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We narrowed to 866 results for: FER

Showing: 361 - 400 of 866 results

  1. Cre-ating New Methods for Site-specific Recombination in Drosophila

    Type
    Blog Post
    Published
    May 12, 2015, 1:32 p.m.
    ...the efficiency of Cre recombination varies for different constructs and cell types. Second, Cre may induce...from the standard UAS/GAL4 system is toxic to proliferating cells. A Cre-estrogen receptor ligand binding...enhance genome editing capabilities in Drosophila. References: Multiple new site-specific recombinases for ...for use in manipulating animal genomes. Nern A, Pfeiffer BD, Svoboda K, Rubin GM. Proc Natl Acad Sci USA...PubMed. Reduction of Cre recombinase toxicity in proliferating Drosophila cells by estrogen-dependent activity...

  2. Celebrating Outstanding CRISPR/Cas9 Achievements at the Dr. Paul Janssen Award Dinner

    Type
    Blog Post
    Published
    Sept. 15, 2014, 6:05 p.m.
    ...Dr. Jennifer Doudna and Dr. Emmanuelle Charpentier. I was joined by Addgene Scientist Matt Ferenc (one...especially excited to attend this event because Jennifer did her graduate work one floor above mine in ...system Drs. Dounda and Charpentier met at a 2011 conference and decided to collaborate on what was a “curiosity... science to the non-scientists in the room. He offered the great analogy of comparing the CRISPR/Cas9 ...

  3. Lentiviral CRISPR Libraries Enable Genome-Scale, Knockout Screening

    Type
    Blog Post
    Published
    Feb. 20, 2014, 4:57 p.m.
    ...online on Addgene's CRISPR Library Reference page.    References: Genetic Screens in Human Cells Using...GeCKO library, each library has been enriched for different gene targets. These pooled libraries include enrichment...lentiviral libraries and their demonstrated application offer proof-of-principle that CRISPR-Cas9 can be used ...any sequence in the genome. Tips and tricks Wang offers this tip: “It's quite important, at every step ...

  4. Tips for a 1st Time CRISPR User (by a 1st Time CRISPR User)

    Type
    Blog Post
    Published
    March 7, 2017, 3:30 p.m.
    ...at how variable Cas9-expression was across the different cells (Figure 3).   Figure 2: Cas9 expression...publication. Pro-Tip: 1. Use multiple gRNAs to target different regions of your gene. I used existing BRAF gRNA...our website. There are a few reasons to target different regions of the gene with multiple gRNAs, but the...comparing the phenotypes of multiple gRNAs. Targeting different regions of the gene is also useful in case the...that would co-migrate on a gel) instead of two different sized digestion products (which could be easier...your way to making new scientific observations. References 1. Rago C, Vogelstein B, Bunz F. Genetic knockouts...

  5. Some Like it Hot: Thermostable GeoCas9

    Type
    Blog Post
    Published
    Sept. 14, 2017, 12:40 p.m.
    ... just find one that naturally occurs? This is different than the engineered approach used to make eSpCas9...SpCas9 or GeoCas9 RNPs were incubated in reaction buffer at temperatures ranging from 25-80 °C. Activity...incubated in human plasma diluted with reaction buffer for 8 hours at either 37 °C or 4 °C. Substrate ...gel. SpCas9 is stable when incubated in reaction buffer alone, but has limited activity when incubated ...are available from Addgene. Happy CRISPRing! References Harrington, Lucas, et al. "A thermostable Cas9...

  6. Plasmids 101: What is a plasmid?

    Type
    Blog Post
    Published
    April 2, 2020, 6:17 p.m.
    ...fundamentals. Our mission is to curate a one-stop reference guide for plasmids, so that you can spend less...their artificial nature, lab plasmids are commonly referred to as “vectors” or “constructs.” To insert a gene...Green Fluorescent Protein (GFP) Plasmids 101: Luciferase Monitor the level of a chemical in a particular...plasmid use in the lab. Addgene’s Molecular Biology Reference includes information about molecular cloning, ...a plasmid vector, molecular biology tools and references, and how to maintain your plasmid stocks. The...

