... plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles.  Here's ...developed an engineered adeno-associated virus (AAV) vector named AAV.CAP-Mac for non-invasive systemic gene...N.C., Goeden, N. et al. Adeno-associated viral vectors for functional intravenous gene transfer throughout...region VIII of AAV9-X1.1 to that of AAV1. This viral vector has brain endothelial cell-specific tropism, with... by pre-existing antibodies against AAV9-based vectors (and vice versa), enabling repeat dosing. In a ...

...2002). Insect cells as a factory to produce adeno-associated virus type 2 vectors. Human Gene Therapy, 13...mammalian cells, they do not require serum or an incubator to grow. The virus Insect cell systems are used...references More resources on the Addgene blog Viral Vectors 101: Producing Your rAAV AAV Titers: Where Do They...platform to produce recombinant proteins and viral vectors. Frontiers in Bioengineering and Biotechnology,...

...  Is this department/laboratory collaborative with other departments/laboratories? How long is the average...almost as daunting as the program itself. Picking a mentor comes with many challenges, and the flood of interview...more complex. “How do I reach out to potential mentors?” “Where do I start with picking universities?”...by meeting with professors - a/k/a Principle Investigators (PIs) - whose labs does research that aligns...

...microscopy, by cloning your protein of interest into a vector encoding a fluorescent tag. Once expressed as a...golgi and then transported in vesicles via the secretory pathway to their subcellular or extracellular ...bacterial components. For example, the Rainbow Vectors from the Mariette Barbier Lab can be used to fluorescently...Edition. 3.  CureFFI.org. Cell Biology 04: The Secretory Pathway. 4. Dunn, Kenneth W., Malgorzata M. Kamocka...Barbier, Mariette, and F. Heath Damron. "Rainbow Vectors for Broad-Range Bacterial Fluorescence Labeling...

...is Head of Synthetic Biology at the Sainsbury Laboratory, where she often feels more like an engineer ...the toolkit. We have included a wider range of terminators than are commonly used as we have tested them...system. The paper also gives quantitative data for regulatory elements, which has not been available for plants...easy to express genes in, which makes it a good laboratory workhorse. Tips & tricks Addgene: Do you have...start. They might want to use our promoters and terminators, but then they aren’t sure how to convert their...

...unevenly, introducing distortions in your final images. It is possible to keep distortions to a minimum with...done about that.  Second, expansion introduces distortions. The goal in expansion is for your sample to ...polymerizes, and the level of digestion. All of these factors in turn depend upon the specific gel recipe used... L (2022) Super-resolution microscopy: a brief history and new avenues. Philosophical Transactions Royal...

...you possess because any potential employers, collaborators, or co-workers want to know that you are a well-rounded...situation? Maybe you demonstrated empathy for a collaborator or co-worker and showed that you really understood... for working with lots of students, managing inventories in the lab, or organizing a research project,...Resources Cross team communication  External collaborators For further advice on highlighting your transferable... for non-academic science careers Learn about mentoring Resources on Addgene.org Check out our careers...

...2020). In the two-plasmid system, an effector plasmid (pEffector) expresses all seven required genes and...CRISPR-Cas systems, in contrast to single-protein effectors employed by Type II systems (such as CRISPR-Cas9...plasmid (pDonor) encodes the mini-transposon. pEffector and pDonor can also be combined in single-plasmid...technologies. Message or follow him on twitter @Vopinator.     References: Halpin-Healy TS, Klompe SE, Sternberg...

...CRISPR/Cas9 Synergistic Activation Mediator) uses an improved activator design that consists of inactive... domains that recruit additional transcription factors. The Root and Doench labs utilized tracrRNAs with...stem loops to recruit additional transcription factors to the targeted region (Sanson et al., 2018) . ...tracrRNA was combined with the sgRNA expression vector allowing the screen to be conducted with only two...

...introduce any one of these topics without some editorializing, and he wisely makes no such attempt. He is...discusses have been making the rounds in journal editorials, science social media, and conference workshops...biology also falls within this section. The GFP story has its own Franklin in Doug Prasher, who originally...describes the detection of gravitational waves--a story I was much less familiar with as a biologist--as...12 years, I’m very familiar with the role our repository has played in these fields, and seeing names ...

