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We narrowed to 716 results for: cas

Showing: 361 - 380 of 716 results

  1. How Addgene Creates Content

    Type
    Blog Post
    .... 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...

  2. Early Career Researcher Toolbox: Social Media for Scientists

    Type
    Blog Post
    ...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. ...

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

    Type
    Blog Post
    ...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...

  4. Addgene's Tips for Plasmid Quality Control

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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...

  7. Gaining Leadership Skills Volunteering at a Professional Organization

    Type
    Blog Post
    ...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...

  8. Mobile-CRISPRi: Bringing CRISPRi to Diverse Bacteria

    Type
    Blog Post
    ...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...

  9. Chromoproteins: Colorful Proteins For Molecular Biology Experiments

    Type
    Blog Post
    ...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:/...

  10. Plasmids 101: Knockout/Knock-In Plasmids

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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...

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

    Type
    Blog Post
    ...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...

  13. Hot Plasmids: Summer 2024

    Type
    Blog Post
    ... 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...

  14. Antibodies 101: Introduction to Gating in Flow Cytometry

    Type
    Blog Post
    ...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! ...

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

    Type
    Blog Post
    ...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...

  16. Adenovirus Guide

    Type
    Guide
    ...gene expression cassettes are often too big to be packaged in rAAV vectors. This is the case of some commonly...capacity is large enough to carry the Cas9 gene and a gRNA expression cassette in one single viral particle. ...production of replication-competent adenoviruses (RCAs) that would be able to infect cells and replicate...DNA. However, if the plasmid contains a Gateway cassette containing the ccdB gene, a ccdB-resistant strain... or when needing to package larger cargo. CRISPR-Cas9 Delivery As compared to other viral vectors, rAdV...commonly used Cas9 homologs, especially when having to express them alongside a guide RNA and other regulatory...exposed. There is a risk of spontaneous reversion to RCAs, although this has been reduced by providing the...

  17. Adeno-associated virus (AAV) Guide

    Type
    Guide
    ...Addgene. CRISPR/Cas9 Delivery rAAV vectors are a popular choice for the delivery of CRISPR/Cas9 elements, making... genome editing. Packaging the Cas9 from Streptococcus pyogenes (SpCas9) and a gRNA together into an rAAV...this obstacle, has been to use smaller Cas9 orthologs, such as SaCas9 from Staphylococcus aureus . The Feng...regions between the two ITRs with a gene expression cassette that typically contains the transgene of interest...DNA. However, if the plasmid contains a Gateway cassette containing the ccdB gene, a ccdB-resistant strain...Feng Zhang lab has developed plasmids containing SaCas9 optimized for their use in rAAV vectors . Read ...too large to fit within the rAAV capsid, such as Cas9-derived plasmids and base/prime editors, is to create...

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

    Type
    Blog Post
    ...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 ...

  19. Quick Guide to All Things Lentivirus

    Type
    Blog Post
    ...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 ...

  20. #Deckthelab Contest Winners 2017

    Type
    Blog Post
    ...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...
Showing: 361 - 380 of 716 results