  7. Plasmids 101: Gene, enhancer, and promoter trapping

    Type
    Blog Post
    Published
    May 2, 2023, 1 p.m.
    ...tools for your next experiment! References and Resources References Wood, V., et al., Hidden in plain...an endogenous gene. Each trap uses a slightly different approach (described in detail below), but fundamentally...kidneys at a specific developmental stage you could infer that the gene which GFP was monitoring was being... these circumstances, trap methods may be the preferred screening option. Promoter traps Promoter trap...

  8. Cellosaurus: A Cell Line Information Database

    Type
    Blog Post
    Published
    Feb. 21, 2023, 2:15 p.m.
    ...specific compounds, links to publication references, and cross-references with cell line catalogs and collections...be adjusted based on query, reference, and shared allele search preferences to help you identify proximal...to learn more, check out the Cellosaurus FAQ! References Lorsch JR, Collins FS, Lippincott-Schwartz J....

  9. Antibodies 101: Affinity Tags

    Type
    Blog Post
    Published
    Sept. 19, 2023, 1 p.m.
    ...antibodies, and more!   Resources and References   References Coligan, J. E., Dunn, B. M., Speicher, ...blot and immunofluorescence. GST  Glutathione S-Transferase (GST) is a 26 kDa tag which can be isolated with...Nonetheless, this tag can be purified with several different resin types and is a fairly small tag that is ...  Affinity tags vs. other tags There are many different types of tags, including epitope tags (FLAG, Myc...

  10. Running for Rare Disease, Running for FOP, Running for AJ

    Type
    Blog Post
    Published
    Sept. 15, 2016, 2:30 p.m.
    ... with a six-year-old boy named AJ Gonzales who suffers from Fibrodysplasia ossificans progressiva (AKA...first met AJ and his family, he did not seem much different than my second son of similar age. When he came...therapeutic antibody that binds Activin A and interferes with inappropriate activin-ACVR1 interactions...superpower for those running on behalf of rare disease sufferers like AJ. As the Blue Lantern Oath says: "In fearful...running on behalf of AJ Gonzales, a young boy suffering from FOP, raising money for NORD for the past ...

  11. What Do I Do Now? Academic v. Non-Academic Career Decisions

    Type
    Blog Post
    Published
    March 9, 2017, 3:40 p.m.
    ...encompasses millions of choices including pharma, tech transfer, management consulting, science communication,...of today’s postdoctoral training can directly interfere with a seamless jump into industry.” In his article...interact with scientists just like you What is different about academia? Publishing in peer reviewed journals...Constant worry about grants and renewals What is different about non-academia? Scientific publications are...where you want to go will help you focus on the transferable skills that you will need to get there. Additional...

  12. CRISPR Challenges: Standardization and Homology Directed Repair

    Type
    Blog Post
    Published
    Feb. 14, 2018, 2:36 p.m.
    ...work demonstrates that different repair template strategies activate different HDR sub-pathways like synthesis-dependent...live updates! My most popular tweet from the conference is shown above. The blog post I mentioned, John...post-CRISPR cleavage When I first looked at the conference agenda, I was surprised to see that it would ...Having had time to digest my notes, since the conference, I think I may have finally grasped just how ...

  13. Identifying Sequence Elements with SnapGene's Feature Database

    Type
    Blog Post
    Published
    Dec. 21, 2017, 2:06 p.m.
    ...due to genetic drift and the use of genes from different microbial strains. It proved to be impractical... least 96% sequence identity when detecting a reference feature. For a coding sequence feature that may... could miss a common feature if the sequence differences exceed the threshold. This issue is addressed...often come in closely related versions that have different properties. To prevent misidentification, we augment...way for SnapGene users to apply features from a reference strain to newly sequenced strains through our ...

