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  1. Important Considerations When Using AAVs

    Type
    Blog Post
    Published
    June 13, 2017, 2:30 p.m.
    ...still be taken when using them. There are suspected cases of insertional mutagenesis in humans (1) and AAVs...Currently, AAVs are being used to deliver CRISPR/Cas9 but have the potential to deliver other therapeutic...used with new exciting technologies (such as CRISPR/Cas9) as they allow for gene delivery in an efficient... 24086725. PubMed Central PMCID: PMC3785459. 4. Castle, Michael J., et al. "Adeno-associated virus serotypes...Expression in Mammals A Match Made in Heaven: CRISPR/Cas9 and AAV Viral Vector Elements Additional Resources...

  2. Synthetic Biology, Artificial Chromosomes, iGEM & More - Interview with Tom Ellis

    Type
    Blog Post
    Published
    Nov. 29, 2017, 2 p.m.
    ...Today’s episode of the Addgene Podcast features guest host and Addgene European Outreach Scientist, Benoit...researcher. Listen to the interview with Tom Ellis Podcast breakdown Intro: How Tom ended up in academia and...the audio engineer and producer for the Addgene Podcast.      Additional Resources from the Addgene Blog... addgene podcast...

  3. Custom CRISPR Screens & the Green Listed Software

    Type
    Blog Post
    Published
    July 11, 2017, 2:30 p.m.
    ...used for each gene. At the moment, 13 full genome SpCas9 knockout libraries designed by different academic...Library, for example related to strategies beyond SpCas9, or species not currently included in Green Listed...libraries. As of now I have focused on human and mouse SpCas9 knockout libraries, but there are of course great...activity and minimize off-target effects of CRISPR-Cas9." Nature biotechnology 34.2 (2016): 184-191. PubMed...PMC5445660. 7. Chari, Raj, et al. "Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library...

  4. Addgene-Seeding Labs Plasmid Grant: Accelerating Science Globally - 5 Scientists, 4 Countries & 3 Continents!

    Type
    Blog Post
    Published
    July 18, 2017, 12:41 p.m.
    ...Optogenetic Tools Like pDawn  Making cassava plants resistant to cassava mosaic virus Dr. Fiona Robertson,...and Biotechnology expert. In order to preserve Cassava yields and to increase the potential for this drought...plant, Dr. Robertson aims to make use of the CRISPR/Cas9 technology to engineer plants that are able to destroy...

  5. Hot Plasmids and Viral Preps - September 2021

    Type
    Blog Post
    Published
    Sept. 21, 2021, 1:15 p.m.
    ...including several pooled libraries: Cas13X and Cas13Y, two compact Cas13 proteins found in the metagenome...end-point sequencing assays. These plasmids use “Cas9-mediated adenosine transient reporter for editing...

  7. Addgene's Top 10 Blog Posts of 2015

    Type
    Blog Post
    Published
    Dec. 7, 2015, 6 p.m.
    ...repository use Cas9 to cut DNA, Addgene Science Communications Intern Mary Gearing explains how Cas9 homolog,... suggestions. 8. A Match Made in Heaven: CRISPR/Cas9 and AAV Adeno-associated viral (AAV) vectors are...Validating Your Genome Edit Once you’ve used CRISPR/Cas9 to make a change to the genome, you need to check...

  8. Donations from Addgene to Yield Answers for Rare Disease Researchers

    Type
    Blog Post
    Published
    Sept. 15, 2015, 2:30 p.m.
    ...pluripotent stem cell (iPS) technology and CRISPR-Cas9 constructs from Addgene to gene-edit mutations into...looking at a bunch of different strategies with CRISPR-Cas9. There are so many different variants and strategies... of those mutations, they now plan to use CRISPR-Cas9 technology to create cell lines and/or mouse models...ve proposed to use plasmids with modified nickase Cas9 protein in order to target the specific gene region...

