...very high level of independence, such as museums, YouTube channels and podcasts, science pub nights, and...plumber, a physicist, an ambassador to Spain, and a high school student would all be defined as laypeople...comes to molecular biology.  Some lay people have a high amount of knowledge or interest in STEM subjects...it would be safe to assume that they both have a higher level of science knowledge than average and are...scientist underrepresented minorities in your local high school who are also students in AP biology classes...

...well-organized, documented, and easy to find (though we might need to see it to believe it.) As you browse the...epitope that was previously hidden, an antibody might suddenly work that didn't before. This is most often...antibody to bind to a previously hidden epitope (right). Created withBioRender.com.   Biological Processes...it is attached to a cell membrane.  Picking the Right Antibody  It can get even more complex if you’re...configuration. So how do you ensure that you have the right antibody for the protein as it exists in your assay...how the available epitopes in any given protein might change depending on the conditions and the assay...

...drastic shift won’t happen overnight, but there are a few reasons why you might choose NgAgo over CRISPR...TtAgo-mediated cleavage in mammalian cells, likely due to high temperature requirements. To solve this problem,...preclude future HDR, which could lower HDR efficiency. Higher sensitivity to off-targets More research is needed...other small nuclear DNAs from binding NgAgo, it might make it difficult to use NgAgo to disrupt multiple...Addgene Blog Read About Cas9 Homolog Cpf1 Check Out High Fidelty Cas9 Mutants Browse All Genome Engineering...

...Fighting with antibodies to produce immunohistochemistry images that are crisp, bright, and lacking in...find high levels of background staining by the secondary, then the following three strategies might help...strategies for mitigating this issue, and we’ve highlighted a few below.  What’s the problem?   First, why...

...explains, while these Twitch sensors are always bright, they change color as they shift from "off" to ...They identified a toadfish TnC variant which had a high-affinity for calcium binding and developed a version...see tonic action potential firing in neurons and high resolution functional tracking of T lymphocytes,...and acceptor domains. He says, these new colors might now enable observation of the interaction between...Addgene? Check out our Hot Plasmid page, where we highlight some of the popular & newly available plasmids...

... sections of scientific articles (gasp). To my delight, not only was I able to make the edits without ...all about the MOI. In retrospect I used a way too high Cas9 MOI considering that A549 cells are relatively...robust cell line. If your cells are sensitive, it might be more difficult to get them to grow out. The only...presence of the band, you could run another gel that highlights the important results (for example, if you needed...a human cell line with a gene knockout was straightforward, and is a testament to how far genome engineering...David Root and is described in Rational design of highly active sgRNAs for CRISPR-Cas9-mediated gene inactivation...

...butterflies in your stomach, the virtual approach might be less stressful. For both conferences, there was...Bachle. (For presenters, there’s also less stage fright!)And depending on the conference, the audience ...can upvote a question and the host will see the highly voted questions,” says Zhang. “Usually the moderator...moderator starts by asking the speaker the highly upvoted question.”If the Q&A session ran out of time, ...for Everyone.  While this type of interaction might not work when you are giving a research talk, there...difficult in the virtual format than in-person. You might miss out on the short chats between sessions or ...

...reporter that has a large dynamic range and high brightness. The Shu Lab from UCSF designed an improved...reporter that has a large dynamic range and high brightness. This reporter achieves approximately 100-fold...immune system, and cell turnover. Apoptosis is a highly controlled process that is triggered by internal...a TEV protease cleavage site. TEV protease is a highly specific protease that is commonly used to cleave...

...neuroscience novice (newb). In this post, I’d like to highlight some of the things that popped out to me as a ...novice and indicate why other neuroscience newbies might want to get into the field. Find Optogenetics Plasmids... neuroscience but are afraid you don’t have the right kind of biological expertise to get into the field...’ve developed in your current work to make new insights in neuroscience. Even if you're currently a synthetic...interested in using AAVs because they provide a highly versatile means of delivering genes to many different...

...potential for microbial contamination. Additionally, the high activity of these enzymes implies that even a minute...a protein of interest). Mesophilic microbes are highly preferable (and successful) as host systems since...coli expressing the chaperones protein resulted in higher levels of growth compared to the E. coli strain...an outreach scientist from Addgene. The talk highlighted the importance of reagent repositories and the...leveraged for harmonious progress.     We’re always delighted to engage with potential collaborators and interested...

...match your needs. If YFG is thrillingly new, you might have to make a transgenic fly in order to study ...study it. The process is fairly straightforward. Basically, you place your gene of interest in a plasmid and... trying to overexpress or knockdown a gene, you might need different plasmids. Harvard’s DRSC/TRiP Functional...gene to select for transformed bacteria, but you might be wondering, “How do I know which of my flies have...integration will have eyes that are anywhere from light yellow to red. The variation in color depends on...strategy and the services offered by the company, you might have to map into which chromosome the transgene ...

