase sensitivity), and is a great FRET... also suitable for in vivo studies. There are different versions optimized for neuron-specific, astrocyte-specific...

  15. Antibodies 101: Choosing the Right Isotype

    Type
    Blog Post
    ...antibody for your application!  References and resources References Lu, Q. L., & Partridge, T. A. (1998... while polyclonal antibodies contain a mix of different isotypes and subclasses. They each have their ...of clones and thus the potential to bind many different sites on the target protein they tend to have ...polyclonal primary antibody contains a mix of different isotypes and subclasses, use a broadly reactive... be worth considering an antibody raised in a different species.      Table 1: Isotype considerations...targets, look for primary antibodies raised in different, and distantly related, species, and secondary...

  16. Open Resources and Plasmid Tools For Studying C. elegans

    Type
    Blog Post
    ...collaboration. The field already has comprehensive reference pages and curated databases for scientists including...discussion on the best strategies and protocols for different experiments.  In this blog post let's take a ... peer-reviewed and assigned a DOI that can be referenced. In addition the data published in microPublications...embryonic genes are individually depleted via RNA interference (RNAi). Scientists monitor the effects of RNAi...RNAi using four-dimensional contrast interference (DIC) microscopy and computer imaging processing. The... back frequently for what’s new and popular! References Hammarlund, Marc, et al. "The CeNGEN Project:... biology protocols Read our molecular biology reference ...

  17. How to Prepare for an Industry Interview

    Type
    Blog Post
    ...job hunt, I quickly noticed one of the biggest differences between applying for an academic position versus... phone interview. I also transitioned between different lab groups without having a formal interview process...answer once the interview has started. Instead, I preferred to prepare stories to answer interview questions... an opportunity to demonstrate technical and transferable skills that make you qualified for the position... try and circumvent this, test out the video conferencing software you’ll be using in advance of the interview...interview, if possible. Otherwise, read the video conferencing instructions you received from HR carefully....completing each section of your talk. Some video conferencing software allows participants to “raise” their...

  18. Illuminating Epigenetics with A FRET Based Biosensor

    Type
    Blog Post
    ...in the body contain the same DNA but have very different structure and function, and can have a specialized...syndrome and Prader–Willi syndrome, which have very different disease phenotypes, are two fascinating examples... copies of chromosome 15 are inactivated, but different phenotypes result depending upon whether the improperly...development and progression of cancer through different mechanisms. New FRET based tools to study epigenetics...restriction enzyme digestion, DNA adenine methyltransferase identification, and bisulfite sequencing. ...the reporter consistently showed significantly different FRET levels between unmethylated and methylated... and treatment and improving patients lives. References 1. Ong-Abdullah, Meilina, et al. "Loss of Karma...

  19. Pooled Library Amplifications

    Type
    Blog Post
    ...library protocols you can use as a reference. Table 1: Examples of Different Pooled Libraries at Addgene.  ... plasmids in your library (Are the inserts of different sizes?), and the desired coverage for the library...For this reason, if your library has inserts of different sizes (such as a transcription factor overexpression...your growth time (e.g., 16-18 hours). Finding reference protocols There are many decisions to make when...pDNA Library Amplification protocol are useful references. Skim through our Pooled Library page to find...“one-size-fits-all” protocol! Each library is different, and you should always experimentally test the...

  20. Plasmids 101: Degron Tags

    Type
    Blog Post
    ...you can make an informed choice! References and Resources References Bondeson, D.P., Mullin-Bernstein,... we will review what advantages degron tags can offer over other protein control methods and compare the...tools to manipulate protein expression. These tags offer several advantages over the alternative systems ... all the tags reviewed here, the SMASh tag does offer a few other unique features. If your protein is ... fluorescent Halo ligand can be added to media, buffer, etc., which will bind and light up a tagged protein...profile their effectiveness with over a dozen different proteins. In the end, you can’t predict with absolute...

  21. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    ...proteins that confer affinity for a target – and this serotype can be selected to preferentially target your...voltage indicators reviewed.   References and Resources   References Beck C, Zhang D, Gong Y. Enhanced... These types of sensors can have a variety of different components, but all contain a FRET pair, a genetic... genetically encoded component of the system to offer unique advantages depending on your experimental...powerful tool that swaps capsid proteins from different viruses to optimize them for targeting your cell...

  22. Of Myc and Men

    Type
    Blog Post
    ...explore Myc’s role in cell proliferation and tumorigenesis.  Using two different synthetic peptides corresponding...antibodies, context is key.    References and Resources References Evan GI, Lewis GK, Ramsay G, Bishop... or the C-terminus, Evan, et al. isolated six different monoclonal antibodies (Evan, et al. 1985). Three... found that the N-terminal half of the antigen differs significantly between human and the other species...-9E10 variable regions and sticking them onto different scaffolds. But over the years, several alternative...

