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We narrowed to 866 results for: FER

Showing: 401 - 440 of 866 results

  1. Will You Be My Mentor? Finding and Asking for Mentoring Support

    Type
    Blog Post
    Published
    Feb. 4, 2014, 3:50 p.m.
    ...to give you advice and guidance. Second, I will offer some advice on how to “make the ask” once you have...have a “posse” of advisors who can help us in different ways and in diverse areas. It is helpful to have...information in these searchable databases are usually offering their contact information so you can tap them ...someone you trust. A good coach can make a big difference, if you can find one. Create your own peer mentoring...with progress. Tell them how their advice made a difference, helped you make a change for the good or resulted...

  2. What's New in CRISPR - November 2021

    Type
    Blog Post
    Published
    Nov. 18, 2021, 2:15 p.m.
    ...mammalian systems has been in question. Three different labs have recently published work that provides... gene activation  and base editing, whereas a different set of mutations (the “V3.1” version) works best...miniature Cas system that utilizes a Cas12f from a different species, Acidibacillus sulfuroxidans, AsCas12f1...They further develop a version of SEND for gene transfer with pseudotyped viral particles, expanding options... the Liu Lab found that mismatch repair (MMR) interfered with the desired resolution of prime editing ...

  3. Plasmids 101: The protein expression toolbox

    Type
    Blog Post
    Published
    Feb. 16, 2023, 7:05 p.m.
    ...protein expression journey! Resources and References References Nabet et al. The dTAG system for immediate...turn off expression for good, the Cre-Lox system offers inducible yet permanent gene excision. Essentially...days. There’s also a CRISPR for that! CRISPR interference (CRISPRi) targets a catalytically dead Cas9 ..., and expression is tunable by titration with different amounts of tetracycline. Tetracycline regulation...

  4. Gendered Innovations: Why Does Sex of the Cell Matter?

    Type
    Blog Post
    Published
    July 21, 2016, 2:30 p.m.
    ...billions—but when drugs fail, they cause human suffering and death. Doing research right has the potential... relevant? Research shows that there are sex differences in the therapeutic capacity of stem cells. Yet...male stem cells. Nor did they detect important differences in function between male and female cells. The...in basic research.  In addition to the studies referenced above, another excellent resource for research...population. To learn a little more about where differences in effects on the sexes come from, one clue or...

  5. CRISPR 101: Cas9 vs. The Other Cas(s)

    Type
    Blog Post
    Published
    Dec. 8, 2022, 2:15 p.m.
    ... put your experiments Nsync.   References and resources   References Dolan, E., Wang, Hou, Zhonggang,...while also highlighting their similarities and differences to Cas9. DNA targeting Cas enzymes Cas9 – The...Originally classified as Cpf1, Cas12’s primary difference from Cas9 is that it generates staggered breaks...(RNA knockdown, RNA editing, etc.). The main differences between Cas7-11 and Cas13 are 1). non-specific...

  6. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    Published
    Jan. 14, 2016, 3:30 p.m.
    ...plasmid) and any other important features that differentiate this plasmid from its predecessor. Important...production and storage, we grow a little extra and transfer 100 µl of glycerol stock to each of 2 96-well ...sequencing provider. Each sequencing service will have different requirements for volume and concentration, and...starts by determining what features you expect to differ between plasmids, and selecting or designing a ...Resources on the Addgene Website View our Plasmid Reference Pages Get Plasmid Backbone Information from Our...

  7. Teaching an Old DOG New Tricks: Controlling Protein Activity with GFP

    Type
    Blog Post
    Published
    Nov. 24, 2015, 3:30 p.m.
    ...GBPb-GAL4 (DNA-binding domain); and a UAS-driven luciferase reporter construct. In the absence of GFP, no...create a complete transcription factor, activating luciferase transcription. Tang et al. coined this system... such that they activate gene expression from different promoters. Moving to an in vivo system, Tang et...eager to see how you use them in your research!  References 1.Tang, Jonathan C.Y., et al. “A nanobody-based...plasmids from this publication at Addgene. 3. Kirchhofer, A., et al. “Modulation of protein properties...