  9. Hot Plasmids - January 2023

    Type
    Blog Post
    Published
    Jan. 31, 2023, 2:15 p.m.
    ...targeting, with a leg up on the alternatives - RNAi and Cas13. CRISPR-Csm doesn’t degrade proximal RNAs nor does...levels of off-target effects, both of happen with Cas13. CRISPR-Csm degrades both nuclear and cytoplasmic...nuclear-RNA targeting, where siRNAs are ineffective and Cas13 has steep off-target costs. Additionally, this system...imaging tool.   Fig. 1: Csm, shRNA, and Cas13 were compared head-to-head for gene expression changes...

  10. Pushing the Limits of DNA Assembly

    Type
    Blog Post
    Published
    Sept. 6, 2017, 1:01 p.m.
    ...Producing yeast expression cassettes Traditionally, generating expression cassettes for yeast transformation...Cloning sgRNAs Production of Yeast Expression Cassettes Generating constructs with multiple point mutations...regions of homology and inserted into a linearized cassette, ready for transformation into yeast that same...

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

    Type
    Blog Post
    Published
    Aug. 20, 2024, 1:15 p.m.
    ... absence of a protein, a yes-no plot is the best-case result. For example, you have introduced a GFP protein... not be as obvious in a dot plot. If that is the case for you, switching to a contour plot should indicate...interest. Created with biorender.com.   In such cases, instead of trying to quantify based on percentage...a good statistic to compare between samples. In cases where there is no clear distinction between the ...

  12. Early Career Researcher Toolbox: Social Media for Scientists

    Type
    Blog Post
    Published
    Dec. 11, 2018, 12:46 p.m.
    ...Scientists SHOULD Be Using Twitter" podcast from Hello PhD In this podcast the hosts, Josh and Dan, discuss...monitoring that conference via social media. The podcast also features an interview with Dr. Stephani Page...and offers for postdoc positions. Check out the podcast for more tips on using Twitter as a scientist. ...

  13. How Addgene Creates Content

    Type
    Blog Post
    Published
    Nov. 17, 2022, 2:15 p.m.
    .... Expert reviews on blog posts are done on a case-by-case basis, as many of our blog contributors primarily...webinars, or use other research methods. In the case of the antibody guide, primary research and previously...changes and require updating or retiring. CRISPR/Cas9, for example, has gone through rapid change in the...

  14. Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number

    Type
    Blog Post
    Published
    Sept. 4, 2018, 12:53 p.m.
    ...positively and negatively controls the output. In this case, plasmid copy number is the input and gene expression...and keeping its expression level constant. In the case of the stabilized promoters from Segall-Shapiro ...varied in size or carried different genes. In these cases, gene expression also remained stable regardless...shuttling of the intermediates into other pathways. In cases where a specific ratio of products is needed, such...

  15. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    Published
    Jan. 14, 2016, 3:30 p.m.
    ...particular plasmid you are working with, but, in most cases at Addgene, we check what we call the “insert,” ...fusions, and multiple cloning sites. In the simplest case we have a standard backbone with one insert, and...and identity of the insert are correct. In other cases, we deal with multiple inserts or internal mutations...than two sequencing reactions. In the more complex cases, we sometimes have to design new primers to sequence...

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

    Type
    Blog Post
    Published
    Nov. 24, 2015, 3:30 p.m.
    ...experiments in specific cell populations. Listen to Our Podcast Interview with Connie Cepko Combining Cre and GFP...recombination at LoxP sites. Placing a lox-STOP-lox cassette upstream of a gene of interest blocks transcription...absent. When Cre is present, it removes the STOP cassette, activating gene expression. Although Cre isn’...versions of non-modular proteins like Cre. Could Cas9-DOG be developed soon to allow genome modification...

