...from its usual location, preventing it from carrying out its proper function (Figure 3).  The Hehnly Lab...injection of plasmids and other materials.    Figure 3: Optogenetic inhibition of Rab protein vesicle trafficking...efficiency constructs are great for in vitro biocatalytic applications where toxicity doesn’t matter. Plus, for... binding of the plasma membrane. In previous publications, they described tools to control formation of.... Restricting the effect to a specific time or location can be critical when, say, the gene is essential...) and to introduce unnatural head groups for applications like fluorescent labeling via click chemistry...substantial PA accumulates in the plasma membranes, indicated by the PA probe colocalizing with SuperPLDx30....

...perform downstream characterization of, isolating 2 or 3 distinct clones to perform key experiments on is a...CRISPR-Cas System in Gene Editing. Mol Ther. 2021 Mar 3;29(3):937-948. doi: 10.1016/j.ymthe.2020.11.009  Guo C,...“Off-target effects” a vague catch-all term for unintended consequences in an experiment. For CRISPR ... example, if there are 10 putative off-target locations you can 1) determine if editing at those sites...sequencing. While this may be unnecessary for many applications, it is the only comprehensive method available...binding by Cas does not always correlate with catalytic activity at those sites, especially when DNA mismatches...For a candidate site-specific approach, mRNA quantification or western blot of putative off-targets would...

...working with RNA and extracting RNA without a kit. 3. SARS-CoV-2/COVID-19 Detection Methods Based on CRISPR...virtually. 6. Quantifying DNA? Here are Five DNA Quantification Methods to Consider by Jennifer Tsang The...The nanodrop vs. spec debate dominates DNA quantification discussions, but what about the other methods... with plasmids deposited at Addgene! 2. DNA Purification Without a Kit by Beth Kenkel To complement ...

... guest Drew Endy, in celebration of open science. 3)      Also in June, the G8 Science Ministers issued...support for open access to peer-reviewed journal publications, although many don’t necessarily publish in ...

...Blog (other programs are discussed below). Chpater 3: SITN Popularity and Events - Amy discusses some of... to be crazy complicated! Listen to the full podcast for more great science communication tips or listen...tons of ways you can get involved in science communication. In this second half of our conversation with...many things you can do start your own science communication student group and get more involved with your...is a graduate student group with the goal of communicating science to the general public and training the... the next generation of science communicators. In this chapter we learn about some of their events - science... peer-to-peer feedback in effective science communication. Chapter 7: The Goal of the SITN Podcast - Vini...

...for making scientific graphics. PS: they’re free. 3. Fluorescent Proteins 101: GFP Fusion Proteins - Making...resources can now be found in one place: the Educational Resources page. So let’s get to it. Here’s the...Anti-CRISPR, 2in1 Cloning, Fluorescent Voltage Indicators, and Photoswitchable Proteins by Cary Valley,...protein size and shape, linker sequences, and fusion location, there are many choices to make. 2. Prime Editing...Resources at Addgene Browse the repository Find educational resources at Addgene Search for plasmids at Addgene...

...that could be useful for a wide variety of reasons. 3) I would love to see more posts like this on the Addgene...Plasmids at a Low Price: Kits from Addgene Savvy Advocates Needed to Navigate a Scientific Enterprise in ... a Lab and a Project That’s it for now. As I indicated in the opening paragraph, I’m currently on my ...

...similarly but not identically to BE3, modifying a 3-5 base window 18 bases upstream of the PAM.  David...than BE2 Nishida et al., 2016 Target-AID   Edits 3-5 base window surrounding -18 position upstream of... efficiency and product purity. Science Advances, 3(8), eaao4774. https://doi.org/10.1126/sciadv.aao4774...editors catalyze purine-to-purine or pyrimidine-to-pyrimidine base changes. Solid red arrows indicate a conversion...conversion directly catalyzed by the base editor. Dotted red arrows indicate the change introduced to ... point modifications. CRISPR base editing persists through multiple cell divisions, indicating that this...target modification in primary T cells, making them a promising tool for cell therapy applications. Dual...

...Tris-acetate 40–500 kDa Tris-tricine 150 V, 1–3 hours Good for higher weight proteins Long running...very large proteins. A semi-dry transfer is faster — 3–30 minutes — and uses less buffer, but it is not as...approaches best suit your experimental needs. Table 3: Optimizing the technical design of a western blot...original publication and ensure that it includes validation data relevant to your application — and double-check...buffer, consider the subcellular location of your protein. Proteins located in membrane-bound subcompartments...or are looking to preserve specific protein modifications, we recommend doing a little more research into...Blocking solutions Your blocking options come in two categories: protein-based and chemical-based. Before selecting...

... Transcriptional repression DNA Methyltransferase 3 Alpha (DNMT3A) Vlatka Zoldoš’ lab has deposited pdCas9...available as ready-to-use lentivirus. Figure 3: dCas9-DNMT3A methylates cytosines at promoters and...epigenome (the sum of epigenetic modifications across the genome) has been implicated in disease states, and targeting...DNA sequences. Non-editing CRISPR applications direct catalytically dead dCas9 fused to a variety of epigenetic... the catalytic domain of p300 acetyltransferase increases levels of H3K27ac histone modification at specified...Preps for Epigenetic Modification Why use epigenetic modifiers? Epigenetic modification is certainly not ...Nature Communications 8: . https://doi.org/10.1038/ncomms16026 Find plasmids from this publication at Addgene...