  8. Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them

    Type
    Blog Post
    Published
    May 4, 2021, 1:15 p.m.
    ... we haven’t even imagined yet. References and Resources References Andrews NP, Boeckman JX, Manning ...consist of a mixture of antibodies recognizing many different epitopes. (Middle) Monoclonal antibodies produced...of Recombinant Antibodies Recombinant antibodies offer numerous advantages over polyclonal antibodies and..., Aldred SF, Bestagno M, Burrone O, Maynard J, Ferrara F, Trimmer JS, Görnemann J, Glanville J, Wolf P...

  9. CRISPR Activators: A Comparison Between dCas9-VP64, SAM, SunTag, VPR, and More!

    Type
    Blog Post
    Published
    Oct. 6, 2020, 1:15 p.m.
    ...activation by dCas9-CBP and the SAM system differ in target preference. Sci Rep 9: . https://doi.org/10.1038...programmable guide RNA. Gene activation by dCas9, also referred to as CRISPRa, was initially published in 2013...method, dCas9 is fused to CBP, an histone acetyltransferase domain capable of rearranging chromatin structure...experiment. Good luck with your experiment! References Bikard D, Jiang W, Samai P, Hochschild A, Zhang...

  10. Synthetic promoter AAVs for cell-type specific expression in retinal cells

    Type
    Blog Post
    Published
    March 12, 2020, 1:15 p.m.
    ... Clinical Ophthalmology Basel recently took a different approach to enhance specificity in retinal cells... synthetic promoters, the Roska lab used four different design strategies: Strategy #1 (ProA): Create... this library contained 226 AAVs, each with a different synthetic promoter driving expression of the same...was from a single class of cells  Of the four different promoter designs, promoters from strategy #4 had...ready-to-use AAV viral preps Browse all viral vectors References Juettner, J., Szabó, A., Gross-Scherf, B., Morikawa...

  11. Viral Vectors 101: Chemogenetics

    Type
    Blog Post
    Published
    July 11, 2023, 1:15 p.m.
    ...updated by Jason Nasse in July 2023.  References and Resources References Chen X, Choo H, Huang X-P, Yang X...lead to such responses as decreased heart rate (Redfern et al., 1999) or altered neuron excitability (Zhu...to consider that the same receptor can elicit different responses depending on what cell type it is expressed....sciencemag.org/content/364/6436/eaav5282.long Redfern CH, Coward P, Degtyarev MY, Lee EK, Kwa AT, Hennighausen...

  12. CRISPR 101: Targeting Non-Coding RNAs with CRISPR/Cas9

    Type
    Blog Post
    Published
    Nov. 14, 2024, 2:15 p.m.
    ...difficult to study ncRNA, CRISPR systems offer a number of different options. By carefully considering your...without permanently altering the DNA, CRISPR interference (CRISPRi) and CRISPR activation (CRISPRa) are... you more about its function.  Resources and references Resources on addgene.org Addgene's CRISPR Guide... CRISPR enzymes for transcriptome engineering References Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna...

  13. Quickest Way to Deposit Plasmids: The Deposit Spreadsheet

    Type
    Blog Post
    Published
    March 8, 2017, 3:15 p.m.
    ...easy — our tech transfer team will communicate with your organization’s technology transfer office or other... Send plasmid information to Addgene Material Transfer Agreement (MTA) approval  Send plasmids to Addgene...expression, lentiviral, retroviral, AAV, RNAi, luciferase, cre/lox, yeast expression, worm expression, ...samples to Addgene and contact your technology transfer office about the MTA. Pro Tip! Download a new...

  14. 5 Tips for Troubleshooting Viral Transductions

    Type
    Blog Post
    Published
    Aug. 11, 2016, 2:23 p.m.
    ...concentrating a viral stock. Different titering methods can reveal differences in physical versus functional...will be used in a couple of days, some researchers prefer to store freshly harvested virus at 4 °C instead...up-to-date tips for your virus experiments. References 1. Anthony, Simon J., et al. "A strategy to estimate... Yarmush. "Polybrene increases retrovirus gene transfer efficiency by enhancing receptor-independent virus...