  17. Hot Plasmids May 2018 - Optogenetics, Decaffeination, Biosensors, and Fluorescent Protein Tools

    Type
    Blog Post
    Published
    May 15, 2018, 12:43 p.m.
    ...contributed by Brook Pyhtila  Listen to the CRY2clust podcast segment The Won Do Heo lab at the Korea Advanced...contributed by Meghan Rego  Listen to the soCoChR podcast segment Optogenetic stimulation is a powerful tool... Listen to the synthetic decaffeination operon podcast segment With the help of engineered microbes, used...FLIM-FRET acceptors  Listen to the FLIM-FRET acceptor podcast segment Article contributed by Christina Mork The...

  18. Gaining Leadership Skills Volunteering at a Professional Organization

    Type
    Blog Post
    Published
    Aug. 6, 2015, 2:30 p.m.
    ...Many thanks to our guest blogger Juliet Moncaster! Juliet Moncaster is an Assistant Professor at the Boston... post was contributed by guest blogger Juliet Moncaster Leadership skills are amongst the professional...gain such experience? Your institution may offer occasional workshops on leadership and management skills...

  19. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    Published
    Feb. 4, 2020, 2:15 p.m.
    ...unusual blue-colored protein from the jellyfish Cassiopea xamachana (Blanquet and Phelan, 1987). Eight years.... Thus chromoproteins can be used as markers in cases where cost and resources are limited or for bioart... varied greatly between chromoproteins. In some cases, cells containing high-copy plasmid-borne chromoproteins...mesogleal protein from the mangrove jellyfish Cassiopea xamachana. Marine Biology 94:423–430 . https:/...

  20. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    Published
    April 4, 2019, 12:53 p.m.
    ...antibiotic resistance selection markers, gRNAs, and dCas9 proteins. Second, it can be introduced into a broader...and a plasmid that contains the Mobile CRISPRi cassette flanked by left and right Tn7 end sequences. Donor...pneumoniae, and P. aeruginosa) in addition to Vicrio casei, a bacteria isolated from cheese. Knockdown efficiency...promoters, selection markers, and different versions of dCas9. Following integration, Mobile-CRISPRi is stably...

  21. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    Published
    Dec. 1, 2016, 3:30 p.m.
    ...systems, these systems remain valuable, especially in cases where CRISPR cannot be used (e.g. there are no suitable...targeted gene. Upon recombination, the GFP/NeoR cassette is inserted in place of exon 2. Thus, the targeted...use the Cre/Lox system described earlier. In this case, you design your targeting vector such that a set...sequence flanked by LoxP sites while the resistance cassette has been placed into an intron.    Figure...

  22. Negative Can Be Positive: Open AAV Data with Addgene

    Type
    Blog Post
    Published
    Sept. 6, 2022, 1:15 p.m.
    ...tested four serotypes containing an expression cassette with an ubiquitous promoter to express GFP in ...in a group of young rats. We found that, in this case, AAV1 (23%) and AAV6 (<5%, data not shown) were not...Addgene has shown me this was far from an isolated case. Our scientific support team answers hundreds of...But, especially for AAV data, that may not be the case. We can all acknowledge that negative data is something...

  23. What’s in Your Tube?: A Quick Guide to Measuring DNA by Spectrophotometry

    Type
    Blog Post
    Published
    July 1, 2025, 1:15 p.m.
    ...measure how light travels through a sample — in this case, the absorbance of light at three key wavelengths...residual guanidine thiocyanate. Phenol is a unique case, as pure phenol absorbs strongly at ~270 nm and ...fluorescence based assay or running an agarose gel. In the case of plasmids, the use of RNase treatment to degrade...forgotten).    Improving read accuracy    In many cases, small amounts of contaminants in your sample will...

  24. Hot Plasmids: Summer 2024

    Type
    Blog Post
    Published
    Aug. 27, 2024, 1:15 p.m.
    ... a programmable epigenetic editor consisting of dCas9 fused to a DNA methyltransferase — that can drive...DNA-targeting proteins, so the team created versions using dCas9, zinc finger proteins, and transcription activator-like...miniCRISPR” version, are much smaller proteins than SpyCas9, allowing them to be packaged in AAV vectors, potentially...histone tail (H3K4me0) to a DNA-binding protein (dCas9 shown here). D3L recruits the endogenous, autoinhibited...