  14. HA Frankenbody, a New Imaging Tool to Visualize Single Molecules and Nascent Peptides

    Type
    Blog Post
    Published
    Oct. 10, 2019, 12:41 p.m.
    ...antibodies for tagging purposes. Different antibodies bind different protein sequences called epitopes...infamous frankenstein, was constructed by grafting different parts together. By combining two optimized elements...living cells. To make these scaffolds specific to different targets, the scientists fused them to sequences...to build a more robust set of frankenbodies. References Ewert, S., Honegger, A., & Pluckthun, A. (2004...

  15. New Viral Vectors - Winter 2025

    Type
    Blog Post
    Published
    Jan. 28, 2025, noon
    ...Request service, which offers over 3,000 eligible transfer plasmids and five different serotypes. Not sure...pAAV_hSyn1-SIO-stGtACR2-FusionRed AAV5 Optogenetics Ofer Yizhar New serotype pAAV-CaMKIIa-ChRmine-oScarlet-KV...pAAV-CaMKIIa(0.4)-eOPN3-mScarlet-WPRE AAVrg Optogenetics Ofer Yizhar New serotype pAAV-syn-Flpo AAVrg Recombinase...

  16. Viral Vectors 101: Transductions

    Type
    Blog Post
    Published
    April 30, 2024, 1:15 p.m.
    ...transductions. If you’ve ever wondered how they are different from or similar to the other Ts, we have your ...transduction?  Transduction is the process of transferring foreign nucleic acid into a cell by a viral ...controlled lab usage. Using viral transductions can offer benefits such as tissue-specificity, high efficiency...that you can swap out your virus type to achieve different biological outcomes. For example, some viruses...target cells or organism, some primary cells and differentiated cell types are picky when it comes to transfection...

  17. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    Published
    Sept. 21, 2021, 1:15 p.m.
    ...cell-free transcription/translation systems (also referred to as TX-TL systems) have been gaining popularity...initial system by using a stronger promoter and different biochemical secondary energy sources to increase...XMAS-TREE. a) Cells are transfected with three different vectors and flow cytometry is used to sort populations...XMAS-TREE-based methods at various genomic loci in four different categories.     Brookhouser, N., et al. BMC...10.1186/s12915-020-00929-7    The CRISPR corner By Jennifer Tsang New CRISPR plasmids are always being added...

  18. A New Optogenetic Tool Based on AraC Controls Gene Expression with Blue Light

    Type
    Blog Post
    Published
    June 17, 2021, 1:17 p.m.
    ...Khammash lab can also be found here.   References and Resources References Romano E, Baumschlager A, Akmeriç...functional BLADE variants, too. The team tested different linkers between VVD and the DNA-binding domain...the Molecular Biology Reference Download Addgene's Plasmids 101 eBook ... Jennifer Tsang...

  19. Extracurricular Activities for a Strong Science Career Path

    Type
    Blog Post
    Published
    Jan. 21, 2014, 3:08 p.m.
    ...However, careers outside of the lab highlight “transferable” skills such as leadership, management, and ... and trends “Never Leave an Empty Seat” at conferences and seminars, as this is a valuable time to network...particular field of study Assess your interest in a different field Outreach activities (e.g. Volunteer...Leadership Networking University technology transfer office positions   Market analyses Patent...Marketing Communication Business models Conference or club organizer Management Leadership...

  20. Hassle-free 96-well Format Epitope Tagging Using Cas9 Ribonucleoprotein

    Type
    Blog Post
    Published
    June 28, 2018, 3:01 p.m.
    ...numerous cell lines can be generated, each with a different tagged protein of interest, and a universal all-purpose.... Current methods of epitope tagging, however, suffer from many pitfalls: they require construction of... achieve 5-30% tag knock-in efficiency across different stem cell types. This has facilitated easy generation...format and tested parallel tag knock-in for 96 different genes in a single go (Figure 3). We were able ...optimization includes selection of transfection buffer and electroporation pulse program to achieve high...seen daydreaming about sunny days in the UK.   References 1. Bressan RB, Dewari PS, Kalantzaki M, Gangoso...