... bar to quickly find what you’re looking for. Tip 3: Share protocols and spreadsheets with lab members...moving your paper spreadsheets for reagent storage locations, such as the list used to organize your cryo freezer...

...here for more on timely submission of a job application. 3) Read job descriptions carefully Be open minded...demonstrate in your application). Beware the myths of scientist “overqualification” but it may not be ...Utilizing existing relationships as part of the job application process can dramatically increase the chances... of a resume can be crucial for a successful application. It takes time to prepare an excellent resume...bold. You don’t have to have all of the listed qualifications to apply for a position.  A job description...target someone in a specific department and/or location. The more people you meet the better chance you...

...write our AMA submission to avoid rousing the trolls. 3. The AMA process was simple and well organized. You...re more interested in selling a product than communicating your ideas with people or discussing science...to write about in the future. The AMA was also cataloged for permanent reference by the Winnower. The only...I do a lot of public speaking where I enjoy communicating face to face with the audience. I post on LinkedIn...own expertise and your resources for science communication opportunities like an AMA. One of the best things... science communication...

..., restraining further replication.   Figure 3: Comparison of replication regulation mechanisms ...plasmids rely on their host cell's replication machinery in order to replicate—but not always to the same extent...described in our previous Origin of Replication post, DNA replication is initiated at the ori and may or...consider the replication mechanism of plasmids under stringent control, such as pSC101. DNA replication and regulation...review how E. coli normally replicate their chromosomal DNA. Chromosomal replication in bacteria is carefully...entire round of replication. Initiation is the primary control point of DNA replication in bacteria, so... the replication forks reach each other on the opposite end of the chromosome, ending replication.   ...

... leaving behind any accessory DNA sequences [2], [3]. Simply put, the piggyBac™ transposon containing ...PMID: 26702451. PubMed Central PMCID: PMC4686154. 3. A. M. Singh, D. W. Perry, V. V. A. Steffey, K. Miller...piggyBac™ technology can be used for numerous applications, including transgenesis, gene-trap screens, ...of the piggyBac™  transposase/transposon gene modification system and the scarless, Footprint-Free™ removal...technology that may be used for a variety of research applications, including making transgenic cell lines and ...PMCID: PMC2567143. 6. S. Radecke, F. Radecke, T. Cathomen, and K. Schwarz, “Zinc-finger nuclease-induced...oligodeoxynucleotides: wanted and unwanted target locus modifications.,” Mol. Ther., vol. 18, no. 4, pp. 743–53, Apr...

...variants. Klapoetke et al., Nat Methods 2014 Mar 11;(3):338-46. For more information and descriptions of...counterparts. The Janovjak lab has deposited their 3 Opto-RTK constructs – Opto-mFGFR1, Opto-hEGFR, and...turn green. Mashiko et al., Sci Rep 2013 Nov 27; 3:3355. Rinehart lab reagents for improved expression...96-102. Druckmann et al., Neuron. 2014. Feb 5;81(3):629-40.  Next-Gen Brainbow toolkit for neuronal...ligation using SpyLigase Building off their SpyTag/SpyCatcher system for protein-peptide locking, Mark Howarth... a critical role in development and are often implicated in disease progression. One of the major challenges...challenges in signaling research is the inability to replicate the spatio-temporal precision with which signaling...

...cyanobacterium Vampirovibrio chlorellavorus." PeerJ 3 (2015): e968. PubMed PMID: 26038723. PubMed Central... into a pulsating display imitating a moving caterpillar. The worm controls the snail’s behavior, forcing...exposed to predators. Hungry birds spots the “caterpillar” and then poke out the snail’s eyes and ingests...

...For treatment, it is best to identify a cocktail of 3-12 phages. Once these phages are identified, they ...researcher may only know how to prepare phage for one application but not another or they may have only used phage...Directory expands, it can also fill this role by cataloging and archiving information on individual phages...

...portion of our careers. In that time we may have only 2-3 formal supervisors that will provide guidance and ... helps you reach your goals Listens well and communicates clearly and openly Gives honest feedback without...lab. If only every science trainee had such a dedicated person guiding the start of their career. “I want...

...Qi LS, Weissman JS, Vale RD. Cell. 2014 Oct 23;159(3):635-46. doi: 10.1016/j.cell.2014.09.039. Epub 2014... its nuclease activity, catalytically dead Cas9 (dCas9) has many applications, the most well-known being...characterizing the 4D nucleome. Future modifications and applications Ma et al. developed this technique ...have major applications to cancer, for example, the visualization of chromosomal translocations or chromosomal...plasmids from this publication at Addgene. A protein-tagging system for signal amplification in gene expression...technique published every week! One of the newest applications is a colorful system that uses fluorescently...Feb 23. Pubmed. Find the plasmids from this publication at Addgene. Dynamic imaging of genomic loci...