  25. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    Published
    April 4, 2024, 1:15 p.m.
    ...for advice before running your analysis. In this case, I and other lab members had worked with lymphocytes... the positive one (Figure 8). Note that in many cases a stain will not result in clearly distinct positive...will show you a combination of two stains, in this case through conjugated antibodies targeting the cell...use multiple individual gates. Note that in this case, square gates were used.  Congratulations! ...

  26. Evolution of Brainbow: Using Cre-lox for Multicolor Labeling of Neurons

    Type
    Blog Post
    Published
    April 24, 2015, 2:39 p.m.
    ...translation-blocking STOP cassette immediately following the promoter. The STOP cassette includes a mutant YFP...CRISPR-Cas9 genome editing may be the hot new way to manipulate gene expression, but other gene manipulation...eliminating two color possibilities. The remaining cassette can invert as long as Cre remains expressed; once...

  27. Building Global Connections with the International Mentorship Program USA-EUROPE

    Type
    Blog Post
    Published
    Nov. 6, 2015, 3:30 p.m.
    ...Rosario F. Godino, Rocío López-Diego, & Zafira Castano Corsino. When someone says “internationalization...conceived and developed by its Director, Dr. Zafira Castano Corsino, a junior faculty member at Harvard Medical...from the International Mentorship Program Zafira Castaño Corsino is a Junior Faculty at Harvard Medical ...and mentoring activities. Follow her on twitter @zcastan or on linkedln.       Rosario Fernandez-Godino ...

  28. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    Published
    March 21, 2017, 2:30 p.m.
    ...vectors fantastic options for delivering shRNA, CRISPR/Cas9 components, and fluorescent sensors. Retroviral...allowing for selection of the transduced cells. In this case your gene and the marker can be separated by an ...rtTA) to bind TRE only in presence of Dox. In this case, your gene is repressed in steady state and is expressed...directly added to your target cells. However in that case you don’t know how many viral particles you put ...

  29. #Deckthelab Contest Winners 2017

    Type
    Blog Post
    Published
    Jan. 31, 2018, 2:44 p.m.
    ...favorite twitter #agarart accounts, Petri dish Picasso (@PetriPicasso) came together and delivered very impressive...Colorado -Anschutz Medical Campus (@CSD_CUAMC) caught a case of holiday cheer, and we loved it! Check out their...

  30. Enabling high school research at the Journal of Emerging Investigators

    Type
    Blog Post
    Published
    Sept. 5, 2018, 12:01 p.m.
    ... In this episode of the Addgene Podcast, we introduce you to the Journal of Emerging Investigators, an...interview with the Journal of Emerging Investigators Podcast breakdown Intro Chapter 1 - Introduction to Mark...the audio engineer and producer for the Addgene Podcast.      Additional Resources from the Addgene Blog...

  31. How to Deposit Your Plasmids with Addgene

    Type
    Blog Post
    Published
    Feb. 27, 2024, 2:15 p.m.
    ...a peer-reviewed manuscript OR a preprint. In the case of a preprint submission, we are happy to update...cloned into it. gRNAs are used for directing CRISPR/Cas9 to a targeted genomic location, while shRNAs are...format, e-mail us at  [email protected] Special cases Please contact us at [email protected] or +1 (...) if your deposit falls into one of the special cases below:   Pooled Library A pooled library is a ...

  32. 5 Science Rap Videos & Their Creator: Tom McFadden

    Type
    Blog Post
    Published
    Feb. 9, 2017, 3:30 p.m.
    ...with his new company, Science with Tom. In this podcast, we learn more about Tom and pick his brain for...the audio engineer and producer for the Addgene Podcast.     Additional Resources on the Addgene Blog ... a Grad Student Community Check out All of Our Podcasts Find Great Educational Content in our Plasmids...