... in Parkinson’s disease. Knowing the primary investigator where the plasmid was created can narrow your...but I can’t browse through 300+ plasmids. Under ‘Vector Types’ I selected ‘Lentiviral’ which eliminates... coveted Blue Flame is more than an award for depositors and inspiration for our updated mascot, it’s ...model system for your research (e.g. Mammalian) Vector Type: What type of experiment are you planning?...Identify a specific marker protein Use: Similar to “Vector Type” you can filter by experimental purpose (e.g...

...acids derived from the crystal jellyfish Aequorea victoria. It has a fluorescent emission wavelength in the... forms spontaneously and without additional co-factors, substrates, or enzymatic activity – it only requires...that the protein could be taken directly from A. Victoria and expressed in any organism while still maintaining...promoter of interest can be used to effectively monitor gene expression from that promoter in a given cell...optogenetics at OpenOptogenetics and find optogeneic actuators and sensors at Addgene. Cell marking/selection...

...on the Society for Laboratory Automation and Screening (SLAS) Electronic Laboratory Neighborhood.   Work...And, yes, your partner CAN take the kids to the doctor without your help. Your partner is equally responsible...always get done, but you’ll have time to read a story to your baby or run that extra two miles which is...eight-year-old daughter vehemently reminded her doctor that she was allergic to penicillin – I am much...

...much information as possible from depositing laboratories and associated publications, but our quality...Generation Sequencing (NGS) technology. As our repository continues to expand, we realized the need to ...user interface, so our QC team could initiate and monitor the entire pipeline. Benefits of Nextflow for ...user interface. In our next post, we’ll share our story about how we used Nextflow Tower to augment our ...Keep a Lab Notebook for Bioinformatics AnalysesAAV Vector Quality Control: Going the Extra Mile with NGS ...

... plasmids, antibodies, and viral preps in the repository through our Hot Plasmids articles.  Here's ...requiring either expressing it with additional cofactors or using significantly truncated forms that limit...more feasible, the lab of Alessio Ciulli and collaborators developed CRBNmidi (Kroupova et al., 2024). ...abscessus, an increasingly common human pathogen notorious for its high resistance to antibiotics. Several...Bentley Time to get flexible! Alex Dunn’s lab and collaborators have deposited a new mechanosensor called PILATeS...

...genetically encoded fluorescent probe. Several factors contribute to the popularity of GFP including (...organisms and cell types, (ii) no need to add a co-factor, (iii) easy visualization with standard filter ..., proteins that bind endogenous fluorescent co-factors can be used. Examples include the LOV domains that...Kumagai, 2013). Next to these naturally occurring co-factor binding proteins, systems wherein a protein binds...Smaller genetically encoded probes are based on cofactor binding proteins as described above. The smallest...

...to capture ideas, write down certain notes or brainstorm, but we have to admit that paper-based record...means to manage their work, combining data, team, inventory and project management functionalities with experiment... and included hundreds of researchers. The full story is available here. Our questions focused on researchers...all types of data, which enables researchers to store everything in one place. From pictures to excel ...lab (user, date, time, action) are required for regulatory purposes in many labs. Besides this being of ...

...motifs within transmembrane proteins, such as receptors, or post-translational modifications on those ...) cells. Typically, you would label the CD8 co-receptor protein on the surface of the CD8+ T cells and...cytosolic proteins, as well as proteins of the secretory pathway. Those can include cytokines and chemokines...Examples of such targets include transcription factors, histones, and DNA repair enzymes. Note that when.... His projects include investigating negative regulators of T cells and cytokine signaling in tumor immunity...

... 37°C heat block, LED blue light illuminator or UV light illuminator. Approach: Highly similar to the ...equipment, Fluorescent emission reader Approach: FnCas9 Editor Linked Uniform Detection Assay (FELUDA) is a technique...method based on CRISPR-Cas12. Cold Spring Harbor Laboratory. https://doi.org/10.1101/2020.02.29.971127 Rauch...SARS-CoV-2 Genetic Material. Cold Spring Harbor Laboratory. https://doi.org/10.1101/2020.04.20.052159 Zhang...

...editing approaches for exon skipping. Addgene depositor Charles Gersbach used paired zinc finger nucleases...multiple exons that, when deleted through NHEJ, will restore the dystrophin reading frame. gRNAs bind in intronic...this result, Nelson et al. find that dystrophin restoration is maintained for at least six months. Future...stem cells 2. Remove any immunogenicity of the AAV vector Once delivery has been optimized, it’s important..., Olson EN. Postnatal genome editing partially restores dystrophin expression in a mouse model of muscular...