We narrowed to 842 results for: Eras

... strongest interaction between the guide and the target and 0 being the weakest interaction due to mismatches...such that weak interactions (e.g., scores <20) are reduced to 0 and strong interactions (e.g., scores ... Zuckerberg Initiative. We hereby announce the general availability of new a tool for CRISPR scientists...project is funded by CZI and the CZ Biohub for the acceleration of science. It's open-source. You can see all...analyses, but it also allows you to download any generated info for your own purposes, or delete the originals...that it can be used in the rank equation above. Generally, the greater this distance between the cut and...sequence inserted by HDR. Although this practice is generally advised, there is little detail published on how...

...sequencing (HTS) data we generate and maintain. It became clear that while several people across teams had...enjoy is discussion of current new and exciting literature. As I transitioned out of academia and away from...researchers of all varieties must stay current with literature and emerging technologies to remain competitive...Microbial Pathogenesis that occurred once a week for several months at a time during the spring and fall semesters...club to begin! Figure 1: Discussing next-generation sequencing at the Addgene bioinformatics journal...teams or to stay abreast of current trends in the literature, sampling a breadth of work from a field (perhaps...wandering monologue?  Assuming the feedback is generally positive and the meeting seemed to be of use, ...

...droplet quality. Using a droplet generator, the sample and the droplet generation oil are moved through small...within the expected range. We have also found that generating the standard curve for qPCR can be challenging...Pipette slowly If you are using a manual droplet generator, you will have to be careful about transferring...transferring your droplets from the droplet generator to the PCR plate. Maintaining a high droplet count is important...sample. Even if you are using an automated droplet generator, you should avoid pipetting too quickly when making...rate of 2C/s is recommended to ensure an even temperature change among all of the droplets, but you can...recombinant adeno-associated viral vectors." Human Gene Therapy ja (2019). PubMed PMID: 31140327. PubMed Central...

... Adenovirus Vectors for Gene Therapy, Vaccination and Cancer Gene Therapy. Curr Gene Ther, 13(6), 421–... anecdote goes toward saying that host-virus interactions are an important part of the biology of adenoviruses... the commercial market). Although host-virus interactions likely seem important for any virus, we will...initiating the whole process – such as the viral DNA polymerase – it makes sense that vectorization strategies...oncolytic virus in addition to standard uses in gene therapy and in basic research, like other viral vectors...PMID: 31749409. Harrach, B. (2014). Adenoviruses: General Features. In Reference Module in Biomedical Sciences...novo viral gene expression: Implications for gene therapy. Proc Natl Acad Sci U S A, 95(19), 11377. https...

...disease. This work will hopefully accelerate the development of targeted therapy for peritoneal mesothelioma...learn how plasmids from Addgene will be used to accelerate rare disease research and provide hope to patients...which disruption of the smn1 gene results in the degeneration of spinal cord motor neurons. This disease is...using C. elegans as a model to identify SMN1 interacting partners and perform whole animal screens for...for small molecules that block the neurodegenerative effects of smn1 disruption. As part of his work in...will use Addgene plasmids to characterize smn1 interactions with other genes. He will use fluorescent proteins...of the molecular biology behind SMN1 and its interaction partners, Dr. Di Schiavi will be able to better...

...markers all tagged with mEmerald. Plasmids to monitor protein-protein interactions through multicolor labeling...scale: Look at the mEmerald variants first, followed by the color variants. The mEmerald plasmids offer a...biology, seeing what interacts with what, what affects what. It's like fixing a Maserati. If we know how all...Fusion Proteins - Making the Right Connection.  mEmerald plasmids and many more! Baird has a helpful tip...the Davidson lab always tagged new proteins with mEmerald first, Baird says. From F-Actin to mitochondria... memerald...

... Choosing a Cas9 for your knockout experiment Generating a no-fuss genetic knockout  Knock-out cells or...loads of sequences to select from - in many cases several exons’ worth of DNA. Any ∼20 nucleotides of DNA...targeting opposite strands must be available in moderate proximity to each other.  If having two sgRNAs...requirements, giving it more flexibility. A consideration with this tool is that the desired edit will...strand break.   Prime editors are the second generation of base modifiers available. This system utilizes...transcriptional activation domain - VP64.  Later generations of activators include the SunTag system which... The Synergistic Activation Mediator complex, generally the strongest activator, consists of a dCas9-VP64...