  15. Adenoviral Vector Production and Troubleshooting

    Type
    Blog Post
    Published
    June 14, 2018, 11:44 a.m.
    ... the E1 gene from the packaging cell line is transferred into the AdV by recombination. It is therefore...for production of Gutless AdV can be found in reference 3. Production, amplification, and quality control...please see Addgene’s Adenoviral Guide webpage, and references 1, 2.*Pro-Tip*: once the correct recombinant ...specific for a viral DNA sequence. *Pro-Tip*: Ad5 reference material should be used as an internal control..., high-titer rAdV in your experiment. Enjoy! References 1. Luo, Jinyong, et al. "A protocol for rapid...

  16. Nanoblades: Tiny CRISPR Ninjas for Genome Editing Difficult Cells

    Type
    Blog Post
    Published
    Sept. 26, 2019, 12:50 p.m.
    ... 1-4 different gRNAs. Viral envelope proteins To alter the cell tropism of Nanoblades, different viral...embryos, and animals. Nanoblades can also carry different types of CRISPR cargo including RNPs for indel...can then fuse with the target cell membrane, and transfer their CRISPR cargo into the zygote. As a proof-of-principle...versatile and accessible genome editing tool.   References Mangeot, Philippe E., et al. "Genome editing ...

  17. Mouse Modeling, Part 2: Breeding and Crossing Mice

    Type
    Blog Post
    Published
    Aug. 6, 2019, 12:55 p.m.
    ...separately introduce each mutation independently into different mice, and then these strains should be crossed...When crossed to a mouse that is homozygous for a different mutation, the F1 offspring will be either hemizygous...include embryonic lethality, a short lifespan, or infertility and can arise due to copy number, expression ...transgene. These problems can be mitigated in different ways. Prior to engineering a transgenic strain...engineered mice. Best wishes on your experiment! References Becher, Burkhard, Ari Waisman, and Li-Fan Lu....

  18. Hot Plasmids and Viral Preps - May 2021

    Type
    Blog Post
    Published
    May 18, 2021, 1:15 p.m.
    ...library to identify RBD mutations which do not interfere with ACE2 binding, but impact recognition by specific...immunoprecipitate the entire complex in three different cell lines. The Gräslund lab plans to continue...10.1101/2021.01.30.428974   The CRISPR corner By Jennifer Tsang Here are a few highlights from recent CRISPR...of indels to measure time. These plasmids with different promoters driving expression of Cas9 were used...Janelia Farm GENIE project have arrived! We now offer several versions of GCaMP8 vectors in serotypes ...

  19. Multiplexed Capture of Promoter-enhancer 3D Chromatin Structures Using CRISPR

    Type
    Blog Post
    Published
    July 2, 2020, 12:40 p.m.
    ...compare results across multiple targets because different CAPTURE cell lines will have varying levels of...lines Cell lines, primary cells Table 1: Key differences between original CAPTURE and CAPTURE 2.0 methods...that occur during development and cellular differentiation. Using a cell line model, the Xu Lab tracked...enhancer-promoter interactions during erythroid differentiation. CAPTURE 2.0 captures chromatin's dynamic role....0 re-writes the script on gene regulation. References Han, J., Zhang, Z., & Wang, K. (2018). 3C and...

  20. No Llamas Required - Synthetic Nanobodies Against Membrane Proteins

    Type
    Blog Post
    Published
    June 18, 2020, 1:15 p.m.
    ...randomized with a defined set of different trinucleotides (corresponding to different amino acids) in the regions...this, they injected the spike protein from two different coronaviruses into a llama over several days and...your target protein. The Seeger lab designed 3 different mRNA sybody libraries for use in the protocol,.... Find the Seeger lab's SARS-CoV-2 sybodies! References Geertsma ER (2013) FX Cloning: A Versatile High-Throughput...

