... 37°C heat block, LED blue light illuminator or UV light illuminator. Approach: Highly similar to the ...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...

...transcriptional repressor), and dCas9-VP64 (transcriptional activator)-containing plasmids. Each destination vector ...one. This approach is especially useful for combinatorial analyses. For example, if you were to develop...A versatile modular vector system for rapid combinatorial mammalian genetics.” Journal of Clinical Investigation...Cress, Brady F., et al. “CRISPathBrick: Modular Combinatorial Assembly of Type II-A CRISPR Arrays for dCas9...

...pAAV-Ef1a-fDIO-ChrimsonR-tdTomato AAV9 Controls Jensen New serotype pAAV-CAG-2xNLS-tdTomato-WPRE-hGHpolyA AAV9...multiple serotypes pAAV-CAG-FLEX-rc [Jaws-KGC-tdTomato-ER2] AAV5, AAV8 Optogenetics Boyden New serotypes...

...ways scientists have adapted them for use in the laboratory. The discovery of GFP The story of how fluorescent...sequence. Soon after that, in 1994, Prasher’s collaborator Martin Chalfie expressed GFP in exogenous organisms...studying it grew. In particular, studies in the laboratory of UCSD biochemist Roger Tsien resulted in much...

...research as an NIH-funded Principle Investigator in the Kotton Laboratory and is the founding Director of ...Medicine and in the Department of Pathology and Laboratory Medicine. He attends in the Medical Intensive...

...transduction of Cas9-expressing (Rosa26-Cas9) hematopoietic stem cells with a lentiviral vector containing...LaFleur is a Postdoctoral Fellow in Arlene Sharpe’s laboratory at Harvard Medical School and is interested in...G., & Sharpe, A. H. (2024). X-CHIME enables combinatorial, inducible, lineage-specific and sequential ...

...Vampire bacteria Vampirovibrio chlorellavorus is a predatory bacteria that attaches to the cell wall of green...forcing it out of hiding to become exposed to predators. Hungry birds spots the “caterpillar” and then... "Back from the dead; the curious tale of the predatory cyanobacterium Vampirovibrio chlorellavorus." ...

... interpret signals from these reporters. Lack anatomical context/spatial localization: In vivo imaging...years to clinically treat enteric infection and inflammatory bowel conditions. In these experiments, arg1...abdomen. arg1 signal, however, can be overlaid onto anatomical ultrasound imaging to show where the bacteria...

...department.” Caitlin does this by acting as a facilitator between her team, the sales department, and operations...business.” Caitlin’s advice for budding science communicators: “My recommendation would be to take any opportunity...education tools? Check out this Addgene podcast with educator and science rap mastermind Tom McFadden Get tips...

...scientists. Over the course of the class, the moderator discussed individual sections of a paper, the ...quizzes and discussion on that week’s topic by the moderators. The assignments, which consisted of reading ...course was breaking up into focus groups with a facilitator. In the focus groups, we were required to critique...

...passion, and dedication to sharing epitomized by the creators of these plasmids with the first annual Blue Flame...has “literally enabled sharing with 100's of investigators worldwide. Now everything from every paper is...Flame awardee Sophien Kamoun of the Sainsbury Laboratory in Norwich, UK: “We’re very pleased to distribute...

...plasmid. Parts shown in grey are optional cis-regulatory elements.   Key elements of an AAV plasmid...plasmid harbors non-coding sequences. Other cis-regulatory elements Next let’s talk about some of the elements...Woodchuck Hepatitis Virus (WHP) Posttranscriptional Regulatory Element (WPRE) forms a tertiary structure when...

...Willoughby   Using RUBY as a selection marker The creators of RUBY highlight its use in crop plants under... is difficult for new members of the lab or collaborators unfamiliar with these mutant plants. Using RUBY...promoter with self-cleaving 2A site linkages and a terminator. Image from He, Y., Zhang, T., Sun, H. et al....

...Neurobiology has been working on genetically encoded indicators of calcium and other small molecules since the...imaging for beginners Griesbeck says his calcium indicators can be used in mouse, Drosophila, C. elegans,...his team is now applying it to improve upon an indicator they developed and published in 2010 for visualizing...

...to create gene deletions of important unknown regulatory regions in Pseudomonas and Klebsiella spp. The...Ibadan, Nigeriaunder supervision of Dr. Akindele Oluwatosin Adeyi, University of Ibadan, Nigeria Gloria Otito...Candidate working under the supervision of Akindele Oluwatosin Adeyi at the University of Ibadan. She works ...

...15-30 minute break. The method is named for the tomato or “Pomodoro” kitchen timer that the technique’...student.   The Pomodoro timer is literally a tomato! Image from Gerlos.   Tools for checking grammar...with others which makes it easier to work with collaborators. Zotero is a desktop based program that’s compatible...

...culture and include a counter, lab timer, dilution calculator, and a couple of other nice features. This app...sequence for your enzyme of interest. Lastly, the Tm calculator helps you set up an optimal PCR depending on ...apps: Findings Lab Notebook Cell Culture Split Calculator by DRDP Restriction Enzyme Tool by Promega Material...

...histidine kinase that phosphorylates a response regulator (RR). TCS-induced responses are varied; one outcome...conditions, the chromophore activates the response regulator via phosphorylation, promoting transcription of...spread across three plasmids, with the response regulator encoded in the genome. To make the system less...

...identified other advantages, including Cas13d-specific regulatory proteins and high targeting efficiency, both ...WYL domain-containing proteins may be broader regulators of Cas13d activity. This property makes WYL proteins...REPAIR Get tips for using Cas9 Transcriptional Activators Learn about Cas13a (C2c2) Characterization Resources...