  33. An Interview with Connie Cepko - Gene Therapy, Plasmid Tools and Insights to Success

    Type
    Blog Post
    Published
    Dec. 8, 2015, 3:30 p.m.
    ... School researcher Connie Cepko. Listen to the podcast below to hear all about the research being conducted...interview. Happy Listening! Click here to listen! Podcast breakdown: (Time : Topic) 0:00-1:00 : Introduction...the audio engineer and producer for the Addgene Podcast.     References 1.Tang, Jonathan C.Y., et al. “...

  34. Kwabena Duedu on Public Health, Biofuels, and Doing Science in Ghana

    Type
    Blog Post
    Published
    Dec. 20, 2016, 3:30 p.m.
    ...In this episode of the Addgene Podcast, we sit down with Kwabena Duedu, a researcher at the University... to Ghanaian scientists Click here to listen! Podcast breakdown (Time : Topic) 0:00 - 7:11 : Introduction...the audio engineer and producer for the Addgene Podcast.     Additional Resources on the Addgene Blog ...

  35. Antibody Validation for Flow Cytometry

    Type
    Blog Post
    Published
    Sept. 10, 2024, 1:15 p.m.
    ...endogenous levels (Laflamme et al., 2019). CRISPR-Cas9 approaches are commonly used to produce KO cell ...is not amenable to knockout approaches. In these cases, we would recommend a knockdown approach instead...lines based on expression of the target. In this case, you could compare the antibody’s labeling of the...Biddle, M. S., Smallwood, D. T., Weston, C. A., Castells, E., Bowman, V. W., McCarthy, J., Amrani, Y., ...

  36. Your Top Requested Plasmid in 2015!

    Type
    Blog Post
    Published
    Dec. 30, 2015, 3:30 p.m.
    ... lentiviral transfer vector used to deliver both Cas9 and your gRNA of interest to mammalian cells. It... blog post on Genome-wide Screening Using CRISPR/Cas9. If you need help cloning a gRNA into lentiCRISPR...References 1. Shalem, Ophir, et al. "Genome-scale CRISPR-Cas9 knockout screening in human cells." Science 343.6166...

  37. Quick, Versatile Plant Transgenesis with GreenGate Plasmids

    Type
    Blog Post
    Published
    Jan. 23, 2014, 3:23 p.m.
    ... frequently used elements in plant expression cassettes: plant promoters, N-terminal tags, coding sequences...C-terminal tags, plant terminators and resistance cassettes for selection of transgenic plants. The complete... gives the option to stack several expression cassettes onto a single T-DNA in subsequent rounds, dramatically...

  38. Searchable and Sortable gRNAs for Your Next CRISPR Experiment

    Type
    Blog Post
    Published
    May 17, 2016, 2:30 p.m.
    ...have one associated with the gRNA), application, cas9 species, PubMed ID, and depositor. The CRISPR applications...nick, purify, tag, scaffold, CRISPR-display, and dCas9-FokI. This table is both sortable and searchable...alphabetically or in ascending or descending order, in the case of numeric columns.  The data in any column are ...

  39. Hot Plasmids and Viral Preps - July 2021

    Type
    Blog Post
    Published
    July 27, 2021, 1:15 p.m.
    ...simultaneous gene activation in plants.  FnCas12a, unlike other Cas12a nucleases identified, functions at temperatures...editing in moderate thermophiles.  These evolved SpCas9 variants have improved activity on NAG PAMS and...

  40. Year of the Camelid: Antibody Style

    Type
    Blog Post
    Published
    May 9, 2024, 1:15 p.m.
    ...Camelid family is composed of camels, llamas, and alpacas (and their undomesticated counterparts) - not exactly...soon after found similar antibodies in llamas and alpacas (Figure 1). At the time, researchers were already...started with immunizing a camelid (usually llamas or alpacas). But because sdAbs are so amenable to recombinant...