  21. Quick Guide to Working with Drosophila Part 2: Controlling Gene Expression in Flies with Gal4/UAS

    Type
    Blog Post
    Published
    July 21, 2017, 12:48 p.m.
    ... Gal4 lines and study gene expression in many different tissue types! Although putting a gene of interest... overexpressed in several cell types by using different Gal4 drivers. It is much easier to mate flies ...one gene in one cell type and another gene in a different cell type (6–8). Although it’s often very easy...the meantime, be sure to browse the publication references and web links from my first post and start designing...jonchowphd but he still reads “#” as a “pound symbol”. References 1. A. H. Brand, N. Perrimon, Targeted gene expression...

  22. New and Upcoming Viral Vectors - June 2019

    Type
    Blog Post
    Published
    June 18, 2019, 1:15 p.m.
    ...patterns of different AAV serotypes. To help make this easier for scientists, we now offer trial size ...excited to offer these useful tools to the community. Among other new biosensor tools, we now offer the jGCaMP7c...control of specific neuronal populations. We now offer a Flp-expressing AAV, as well as a Cre-dependent...

  23. Unique and persistent IDs for improved reproducibility: Addgene now supports RRIDs and compact identifiers for all plasmids

    Type
    Blog Post
    Published
    Nov. 20, 2018, 8:17 p.m.
    ...identifiers (PIDs), a long-lasting way to identify and reference documents, files, or physical reagents. What ...and exist online indefinitely. PIDs historically referred to objects that exist online, but this approach...you, we’ll go through examples to highlight the difference between them using a plasmid available through...the lenti-Cas9-VQR plasmid page. Despite their differences, both of these PID formats for lenti-Cas9-VQR...managed by Addgene. How to cite options using the different PID formats The instructions for how to cite any...

  24. Networking Follow-up: I Met a Person. Now What?

    Type
    Blog Post
    Published
    Jan. 12, 2021, 2:15 p.m.
    ...events for one organization or go to the same conferences/meetings over and over to get to know the regulars...people by discussing non-work topics and interests. Offer help when you can, instead of just asking for favors...starfish shape. You can ask me about the different arms of this starfish sometime, but as of yesterday...come across your contacts over the years in many different ways. This leads me to my last tip: It’s usually...Browse other blog posts on networking Attending a conference soon? Get tips here Learn about informational...

  25. DAP arrays for multiplex gene editing

    Type
    Blog Post
    Published
    July 7, 2022, 1:15 p.m.
    ...heart disease, and type 2 diabetes, across four different types of human cells, including T cells and hematopoietic... interest with only minimal genetic payloads. Different from existing multiplex array architectures, including...when repetitive tRNA sequences must be avoided, different tRNAs with optimized 5’ leader sequences can be... lab of Xue Sherry Gao at Rice University.    References and Resources More resources on the Addgene blog...Adding Precision and Flexibility to CRISPR Editing References Yuan Q, Gao X (2022) Multiplex base- and prime-editing...

  26. Using CRISPR/Cas9 to Edit Disease Out of the Genome

    Type
    Blog Post
    Published
    June 23, 2022, 1:15 p.m.
    ...seeing adoption of the technology in different labs for different applications.” Li’s team noted that ...cells of patients with cystic fibrosis (8).   The difference with CRISPR/Cas9 in comparison to other genome...” said the University of California Berkeley’s Jennifer Doudna. “They are the first direct demonstration...possible through the use of nanoparticles. [6].    References 1. Zhang B. 2021. “CRISPR/Cas gene therapy.” ...

  27. Revamp Your Lab Meetings With Creative Virtual Collaboration

    Type
    Blog Post
    Published
    Feb. 9, 2021, 2:15 p.m.
    ...for creative interaction During Zoom meetings, different tools can be used to promote creative interaction...as a picture. I believe that showing something different from time to time breaks up the monologue a little...papers will be written by several authors from different institutes, disciplines, and cultural backgrounds...and only general chatter will be allowed with a different host each time that moderates conversations. This...communities. You can follow him on twitter @salmon_phd References Frassl MA, Hamilton DP, Denfeld BA, de Eyto E...