...is an extremely useful technique that provides insight into protein:DNA interactions. ChIP works by using...exploring these interactions, and can give valuable insight into DNA expression and protein interactions, including...require crosslinking, which uses formaldehyde or UV light to create strong but reversible bonds. This will...options for this step are sonication, which uses high-frequency sound waves to break up the DNA, or enzymatic...attach the proteins to the DNA, depending on how tightly the proteins bind natively. But the amount of cross-linking...few rounds of adjustments, to get everything just right!  Fig. 2: Goldilocks is actually celebrating...

... activation and knew she was on the right track. The next insight was to assemble a synthetic transcription...bioengineering expertise? “This is the next step in high-throughput, genome-wide screening,” Konermann said...sgRNA library. Our SAM-mediated screens exhibited a high degree of consistency and validation with >80% effectively...software for analyzing the data. It should be straightforward to apply the new library for use in various...an engineered CRISPR-Cas9 complex. Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena ...

...microscope where the light source is mounted above (epi) the specimen and the excitation light passes through...cause a higher-energy electronic transition in a fluorescent molecule (see figure to the right). The excitation... proteins are excited by 750 nm to 760 nm laser light, enabling dual color imaging studies with blue or...fluorescent proteins (proteins that fluoresce after a light-induced chemical reaction) or photoconvertible Keade... Keade proteins (protein that undergoes light-induced irreversible photoconversion from green fluorescence...

...recommendations that postdoc salaries be raised to $50,000 - higher than the level being raised to now - across a wide...from postdocs and faculty (at time of writing, Brigham And Women's in Boston also just announced that ...before the injunction, are raising salaries, even in light of the injunction. In addition, PIs at some of the...Institute, where salaries had already been raised higher previously and all 6,000+ postdocs at the University...contract - negotiated by their union - securing higher salaries. This decision will carry over into other...information, many assumed that those living in areas with high costs of living would get much better salary increases...

... experience, we found that fluorescent proteins might not always be the best option. Fluorescent proteins...antibodies coupled to nanogold particles, that allow a high-spatial resolution of the protein localization. ...hypothesized that a different tag fused to the hM4Di might improve the transport of this DREADD to the plasma...even when fused to mCherry. But our data does highlight the importance of a careful characterization of... Proteins 101: GFP Fusion Proteins – Making the Right Connection.     Thank you to our guest blogger...

...market because of life-threatening health effects. Eight of these posed greater health risks for women than...cause human suffering and death. Doing research right has the potential to save lives and money. A 2014...innovations It is crucially important to get the research right from the beginning. This is the goal of Gendered...reasons why drugs fail - sex is one of them. Why might the sex of the cell be relevant? Research shows ...disparities in disease severity. Such disparities might suggest underlying sex-based influences on physiologic...curricula. Corporations need to integrate these insights into product design. But eyes have been opened—and...

... mind, when planning an experiment users should weigh the benefits of enhanced biosafety versus a potential...Tet-off transcriptional activation system to drive high levels of expression of certain viral components...manufacturer these changes result in yields up to 25x higher than other available systems. Producing lentivirus...obstacle in obtaining usable lentivirus is producing a high enough concentration, or titer, for your experiment...a double harvest approach at 48 and 72h produces high titer virus for transfer plasmids with small inserts...11589831. Jiang W. et. al.. An optimized method for high-titer lentivirus preparations without ultracentrifugation...

...Although methods to switch on Cas activity using light or drugs have been developed, the field lacked an...anti-CRISPR proteins work? Anti-CRISPRs proteins are highly diverse but most block CRISPR activity one of three...Regulating Acr proteins by inducible promoters, light, or small molecules could achieve rapid, dynamic...allows anti-CRISPR activity to be controlled by blue light (Bubeck et al., 2018). 3. Reduce cytotoxicity of...pathogens possess CRISPR-Cas systems, phage therapy might not be as effective. This is where Acr proteins ...CRISPR Plasmids for Your Research Interested in light-inducible Cas9 activity? Check out these these plasmids...

...overexpression construct is being delivered, that is a higher risk cargo for a researcher to come into contact...essential for cellular function, this would also be a higher risk vector. While it is unlikely that you, or ...other BSL-2 materials, you’ll need to use aerosol-tight caps, compatible with your centrifugation speed,...useful. Some organizations or cities may require a higher percentage of the decontaminate to be used for ...researchers. Note that safety policies will vary slightly from organization to organization, so you’ll need...cut a few corners. For many researchers, viruses might be the most dangerous things they work with, and...