  21. Plasmids 101: Protein Expression

    Type
    Blog Post
    Published
    June 7, 2018, 1:17 p.m.
    ...these vectors is controlled by a different promoter resulting in different levels of expression from each...functional, stable proteins. Protein expression refers to the second step of this process: the synthesis...well-defined media and can be easily adapted to fermentation allowing for large-scale, stable production ...labelled or modified amino acids that are useful in different downstream applications. Cell-free expression ...

  22. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    Published
    Sept. 7, 2017, 12:17 p.m.
    ... splitting transfected cells into 3 plates at different densities for selection). A 2% agarose solution...transfection into plates containing selection agent at different densities, then dispose of the plates that do ...freeze at least 2 ampules each of 4 clones with different levels of expression of our gene of interest. ...to rapidly isolate individual cell clones with different expression characteristics from a population of...cell clones picked using the agarose method do not differ grossly from those isolated using cloning rings...

  23. Quick Guide to Working with Drosophila Part 1: Getting Started with Flies

    Type
    Blog Post
    Published
    July 13, 2017, 2:30 p.m.
    ...comes to fruit flies Drosophila start life as fertilized eggs that develop into embryos and then become...about one week from the initial deposition of the fertilized egg by its mother. Inside the pupal case, the... the heat. Isolating male and female flies Differentiating male and female flies is essential for setting...thorax to look like stubble (sb). For a better reference of various fly phenotypes, check out the Classical...latest information about gene function, while others offer fly stocks and other reagents. If you’re ever confused...

  24. Plasmids 101: 5 factors to help you choose the right cloning method

    Type
    Blog Post
    Published
    Aug. 21, 2018, 12:31 p.m.
    ...create customized overhangs to assemble up to 6 different fragments in a prescribed order. These fragments...protocol is fairly flexible and adaptable to cloning different types of DNA, such as cDNA, PCR fragments, or ...process close to 100% efficient. Gateway cloning also offers a very high chance of success (>95% for a single...deposit your finished constructs at Addgene!). References 1. Li, Chaokun, et al. "FastCloning: a highly...Find vector backbones Read our molecular biology reference ...

  25. Plasmids 101: Optimizing Plasmid Yields

    Type
    Blog Post
    Published
    May 26, 2016, 2:30 p.m.
    ... Let it incubate Check the bacterial strain Differences between E. coli host strains can impact plasmid...might want to include an extra wash step or use a different strain. For most applications, Addgene uses and...inadequate plasmid DNA yields. Why would volume make a difference? In the simplest of terms, more volume = more...conditions play a critical role in plasmid DNA yield. Different plasmids and strains will vary in their optimal...media with supplements such as magnesium salts, buffering agents, and/or supplying additional carbon sources...

  26. When is a Monomer not a Monomer? The Top Three Ways Your Favorite Fluorescent Protein Oligomerizes in Cells

    Type
    Blog Post
    Published
    April 19, 2016, 2:30 p.m.
    ... alter the ability of one FP to nonradiatively transfer energy to the second FP, often decreasing the ...most cases, by using monomeric FPs or FPs from different families, which cannot form heterodimers (i.e....University, did his postdoctoral fellowship with Jennifer Lippincott-Schwartz at the National Institutes...avid long distance runner, gardner, and cook.   References 1. Snapp, E.L., et al., Formation of stacked ...Protein Resources Find Other Plasmids that Glow (Luciferase Plasmids) Find Optogenetics Resources ...

  27. Anatomy of a Plasmid Page at Addgene

    Type
    Blog Post
    Published
    Feb. 4, 2016, 3:30 p.m.
    ... ever found yourself bamboozled by all of the different kinds of information on our plasmid pages? Well...depositing lab, although you may have an entirely different use in mind! This section of our plasmid pages... you to use in your Materials and Methods and References sections when citing the use of your colleague...backbones.  The backbone modifications listed can refer to changes made to the original backbone by the ...e.g. if it was derived from a plasmid made in a different lab - or one obtained from Addgene!).  It also...

