We narrowed to 1,010 results for: PLE
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TypeBlog Post... channels and pumps in developmental biology. Examples of some ion-channel and ion-pump encoding genes...genes HCN1 KCNJ2 KCNJ13 GABRG1 KCNIP2 KCNQ1 Examples of some genes encoding gap junction proteins cx43...also modulating bioelectric states in vivo. For example, the Levin lab from Tufts University has used Arch...sensitive fluorescent voltage indicator with near-complete desensitization in continuous light. ArcLight ...
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Hot Plasmids - March 2019 - Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins
TypeBlog Post...minutes of inactivating the CASANOVA complex, fluorescent CRISPR complexes were observed binding telomeric ...ever like to write about a recent plasmid deposit please sign up here. Listen to this episode ...2015) or ratiometric Bimolecular Fluorescence Complementation (rBiFC, Grefen and Blatt, 2012). The 2in1 Plasmid... -
Reproducibility for Everyone: Lessons from an Open Science Collaboration
TypeBlog Post...accessible, anyone could present the material. For example, Addgene’s contributions by Tyler Ford included... and matched to serve a specific audience. For example, I joined this effort later in the year and in ...diverse and international team assembled the right people at the right time. Be inclusive and seek solutions...like to join or contribute to this collaboration, please check out the official website, follow the group... -
Celebrating 15 Years of Scientific Sharing
TypeBlog Post...service, and more For the first time in the last couple of years, we don’t have to worry about having enough...community is loving it. Almost 10,000 viral vector samples were distributed in 2018 and we are poised for ...unsolicited praise: “Addgene, for one, is a sterling example of collaboration and innovation between legal and...Our long reaching reputation helps us hire great people from farther away via connections and even via ... -
Stabilized Bacterial Promoters: Constant Gene Expression at any Copy Number
TypeBlog Post...promoter region, which is predicted to be required to decouple gene expression and plasmid number. In a suite...fluctuations. When cells are dividing rapidly, multiple rounds of replication are initiated before the...greatly impacting the level of gene expression. For example, the synthesis of deoxychromoviridans requires ...products is needed, such as within a multi-protein complex, stabilized promoters can eliminate variability... -
Hot Plasmids April 2018 - Protein Degradation, Nanoscopy, FIRE-Cas9, and Yeast Expression Tools
TypeBlog Post...ever like to write about a recent plasmid deposit please sign up here. No time to read? Listen to our Hot...aberrantly activate the proteasome. Clift et al. tested multiple methods for TRIM-Away delivery. For small cell...rapid and reversible recruitment of chromatin complexes to any genomic locus in any cell type. By varying...FIRE-Cas9, experiments can be designed to target multiple loci at once. In addition, the approach allows... -
The Future of Research Symposium Boston 2015
TypeBlog Post...on the symposium as a whole and for registration please go to http://futureofresearch.org/boston/. The ... about how the system operates and functions” “Complete lack of information on number of postdocs” “Postdocs...comments are well correlated with published data. For example, in 2010, less than 15% of postdocs had achieved...beginning to address these issues independently; for example, Harvard’s Therapeutics Graduate Program has a ... -
Progress Towards a PAM-Free CRISPR
TypeBlog Post...enzymes The solution to the PAM problem seems simple, in theory: find a Cas enzyme or evolve one that...Cas proteins that don’t utilize NGG There are plenty of Cas9 variants which utilize alternative PAMs...engineering of the PAM-interacting domain of Cas9. After multiple rounds of engineering, they developed the SpRY...display higher off-target effects than SpCas9, a pleasant surprise since the PAM flexibility could in theory... -
Plasmid-based Recombinant Monoclonal Antibodies: What They Are and Why You Should Be Excited About Them
TypeBlog Post... around the world to detect, purify, quantify, deplete, and visualize proteins of interest (Greenfield...antibody expression. If starting from a protein sample (for instance, affinity purified antibodies from...recombinant monoclonal antibodies for enhanced multiplex immunolabeling of brain. eLife 8: . https://doi.org...NH, Rahman MZ, He L, Xin L, Shan B, Li M (2016) Complete De Novo Assembly of Monoclonal Antibody Sequences... -
Meet an Addgenie: Insights into Grad School, Career Paths, and More
TypeBlog Post...myself. I like being with the people and for the people, feeding the people, and making them get outside...enjoy the field I am in, the atmosphere, and the people.” My absolute favorite response to this question... -
BeHeard Award 2018: Diseases of Glycosylation, Arginine Mutagenesis, & Neural Development