...sponsored the capital construction of our new BL2+ laboratory and the infrastructure capabilities to produce...our generous, sharing depositors. Over 3,000 laboratories are sharing plasmids (with 16 labs currently...helping thousands of sharing scientists from laboratories all over the world. With this reach and exposure...

...of stakeholders from industrial and commercial operators, government agencies, active researchers, as well...understanding of the societal ramifications of laboratory science, and how it may best be leveraged for...re always delighted to engage with potential collaborators and interested audiences. Please visit our team...

...effectively contribute as part of a team.” As a seasoned educator, Kuldell also knew that the best days in the classroom... or small, receive the BioBuilder textbook, a laboratory kit, and support during the academic year from...state-of-the-art lab embedded in LabCentral’s community of innovators that opened in August 2017.  This flexible teaching...

...in Huntsville explains it, a major goal in his laboratory is to understand the function of the non-coding...effort to track down every last one of those gene regulatory proteins and the places in the genome where they... that, but the system is so flexible. Other investigators can easily adapt these vectors for use with ...

...containing the RNA pol III promoter, gRNA and pol III terminator. These relatively large cassettes (considering...Size is also a factor; the pol lll promoter and terminator need be encoded only once, and each <200 bp tRNA-gRNA...deletions of other regions, such as noncoding RNAs or regulatory elements, to examine their function. Bacteria...

...(Shaner et al, 2008). Obligate dimers (such as dTomato) that promote interactions between the FPs interfere...CytERM-mKOkappa, CytERM-mTurquoise2, and CytERM-dTomato. When using and interpreting the results of the...proteins. Fusions of CytERM with mTurquoise2 and dTomato, which are established monomeric and dimerizing...

...reporting and genetically encoded fluorescent voltage indicators (GEVI) are ideal tools to track bioelectric states...channelrhodopsin used as a sensitive fluorescent voltage indicator with near-complete desensitization in continuous...Use of genetically encoded, light-gated ion translocators to control tumorigenesis." Oncotarget 7.15 (...

... Laboratory management software is not a requisite for a functioning lab, but it is for a scaleable lab...about what steps are most likely to fail. Our laboratory manager would definitely be interested to see...this case, the physical considerations of the laboratory make object composition a smarter choice than...

...Library, deposited by Carrie Bhang, a research investigator in the In Vivo Pharmacology group at Novartis...ClonTracer Since starting her new role as an investigator in the In Vivo Pharmacology group at Novartis...Addgene community! Carrie Bhang is a research investigator in In Vivo Pharmacology at Novartis Oncology...

...the field. 3. Highlight your network and your collaborators It’s a good idea to move around to different...Networking for Scientists Series 4. Become a science communicator More and more fellowship applications are asking...on your grant application to your peers and collaborators can also help you get some critical insight ...

...), Antibodies: a Laboratory Manual, 2nd Edition. 2104. Cold Spring Harbor Laboratory Publications, New...production and subsequent purification.  Academic laboratories and companies have already begun creating plasmids...

...They play important roles in many cellular and regulatory processes, but much of their functionality is...effects can be minimized by disrupting lncRNA regulatory elements that are crucial for transcription and..., on the other hand, recruits transcriptional activators such as VP64, SAM, or SunTag to specific ncRNA...

...with a terminator. A simple TU could be comprised of just three parts: promoter, gene, and terminator.   ...this category also includes promoters, UTRs, terminators, etc. Once selected, individual units can be ...

...multiple cell types of the same lineage. Blood (hematopoietic) stem cells are a good example of multipotent...numbers or replenish dying cells. For example, hematopoietic stem cells found in the bone marrow only differentiate...three germ layers, form embryoid bodies, and form teratomas [11]. iPSCs also possess similar morphologies,...

...air conditioning on) so leave them in a sterile incubator. This also means that any medium or washing buffer...entering the hood and when putting them into the incubator). It also helps to get familiar with the morphology...results. Most cell culture media contain the pH indicator phenol red (pH range 6.0 (yellow) - 8.0 (red))...

...important supporting roles. Enhancers are cis regulatory DNA sequences that, when bound by transcription...CAPTURE (CRISPR Affinity Purification in situ of Regulatory Elements) method. The original CAPTURE method...2020). CRISPR Affinity Purification in situ of Regulatory Elements: The original CAPTURE method The original...

...  Is this department/laboratory collaborative with other departments/laboratories? How long is the average...by meeting with professors - a/k/a Principle Investigators (PIs) - whose labs does research that aligns...

...localization, dynamics, and behavior of a POI. As an investigator, you want a fusion tag to be inert, to not produce...has gone into making FPs monomeric, yet many investigators remain ignorant of FP dimerization. Equally ...Einstein College of Medicine in the Department of Anatomy and Structural Biology. His interests include the...

...expressing eGFP under the human ubiquitin C promoter. Anatomical abbreviations: Gr= Gracile tract, Cu= Cuneate...  The first successful attempt to render fixed anatomical preparations transparent was done by Walter Spalteholtz...protein (GFP) (6).   The Dodt and Frank Bradke laboratories later developed the 3DISCO (3-dimensional imaging...

...genetically encoded calcium indicators (e.g. GCaMP). Others include: pHluorin & pHTomato (pH sensors), HyPer ...fluorescent intensity upon pH changes (e.g. pHluorin, pHTomato). Keep this list handy as you plan your next ...

...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...Resources Cross team communication  External collaborators For further advice on highlighting your transferable...