...very powerful for high level recombinant protein expression. These systems enable high expression of very...biology research (eg. polymerases and other enzymes might be purified and used to manipulate DNA), or in medicine... through stable cell lines. Both methods produce high protein yields if transfection is successful. Some...if a desired protein product is toxic to cells at high concentrations. Despite their advantages, mammalian... promoter and induced by lactose, provides the highest level of protein expression. Despite their ease...

... trying to perform high-throughput experiments. If a user wants to carry out a high-throughput genetic...the DSB. CRISPR has allowed scientists to tag and light up endogenous genes of interest to better understand...convenient one tube assembly reaction to generate high-throughput targeting vectors. These vectors can ...fluorescently-tagged proteins at endogenous loci, shedding light on previously unknown protein functions. References.... and Eric M. Jorgensen. “SapTrap, a Toolkit for High-Throughput CRISPR/Cas9 Gene Modification in Caenorhabditis...

...Association for Cancer Research Annual Meeting, you might consider including more background on how your virus...to expertly show off your science.  Prepare the night before: To avoid rushing out the door without your... tack up next to your poster. Arrive early: You might be nervous before your poster session, or it may...walk up to someone’s poster, and then he/she dives right into the background without an introduction. If ... in his/her work, and would enjoy hearing the highlights so that you can appreciate the impact of the ...

... use to introduce yourself and what you do. You might want to include your university and your advisor...new at a conference, one of the first things you might want to know is what their project is. Or maybe ...familiar with. You can tweak your elevator pitch slightly if you know a bit more about the other person....elevator pitch several times at the conference, you might notice some trends. Were there parts of your elevator... up on the board and meet your poster session neighbors. During the session, if someone comes up to my...and your resume and prepare business cards. You might not consider being a student or postdoc a “business...

...Every few months we highlight a subset of the new plasmids and viral preps in the repository through ...CRISPR corner By Jennifer Tsang Here are a few highlights from recent CRISPR plasmids. To find all of the...Plasmids and Resources page.  A new system uses light to deactivate photocleavable gRNAs within seconds... assembly of CRISPR/Cas9 constructs. zCas9i is a high efficiency intron-optimized SpCas9-coding gene. ...

...Every few months, we highlight some of the new plasmids, antibodies, and viral preps in the repository...nucleocapsid RNA chaperone, RT: reverse transcriptase. Right: T1 tomato plants that inherited an edited gene ...rely on synthetic dye molecules, which are much brighter and more photostable than fluorescent proteins...Craggs, T.D., & Twelvetrees A.E. (2024) Kinesin-1 is highly flexible and adopts an open conformation in the...illumination regimes with fast activation kinetics and high responsivity near the resting membrane potential...

.../s12915-019-0682-0. Beck, C., Gong, Y. A high-speed, bright, red fluorescent voltage sensor to detect...the kinetics of voltage sensing to signal output slightly slower than other approaches. Functionally, the...photobleaching, lack of red-shifted indicators, and brightness of fluorescence all have room for improvement...transport, not fluorescence, they are not nearly as bright as the other GEVI systems that use engineered FPs...one component counterpart and is significantly brighter and available with red-shifted fluorophores (find...

...average, AAV.PHP.eB and AAV.PHP.S had 1.5- to 2-fold higher GFP intensity compared to the strains they were...were derived from. They also transduced a higher number of neurons overall (GFP+ cells that also stained...dendrites extend to. The keys to this method are 1) high levels of color diversity so that cells are labeled..., or blue (RGB) fluorescent protein (4a-c). When high doses of this viral cocktail are injected, by chance.... tet-off transactivator (tTA), is injected at a high dose to maximize the number of cells that are labeled...part system, color diversity is maintained at both high (figure 4e) and low doses (figure 4f) of the tTA...

...Kits can also have the annoying tendency to runout right when you need them and to accumulate a bunch of ...precipitated, dried, and then rehydrated. There are some slight modifications depending on the type of DNA you’... not bind silica or glass, but the addition of a high concentration of a chaotropic salt (guanidine hydrochloride...: agarose gel slices Product: DNA fragments You might be wondering, “How is milk going to help me extract...purify DNA ranging in size from 100 bp to 48 kb with high yields and without degradation during the extraction...nucleic acids. Containinants bind cellulose less tightly than nucleic acids, so placing the dipstick into...

...). I’ve assembled a series of screenshots to highlight changes to both the design and functionality of...involves delivering a calcium biosensor using AAV, you might decide you’d really prefer to order ready-to-use...look at filters in a moment. First, I want to highlight the convenient ways our new search tool lets you...you know the ID number for a plasmid, your straightforward search is now even easier with a direct link...