  28. Hot Plasmids - December 2019 - CRISPR pooled library, LipoGlo, CRISPRa, and Voltage Imaging Tools

    Type
    Blog Post
    Published
    Dec. 10, 2019, 1:58 p.m.
    ... that affect Th2 differentiation and activation, using expression of differentiation factors as the screen...ApoB-NanoLuc reporter by inserting the NanoLuc luciferase downstream of the apoB gene. Using this reporter... imaging can be performed in intact animals and offers many advantages over traditional measurements such...Gottschalk group hypothesizes that this may be due to different lipid compositions of C. elegans membranes, which...

  29. Plasmids 101: NGS Quality Control for Pooled Libraries

    Type
    Blog Post
    Published
    Oct. 26, 2017, 1:59 p.m.
    ... the library are differently sized, smaller plasmids may be amplified preferentially, skewing the composition...some of Addgene’s most exciting and versatile offerings! We recently re-amplified our distribution stock...successful amplification? To provide a point of reference, Addgene commonly prepares NGS samples using both...overrepresented plasmids (plasmids that were amplified preferentially). You can also rank the plasmids by abundance...

  30. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    Published
    Feb. 27, 2024, 2:15 p.m.
    ... will be depositing multiple plasmids and would prefer to send us your plasmid information in spreadsheet...plasmids all built with the same backbone and only differing in the gene/insert. Pooled libraries are normally...click the "Save" button and jump around to the different pages by clicking on the appropriate boxes at ...at the bottom of the page before moving onto a different page so that any data you enter is not lost.   ...to the backbone, and the vector type. Pro Tip! Reference the Addgene ID in the "Backbone Manufacturer" ...concentration of 0.1–1 µg/µL (or bacterial streaks, if you prefer). If you wish to edit your plasmid data after submission...

  31. Cancer, Inflammation and Immunity - Harnessing the Body’s Defenses to Fight Cancer

    Type
    Blog Post
    Published
    July 6, 2017, 1:59 p.m.
    ...organizers. My overall impression from this conference was that Cancer Researchers and Immunologists...Here I aim to compile a few highlights from the conference which reflect how cancer/immunology researchers...multi-pronged attack. Inflammation and tumorigenesis The conference opened with a very interesting presentation by...our Fluorescent Protein Guide. Throughout the conference aspirin and the use of COX inhibitors was discussed...about our free deposit and distribution service! References: 1. Pietrocola, Federico, et al. "Caloric restriction...

  32. Screening for Successful Genome Editing with Digital PCR

    Type
    Blog Post
    Published
    March 30, 2017, 2:30 p.m.
    ...mutation of your favourite gene, there are several different methods available. Thankfully, there are many ...mutations. The assays you will use aren’t that much different from conventional duplexed primer probe assays...primer to amplify the target locus, as well as a reference probe designed to bind at an unedited site distal...are completely “blind” to this very important difference (Figure 3). If you are interested in harnessing...biology. Follow him on Twitter @AnotherLabRat.     References 1. Miyaoka, Y. et al. Isolation of single-base...

  33. Top Requested Lentivirus and AAV of 2016

    Type
    Blog Post
    Published
    Jan. 6, 2017, 3:56 p.m.
    ...AAV8) enable different patterns of transduction and transgene expression across different cell types (...world. We began with only a few inventory items offered domestically, but by the end of 2016, we expanded...visualizing hM4D(Gi)-mCherry in fixed tissue. References 1. Aschauer DF, Kreuz S, Rumpel S. Analysis of...Kilic E, Bähr M. Human synapsin 1 gene promoter confers highly neuron-specific long-term transgene expression...