...and discovery The first generation of CRISPR could cut DNA, the second generation could do base editing,...specificity: when purified, it could be programmed to generate specific cuts on the RNA target. In addition, ...most common RNA targeting system, Cas13, the "collateral" activity of the enzyme can lead to off targets...RNA editing tools.   Fig. 1 Schematic of luciferase RNA knockdown assays using Cas7-11   Use ...editing, and with the third generation came Prime editing — the ability to now make any small changes, including...small deletions and small insertions, leaving a several kilobase-sized gap in the gene editing field. The...precision into a genome, can be considered the fourth generation of CRISPR, one which drops itself neatly into...

...and the moderators were on top of any comments that were at all trollish. Kudos to the moderators for keeping...lists of forums on particular topics. Volunteer moderators set the rules for these forums and try to keep...questions! 2. The r/Science subreddit is well moderated. One of our original worries about sharing science...trolls (urban dictionary for trolling: The art of deliberately, cleverly, and secretly pissing people off, ...dialogue). Luckily r/Science has a great team of moderators who facilitated scheduling, filtered out inflammatory... sales-y in posts which falls in line with our general mission to share- as opposed to sell- science. ...expect. Fortunately, Reddit Science is heavily moderated. I started watching the questions and comments...

...practiced or required by many other journals and several funding agencies (e.g. Wellcome Trust, NIH Data...information (or links to information) for assessment of veracity of the conclusions drawn. The main difference ...publication. Who is to say that the initial data-generating lab has efficiently trawled their own data? A...much time they had. It could be argued that data generators may, in light of the policy, hold onto data longer... available to everyone at once, including the generators. This practice doesn’t appear to harm the originating... as in an actual publication and not require interaction with the data authors. In some types of research...multiple animals on different occasions via video, generating huge amounts of data. When published, this data...

...(2023) Nanobodies: A Review of Generation, Diagnostics and Therapeutics. Int J Mol Sci 24:5994. https:...The UN General Assembly has declared 2024 the International Year of Camelids. The declaration is intended...into the clinic as well. The first sdAb based therapeutic was approved in 2018, with many others in clinical...clinical trials (Jin, et al. 2023). In addition to therapeutics, there is a lot of excitement around using sdAbs... synthetic single-domain antibody libraries to generate new sdAbs. There has also been some interesting... (A) Overview of genetic modifications made to generate the nanomouse. (VHH = variable domains of heavy...their impact grow as new sdAbs become easier to generate and more widely adopted. So thank you to the camelids...

... due to the interaction with quizzes and discussion on that week’s topic by the moderators. The assignments...scientists. Over the course of the class, the moderator discussed individual sections of a paper, the ...laborious, but helped build my understanding of how to generate a manuscript. At the end of the online course,...understanding of how to apply it. The personal interaction of critiquing others’ manuscripts as well as ...the writer works.  4. Practice writing everyday Overall, the most important instruction the classes taught.... Over time that portion of the paper will be generated. After you complete a publication quality figure...figure. Writing a scientific manuscript is like generating a quilt with your lab mates, you add the individual...

...Addgene blog Important Considerations When Using AAVs Three Key Considerations For Precise Neuronal Targeting... for sharing on the site. The entry form has 5 general question types and it takes approximately 15 minutes...experiment, read our blog post on important considerations when using AAVs. Experimental results: These...virus? If you were using a genetic tool, did it operate as expected? If your experiment did not work, we...Frontiers in Neuroanatomy and summarized some considerations for precise neuronal targeting using AAV on...submission of an experimental image result that you generated with a caption. For microscopy images, please ...microscope used, and objective used. If you did not generate the image yourself and it is under copyright, ...

...high efficiency CRISPR-based approaches for the generation of knockins via homologous recombination with...approach is most well suited, since it routinely generates knockin efficiencies of 1-40% with G418-selected... CRISPR-based approach. Second, the AAV preps generally used for gene editing are low titre, and are therefore... most current CRISPR-based approaches for the generation of knockins, it is not always ideal. Because ...advantages of the AAV and CRISPR-based approaches, several groups have recently worked to enhance the efficiency...AAV and CRISPR-based approaches is described in several recent reviews (Howes and Schofield, 2015; Moser...PMC3010411. 2. Kohli, Manu, et al. "Facile methods for generating human somatic cell gene knockouts using recombinant...