  28. Plasmids 101: Multicistronic Vectors

    Type
    Blog Post
    Published
    Sept. 9, 2014, 8:20 p.m.
    ...600 bp) and may take up precious space in viral transfer vectors with limited packaging capacity. Additionally...placement of the reporter gene). Addgene's collection offers a variety of plasmids to express two or more genes...Retroviral pEF1a-IRES-Neo IRES Mammalian MSCV-IRES-Luciferase IRES Retroviral pWPI IRES Lentiviral AmCyan-P2A-mCherry... new eBook! Our goal was to create a one-stop reference guide for plasmids. We've combined our Plasmids...solutions necessary for advancing the field. References: 1. Internal initiation of translation of eukaryotic...

  29. Optogenetics + CRISPR, Using Light to Control Genome Editing

    Type
    Blog Post
    Published
    Sept. 3, 2020, 12:15 p.m.
    ...CRISPR gene activation system, but settled on a different optimal fusion protein combination. The optimized...AcrIIA4, this unfolding conformational shift can interfere with activity. The resulting system was coined...Alexander Deiters’ lab, on the other hand, took a different approach. These researchers used a genetically...expression - demonstrating light-induced silencing of transferrin receptor CD71 in HeLa cells.   Comparison of...applications you use these for in the future! References Bubeck F, Hoffmann MD, Harteveld Z, Aschenbrenner...photoactivatable transcription systems to induce neuronal differentiation. Nat Methods 14:963–966 . https://doi.org/10.1038...

  30. A Guide to Starting Your Own Journal Club

    Type
    Blog Post
    Published
    Feb. 6, 2020, 3 p.m.
    ...the same figure by figure organization with a different person responsible for each figure. Others still...questions they have.  These variations are suited for different applications and should be carefully chosen for... and audio recordings of the meeting using teleconferencing software. Any file sharing service should ...or instant messaging but we as scientists often prefer data. At Addgene we rely on polls and surveys. ...acquisition of the essential data and allow everyone to offer thoughts you may not have even considered. These...

  31. An Introduction to Adenovirus

    Type
    Blog Post
    Published
    Oct. 3, 2023, 1:15 p.m.
    ...written by Addgenie Brian O'Neill.  References and resources References Bauzon, M., Castro, D., Karr, M.,...vectorization strategies include creating an AdV transfer plasmid with the E1 and E3 (Early 1 and Early ...Although the late genes are usually present in the transfer plasmid for the majority of adenoviral vector ...B. (2014). Adenoviruses: General Features. In Reference Module in Biomedical Sciences. Elsevier. https...

  32. Course-Based Undergraduate Research Experiences (CUREs)

    Type
    Blog Post
    Published
    Feb. 9, 2023, 2:15 p.m.
    ... in and out of the classroom.   References and Resources References Bangera, G., Brownell, S. 2014. Course-based...the University of San Diego), decided to take a different approach. His own most impactful learning experiences...all shaped his teaching style, with the former offering excellent training in improvisation and the latter... available technical and pedagogical resources, offers mentoring for instructors, and provides opportunities...

  33. CRISPR Between the Genes: How to Experiment with Enhancers and Epigenomics

    Type
    Blog Post
    Published
    Aug. 9, 2016, 2:30 p.m.
    ...denotes intersection and the – operator denotes set difference. That’s the intuition for how genome math helps...enhancers from step 2. See which ESCs die or differentiate Step 3 reveals which NANOG-related genes are...distinct enhancer disrupted. As our ESCs die and differentiate over time, we periodically use next-generation...Therefore if a gRNA drops or disappears over time, we infer that the enhancer it targets is a “pillar of function...by starting with all of the DHS sites from 125 different human cell types from the ENCODE Project. DHS ...

  34. Simplify Cloning with in vivo Assembly

    Type
    Blog Post
    Published
    Oct. 18, 2018, 12:37 p.m.
    ...This intrinsic bacterial recombination pathway, referred to as recA-independent recombination, joins together...single PCR, for example for subcloning genes from different templates. All PCRs are run as an 18 cycle, 25...pre-made aliquots of PCR master-mix, containing buffer, polymerase and dNTPs, can be stored frozen: just... on Twitter @jakefwatson and @JGarciaNafria.  References 1. Jones, Douglas H., and Bruce H. Howard. "A...for plasmid cloning Find our molecular biology reference here Find molecular biology protocols here   ...

