...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...

...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...

...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...