...kits with greatly reduced costs per plasmid. We generally create kits to allow you to easily use a set of... by making it easy for a researcher to quickly generate N- and C-terminal epitope tag fusions to a protein...protein of interest. The plasmids used to generate these fusions all use the same restriction enzymes for...provided a solution for this problem by developing a general translation initiation sequence known as the Species-Independent...locus-specific libraries compatible with Illumina next generation sequencing in order to study evolutionary dynamics...These bridging primers are amplified and used to generate Illumina sequencing libraries using a 2-step PCR-based...open source platform and the libraries can be generated in a cost-effective manner with minimal bias. ...

...Standard AAV vectors are generated with a producer cell line like the widely used 293T. During vector...Casey Maguire and colleagues from Massachusetts General Hospital and Harvard Medical School took a closer... antibodies. This is particularly useful for therapeutic applications in vivo where endogenous anti-AAV...anti-AAV antibodies often neutralize the therapeutic efficacy of AAV vectors. How are exosome AAVs produced...exosomes or stabilize their structure or their interaction with AAVs (Schiller et al., 2018). Exosomal vesicles...interest special contexts, such as the delivery of therapeutic proteins to the brain which requires crossing...cells using exosome-associated AAV." Molecular Therapy 25.2 (2017): 379-391. PubMed PMID: 28082074. PubMed...

...and easily generate zebrafish transgenic lines to study neurobiology and neuorodgenerative diseases. The...Gateway-Compatible Toolbox is based on the original Tol2kit generated by the Chi-Bin Chien lab (Kwan et al., 2007) and... specific neuronal cell types involved in neurodegenerative diseases, the Cole Lab selected promoters ...The impact of KLF4 N-terminal variants on iPSC generation Deriving iPSCs (induced pluripotent stem cells...reprogramming somatic cells. A common method for generating iPSCs to express the transcription factors from...Polycistronic cassettes that contained KLF4s had overall diminished KLF4 expression levels; this altered...mScarlet. Bindels et al used a combination of deliberate and random mutagenesis to isolate the best RFP...

...(tracrRNA) which interacts with the Cas9 protein. These RNA molecules are vulnerable to attack by both...effectively serving their function of guiding Cas9 to generate a targeted break. These advances in gRNA technology...foreign RNA. To protect the RNA from degradation, several simple methods have been employed which modify ...modification is best? Studies suggest coupling several of them together, for example phosphorothioates...more efficient on a budget is to simply swap out several of the ribonucleotides for deoxynucleotides. A ...A partially DNA gRNA is surprisingly well-tolerated by the Cas9 protein and can increase on-target efficiency...photocleavable gRNAs. Photocleavable guides are generated by introduction of a single photocleavable 2-nitrobenzyl...

...neurons and astrocytes By Mike Lacy Cell-cell interactions underlie many kinds of tissue function, but ...To better understand how astrocytes and neurons interact, the labs of Baljit Khakh and colleagues mapped... The team expressed extracellularly-anchored horseradish peroxidase (HRP) in striatal astrocytes and neurons...studies of the molecular bases of astrocyte-neuron interactions. Plus, the tools used here (also available as...molecular foundations of their multicellular interactions. Neuron, 113(16), 2599–2620.e7. https://doi....peptide that caused nucleolar disruption into a general "killswitch" for other biomolecular condensates...this perturbation interrupted the functions of several endogenous or pathological condensates, including...

...Promoters control the binding of RNA polymerase and transcription factors. Since the promoter region ...protein, the repressor protein can bind to the operator. The bound repressor then prevents transcription... repressor (TetR) can bind to the tetracycline operator sequences (TetO), preventing transcription. In...have been engineered and improved. Takara’s “2nd generation Tet-Off vectors (Tet-Advanced)” are the most ...VP16. cTA binds to promoters containing putative operator sequences (CuO) (linked in groups of 6), allowing...the same manner, with LexA binding to the lexA operator (lexAop). Fusing the LexA DNA-binding domain (...refined the Q system and introduced the second generation QF2 activators. These activators solve the problem...