  35. INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons

    Type
    Blog Post
    Published
    Dec. 7, 2020, 2:53 p.m.
    ...twitter @Vopinator.     References: Halpin-Healy TS, Klompe SE, Sternberg SH, Fernández IS (2020) Structural...Insertion of the mini-transposon can occur in two different orientations, but is strongly biased towards T-RL...combined in single-plasmid INTEGRATE (pSPIN), which offer simple transformations for smaller cargos, and the...integration efficiencies achieved with VchINT, at two different incubation temperatures. Data adapted from Vo ...

  36. Minigenomes - a Safe Way to Study Dangerous Viruses Like the Ebola Virus

    Type
    Blog Post
    Published
    March 17, 2016, 2:30 p.m.
    ...viral infection), producing firefly luciferase. Firefly luciferase can readily be measured to quantify...single (non-viral) reporter gene (e.g. firefly luciferase or eGFP) flanked by the EBOV-specific gene start...to transcribe the reporter gene (e.g. firefly luciferase). The polymerase recognizes the gene start signal...transcription of negative strand RNA viruses.   References 1. Buchholz, Ursula J., Stefan Finke, and Karl-Klaus...

  37. Hot Plasmids - August 2020

    Type
    Blog Post
    Published
    Aug. 18, 2020, 1:15 p.m.
    ...human pluripotent stem cells By Jennifer Tsang CRISPR gene interference (CRISPRi) and gene activation (...For labeling the plasma membrane, they tested different target sequences for improved selectivity and ...into the supernatant. Four plasmids containing different nanobodies (NB) were generated in pMAK:  pMAK461....1038/s41598-020-57500-1 The CRISPR corner By Jennifer Tsang Here are a few highlights from recent CRISPR...

  38. Tips for CRISPR Gene Editing in Mice

    Type
    Blog Post
    Published
    June 28, 2016, 10:59 a.m.
    ... to determine the pros and cons of using many different types of gRNA design software. When you are happy...inserted or deleted. Each lab will use a slightly different method to genotype their pups. Taking a toe, a...weeks old and use the ear tag method (creating different patterns of ear holes and using that material ...latest and greatest in scientific content.      References 1. Cho, Seung Woo, et al. "Analysis of off-target...plasmid into zygotes." Development, growth & differentiation 56.1 (2014): 122-129. PubMed PMID: 24372541...

  39. CRISPR 101: Targeting RNA with Cas13a (C2c2)

    Type
    Blog Post
    Published
    Sept. 21, 2017, 2:07 p.m.
    ...limitations associated with SpCas9. Cas13a (previously referred to as C2c2), has several unique properties that...for target recognition and cleavage). C2c2 (now referred to as Cas13a) is unique in terms of structure ...remainder of this blog post. Perhaps the biggest difference between Cas13a and Cas9 is that Cas13a binds ...SHERLOCK (depicted in figure 1) has been used to differentiate strains of Zika virus, genotype human DNA, identify...Gearing contributed to the update of this post. References 1. Abudayyeh, Omar O., et al. "C2c2 is a single-component...

  40. Deciphering the Mysteries of Behavior with Viral Vectors

    Type
    Blog Post
    Published
    Sept. 17, 2019, 12:40 p.m.
    ...moving around to explore the environment has a different motivation (and brain areas associated) compared...addiction. Anxiety tests Rodents have a natural preference to stay in dark areas (protected) as opposed ...to brightly illuminated ones (risky). They also prefer to explore novel environments by initially moving...anxiety. On the other hand, the control group preferred staying in the dark compartment (Howerton at al...these in a  future blog post, so stay tuned!  References Alpár, Alán, et al. "Hypothalamic CNTF volume...
Showing: 401 - 440 of 866 results