  41. Oh, The Places You Can Go: Careers in Science Communication - Curriculum Development

    Type
    Blog Post
    Published
    Nov. 1, 2017, 2:04 p.m.
    ...at eScience Labs also write test question banks, case studies, and experiments for labs, in addition to...comprehensive exam proposals, a few PhD theses, and occasionally posting something science-related on Facebook...science education tools? Check out this Addgene podcast with educator and science rap mastermind Tom McFadden...

  42. Summer SciComm Series: Master of SciComm

    Type
    Blog Post
    Published
    July 21, 2022, 1:15 p.m.
    ...skill acquisition. Classes covered filmmaking, podcasting, philosophy of science, creative writing, designing...an animated video depicting nuclear fission; a podcast with a genetics researcher on parasitoid wasps;...the face of disparate spheres of research. In my case, an interest in neuroscience was carried over and...

  43. Revamp Your Lab Meetings With Creative Virtual Collaboration

    Type
    Blog Post
    Published
    Feb. 9, 2021, 2:15 p.m.
    ...unmeasurable skills and productivity coming from that casual discussion on the way to the office or in the kitchen... flexible work schedule the more we realize how casual chatter is very much missed in the virtual world...widely used and could bring back a taste of that casual kitchen chatter, ultimately sparking new ideas....

  44. Reagent Repositories Are Speeding up Science During the Pandemic

    Type
    Blog Post
    Published
    Aug. 4, 2020, 1:15 p.m.
    ...research. Image from Reproducibility for Everyone. Case study 1: Cell lines Many of the reagents used to...world shows that science is truly a global endeavor. Case study 2: Plasmids Another example of reagents that...Browse Addgene's COVID-19 plasmids and resources! Case study 3: Mouse strains Reagents developed during...

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

    Type
    Blog Post
    Published
    Dec. 10, 2019, 1:58 p.m.
    ...Th2) cells from mice that constitutively express Cas9 and screen for genes that affect Th2 differentiation...Wang Lab created a tripartite system consisting of dCas9, a gRNA fused to an RNA binding motif, and a RNA-binding...protein-bEFP fusion. Thus, the bEFPs bind the UAS via the dCas9-gRNA complex which allows transcription from the...

  46. Photosensitizer Induced Cell Ablation with FAP-TAP MG-2I-dL5**

    Type
    Blog Post
    Published
    Sept. 19, 2017, 1:20 p.m.
    ...FAP - fluorogen activating protein - dL5** in this case), and 2) a fluorogen (the TAP - targeted and activated...activated photosensitizer - MG-2I in this case). The fluorogen is not fluorescent until it binds its protein...easy to watch development in real time.  In both cases, dL5** was tagged with mCerulean3 (mCer3) for visualization...

  48. Viral Vectors 101: Viral Applications

    Type
    Blog Post
    Published
    April 13, 2023, 1:15 p.m.
    ...depending on the vector and virus. Viral CRISPR-Cas9 tools have been engineered to introduce frameshift...be used to express proteins. Viral expression cassettes typically contain a promoter (inducible or constitutive... Therapy with AAV Adenoviral Delivery of CRISPR/Cas9 Aims to Expand Genome Editing to Primary Cells ...

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

    Type
    Blog Post
    Published
    July 6, 2017, 1:59 p.m.
    ...in immunotherapy is the combination of the CRISPR/Cas9 methodology with T cell receptor engineering (Ref...using randomly integrating viral vectors, CRISPR/Cas9 genome editing allows targeted engineering of the.... "Targeting a CAR to the TRAC locus with CRISPR/Cas9 enhances tumour rejection." Nature 543.7643 (2017...

  50. How to Keep a Lab Notebook for Bioinformatic Analyses

    Type
    Blog Post
    Published
    June 2, 2016, 2:30 p.m.
    ...analysis performed in experiment 1. Use camelCase: In camelCase, each subsequent word in a file name after... more than one word should be named in the camelCaseFormat since spaces between each word can make it ...
Showing: 251 - 300 of 609 results