...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...

...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...

...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...

...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...

...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...

...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:/...

...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...

...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...

...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...

... 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...

...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! ...

...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...

...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 ...

...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 ...

...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...

... 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...

...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 ...

...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...

... 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. “...

...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 ...