TypeBlog Post... into functional multi-subunit macromolecular complexes, but a serendipitous discovery led them to investigate...different Arg-mutant enzymes could be rescued by supplementation with guanidine hydrochloride (GdnHCl). Rescue...previous approval by the FDA, GdnHCl could, in principle, provide an effective treatment for rare diseases...concentrations (≤3 mM) of GdnHCl can functionally rescue multiple Arg-mutant proteins in living baker’s yeast cells... -
Rabies and Neuronal Tracing
TypeBlog Post...EnvA receptor (TVA) in user-defined cells. For example, a researcher can deliver Cre-dependent TVA to ...optogenetic or chemogenetic receptors) enables complex control of specifically defined neuronal populations...how newly generated neurons integrate into the complex pre-existing circuit of the brain. Another study...circuitry, and continue to contribute answers to the complex questions of how the brain operates. References... -
The Materials Science of Optogenetics Experiments
TypeBlog Post...different activation/inactivation kinetics. For example, the light-gated cation channel, Channelrhodopsin...Therefore, light must be delivered in pulses. An example of a standard stimulation parameter is 20Hz (laser... control of motor coordination by Gi/o protein-coupled vertebrate rhodopsin in cerebellar Purkinje cells... Central PMCID: PMC3769938. 5. Iwai Y, et al. A simple head-mountable LED device for chronic stimulation... -
Magnetic Control of Proteins: More than a Dream
TypeBlog Post...fluorescence when a small magnet was passed over the sample (Frank Hayward et al., 2024). The fluorescence ...intensity decreased whenever the magnet approached the sample (by less than 1%) and recovered after the magnet...interference, but the result persisted. (Even better, multiple other labs have reproduced their EGFP findings...expressing MagLOV while a magnet is waved under the sample. The imaged area measures approximately 82 mm across... -
PhD Applications After COVID
TypeBlog Post...where the PhD application cycle becomes much more complex. “How do I reach out to potential mentors?” “Where...of schools by meeting with professors - a/k/a Principle Investigators (PIs) - whose labs does research...right fit. Reaching out to PIs also means doing plenty of mini-interviews; these professors will not only...campus even if you can’t visit. By talking with people at the institution, you can get a better sense ... -
xCas9: Engineering a CRISPR Variant with PAM Flexibility
TypeBlog Post...alternative PAM sequences are longer and often more complex than NGG, resulting in lower overall genome frequency...According to this logic, ω-dCas9 variants recognizing multiple PAMs should have a fitness advantage over proteins... and CAA PAMs, they saw that xCas9 3.7 cleaves multiple PAMs at a higher efficiency than SpCas9: NGT (...modifications that can be made to Cas9. It is a pleasant surprise that selecting for one beneficial trait... -
Fluorescent Tagging of Endogenous Genes with SapTrap
TypeBlog Post...site destination vectors for inserting a tag at multiple target sites. SapTrap vectors can be easily modified...Cas systems used for endogenous protein tagging, please visit our website CRISPR/Cas Plasmids - Protein... assay additional protein characteristics. For example, Masato Kanemaki's lab has developed a CRISPR/Cas-based...PMC4974478. Natsume, Toyoaki, et al. “Rapid Protein Depletion in Human Cells by Auxin-Inducible Degron Tagging... -
Viral Vectors 101: Voltage Indicators
TypeBlog Post...signals are integral to many facets of biology. For example, many heart issues stem from the heart beating ...which are electrical charges propagated through multiple cells to do things like translate perceived signals...naturally occurring voltage indicator. To give an example, a genetically encoded phosphatase (voltage sensing...then localize and emit signal from. This specific example is attractive due to the large number of sheer ... -
Plasmids 101: Choosing an Antibiotic Resistance Gene
TypeBlog Post...selecting plasmid-containing bacteria. So… why do some people still use amp? Carb is expensive (relative to amp...host/plasmid DNA during selection if it’s not completely bound up. While this likely occurs at a low rate...attention to antibiotic selection. If you feel like multiple of the above antibiotics would work for you, they...Antibiotic Resistance Stability Time Saving? Works for multiple organisms? Compatible with most bacteria? No satellite... -
15 Hot Plasmids from 2017
TypeBlog Post...way to increase CRISPR editing efficiency across multiple CRISPR enzymes. Plasmids for CRISPR coselection...was also independent of another TTAGGG binding complex, shelterin. Swapping the DNA binding domain of ...segment on regulating telomere length with TZAP Multiplexible plant genome engineering toolkit The Voytas ... well as Cys4, tRNA, and ribozyme systems for multiplex gRNA expression. Expression of toolkit components...that are sure to be enabled by this toolkit. For examples of the toolkit in action, see Cermak T, et al....