  23. Truncated gRNAs for Regulating Gene Expression

    Type
    Blog Post
    ...variants—nuclease-competent or nuclease-null—in order to effect different perturbations. Consider, for instance, the challenge... both cutting and regulation of expression at different genetic loci within a single cell. One might imagine...use of “orthogonal” Cas9 proteins derived from different bacterial species, each of which interacts with... (1). Although this is possible, this strategy suffers from several limitations. Perhaps most importantly...expression (6). Altering the length of gRNAs targeting different sites of interest thus represents a straightforward...introduced together with truncated gRNAs targeting a different set of loci to induce simultaneous cutting and...DNA sequence or regulating gene expression.     References 1. Esvelt, Kevin M., et al. "Orthogonal Cas9 ...

  24. Sharing Your Lab Protocols: Using Apps to Save Time & Track Your Experiments

    Type
    Blog Post
    ...Labguru, Quartzy offers software designed with scientists workflow in mind and can offer a one-stop solution...create lists of your favorite protocols for future reference. The exchange currently boasts over 2,500 protocols... is a subscription-based software, Labguru does offer free trials and pricing options for academic labs... limited to within your lab group, Labguru does offer access to protcols from external vendors, such as...orders. Quartzy caters to life science labs and offers the service free of charge to scientists by allowing...101 articles on various plasmid features, and reference tables (i.e. antibiotic concentrations, commonly...

  25. Great Results Start with Great Standard Curves

    Type
    Blog Post
    ...sample(s) in the same buffer if possible, or very similar buffers if not, since buffer components can affect...non-linear and you should use a different method of analysis. Different fields and assays have varying ...the assay is run, ideally with freshly prepared buffer. This is because standard curves vary depending...range covers a wider span of concentrations but suffers from decreased specificity, especially as it approaches...

  26. Antibodies 101: Producing Recombinant Antibodies

    Type
    Blog Post
    ...antibodies directly into your preferred buffer. If that’s the case, a buffer exchange can be performed using...downstream step, you’ll want to ensure that your buffer components are compatible with your conjugate and...you’ve collected your antibody in the appropriate buffer, it’s time to quantify it. There are several options...our Antibodies 101 blog series.  Resources and references More resources on the Addgene blog Introduction...Purification Protocol Addgene's Antibody Guide References Trimmer JS. Recombinant Antibodies in Basic Neuroscience...

  27. Plasmids 101: Choosing an Antibiotic Resistance Gene

    Type
    Blog Post
    ...decorative" title="Check icon" %}   References and Resources References Hwang, J. M., Piccinini, T. E., Lammel...belongs to the penicillin family of antibiotics. It interferes with cell wall synthesis as its mode of action...beta-lactamase, just like amp is. Thus, the AmpR gene confers resistance to both carb and amp, due to their common...amp), and for some applications, the benefit it offers is marginal. Fortunately, amp can easily be replaced...phosphorylation of kan, which inactivates it. The NPTII gene confers resistance to several aminoglycosides, including...minutes). Pros: Widely available, cost-effective, confers mammalian resistance to G418. Con: Slow transformation...

  28. Don’t FRET: Bimolecular Fluorescence Complementation Makes Visualizing Protein-Protein Interactions Easy

    Type
    Blog Post
    ...Forster Resonance Energy Transfer (FRET). Through the non-radiative transfer for energy between neighboring...requires careful selection of FP fragments and, preferably, some information about the structures of the...level, stability, and subcellular localization may differ from those of your BiFC construct, making it difficult...., do two proteins interact or not? — FRET is preferable when the temporal dynamics of PPIs are the focus...about science whenever he's not off the grid.   References Bischof J, Duffraisse M, Furger E, Ajuria L, ...plasmids and resources Visit our Molecular Biology Reference ...

  29. Plasmids 101: Colony PCR

    Type
    Blog Post
    ...created with the same backbone but that contain different inserts. When you design primers to anneal outside...primer pair to screen for the presence of many different inserts. The downside: this type of primer does... The type of primers you use depends on your preferences. Either way, make sure to test your colony PCR...a standard PCR thermocycling program. One key difference is the plasmid DNA must be released from the ...and without an insert. These controls are quick references you can use when you run your PCR products out... an appropriate molecular weight standard for reference and to add a loading dye with glycerol to your...products have any mutations.  There are lots of different cloning strategies, but regardless of which is...

  30. Cpf1: A New Tool for CRISPR Genome Editing

    Type
    Blog Post
    ...PAM sequence requirements. Cpf1’s preferred PAM is 5’-TTN, differing from that of Cas9 (3’-NGG) in both...endonuclease domain, indicating that Cpf1 functions differently. Since Cpf1 loci are widely distributed across...could be adapted for genome editing. The use of a different nuclease could potentially overcome some of Cas9...pattern opens up the possibility of directional gene transfer, analogous to traditional restriction enzyme cloning... cloning. Sticky-end mediated gene transfer would be particularly helpful for targeting non-dividing cells...shown the diversity of CRISPR-based systems. References 1. Zetsche, Bernd, et al. "Cpf1 is a single RNA-guided...
Showing: 391 - 420 of 1053 results