  34. 15 Years of Addgene: The Top 15 Plasmids

    Type
    Blog Post
    Published
    Jan. 8, 2019, 1:54 p.m.
    ...illustrates the utility these tools have in many different areas of biology as they grow. The top 15 plasmids...envelope protein enables this virus to infect many different cell types so it is useful in a wide range of ...lentiCRISPR v2 - This 3rd generation lentiviral transfer vector also from the Zhang lab is used to deliver... David Root lab is a 3rd generation lentiviral transfer plasmid for cloning and expressing new shRNA sequences... Jennifer Tsang...

  35. Flow Cytometry Readouts: Yes, No, and Everything in Between

    Type
    Blog Post
    Published
    Aug. 20, 2024, 1:15 p.m.
    ...Pro tip! If your samples and control underwent different treatment processes, you may see a shift in the...presence or absence, the plots can look quite different. You might have high expression, low expression... the mean fluorescence intensity (MFI) of the different populations. MFI is the average brightness from...plot. However, the histogram plots look a bit different than they do for yes-no readouts.      ...above is for a smeary expression and is quite different from the yes-no histogram plot shown previously...

  36. Plasmids 101: Screening Strategies Used in Plasmid Cloning

    Type
    Blog Post
    Published
    Sept. 15, 2020, 1:15 p.m.
    ...You’ll want to pick enzymes that will give you different sized bands depending on whether the plasmid contains...using vector-specific primers, look for size differences. You can also use primers where one primer anneals...popular sequencing primers in our Molecular Biology Reference. Sometimes common  primers can't provide all of...Sanger sequencing Read the Molecular Biology Reference ... Jennifer Tsang...

  37. Fluorescent CRISPR Reporters: SRIRACCHA and GEmCherry2

    Type
    Blog Post
    Published
    May 26, 2020, 1:15 p.m.
    ...These cells are then transfected with several different constructs for the reporter to work and to make... visual read out allows for easy screening of different gRNAs by simply changing the gRNA used when transfecting...concept the Denham lab compared editing at three different genomic sites in the AAVS1 locus using GEmCherry2...GEmCherry2. In this experiment they saw a clear difference in mCherry expression using FACs sorting, confirming...mutant of interest, give these methods a try.  References Højland Knudsen C, Ásgrímsdóttir ES, Rahimi...

  38. Building and Validating Recombinant Antibodies: A Case Study

    Type
    Blog Post
    Published
    April 5, 2022, 1:20 p.m.
    ...commonly used in research laboratories to detect the different subclasses, but there are also secondary antibodies...secondary antibodies to detect multiple targets with different mAbs in the same experiment. The Trimmer Lab found...within the plasmid backbone to generate R-mAbs of different subclasses, including subclasses that were not... useful research materials and information. References and Resources Additional Resources on the Addgene...Addgene's recombinant antibody plasmid collection References Andrews NP, Boeckman JX, Manning CF, Nguyen JT...

  39. Sweating the Small Stuff: Details in the Lab

    Type
    Blog Post
    Published
    Feb. 8, 2022, 2:15 p.m.
    ...the answer isn't to work harder - it's to work differently.  Tools to Try  Checklists Found in surgical ...and your workflow can have a huge impact. Test different approaches, like whether it's better for you to... more than anything else I tried, made a huge difference in my ability to focus, especially on days when...what can be a rather vicious cycle.  Everybody’s Different   The truth is, there is no One True Way to Error-Free...the detail-driven daily grind of science.      References and Resources Images used Roberson Lab, UPenn...

  40. Plasmids 101: Methylation and Restriction Enzymes

    Type
    Blog Post
    Published
    June 30, 2016, 2:30 p.m.
    ...DNA. The other component of these systems, methyltransferases, methylate DNA at particular sequences to...restriction modification systems containing methyltransferases that add methyl groups to specific DNA sequences...contain 3 methylases that recognize and methylate different stretches of DNA: Dam methylase adds a methyl...plasmids with no methylation (there are no methyltransferases in the PCR reaction). The template plasmid...could overlap with a methylation site. The quick-reference table below lists 10 common enzymes that may be...
Showing: 361 - 400 of 866 results