...discovered the modular format of some transcriptional activators: that the DNA binding domain (DBD) and transcriptional...of whether it interacts with a transcriptional activator (1a) or a repressor (1b). Scientists took advantage... original Y2H systems utilized the yeast Gal4 activator, bacterial LexA DBD and the lacZ reporter gene...transcription-activating function of a eukaryotic regulatory protein. Science. 1986; 231:699-704. PubMed PMID... and Ptashne, M. A eukaryotic transcriptional activator bearing the DNA specificity of a prokaryotic repressor...PubMed PMID: 2547163. 5. Ozawa, T., Kaihara, A., Sato, M., Tachihara, K., Umezawa, Y.l. Split luciferase...

...has a great team of moderators who facilitated scheduling, filtered out inflammatory questions, and even...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...days). Redditors ask serious questions What the moderators did not delete or suppress were the large number...

... concrete happen. You may have more willing collaborators than you realize. Being at a university provides...meetings and take minutes. Get to know your collaborators from other institutions personally on conference...relationship with the perspective of my potential collaborators in mind. Being at a university allows you to...with people at your organization. You can meet collaborators  for lunch, make a point of talking to them ...as messages and keep you up-to-date on your collaborators’ professional developments without directly ...to remind myself to check in regularly with collaborators for example on a holiday or for professional...

...crop species including potato, corn, tomato, wheat, mushroom, and rice (Khatodia et al., 2016). Despite...https://doi.org/10.1128/mmbr.67.1.16-37.2003 Khatodia S, Bhatotia K, Passricha N, Khurana SMP, Tuteja N (2016... al., 2013). Nuclease dead Cas9 (dCas9) based activators (such as dCas9-VP64) or repressors (dCas9-KRAB...

...calcium indicator (Shemetov et al., 2021). In addition to FRET, intensiometric calcium indicators NIR-GECOs...performance, and applications of near-infrared calcium indicators in this recent review (Shcherbakova et al., 2021...encoded near-infrared fluorescent calcium ion indicator. Nat Methods 16, 171-174 (2019). Matlashov, M..... A near-infrared genetically encoded calcium indicator for in vivo imaging. Nat Biotechnol 39, 368-377...Near-infrared and far-red genetically encoded indicators of neuronal activity. J Neurosci Methods 362,...

...expression. Fusing dCas9 to a transcriptional activator like VP64 or VPR activates transcription, whereas... is their persistence and inheritance. CRISPR activators and repressors are thought to be reversible once...cases, epigenetic modifiers may work better than activators/repressors - dCas9-p300 increased transcriptional...enables high-throughput screening for functional regulatory elements in the human genome. Nature Biotechnology...engineering CRISPR toolkit for probing in vivo cis-regulatory interactions in the chicken embryo. BioRxiv: ...Additional Resources on the Addgene Blog Cas9 Activators: A Practical Guide CRISPR Between the Genes: ...

...have begun to use more precise genetic tools: activators, inhibitors, sensors, recombinases, and more....which was recently published in Frontiers in Neuroanatomy.  Here are the three main takeaways from the...achieved in nontransgenic animals by using cis regulatory elements such as promoters and enhancers, multicistronic...multicistronic vectors, and post-translational regulatory elements. Examples of these approaches are given...specificity of the targeted cell population.  Combinatorial strategies can also be used for retrograde tracing...

...cause the least amount of immune response. Any investigator who is using an in vivo animal model should ...identified with AAV2 (5), AAV6 (6) and AAV5 at excitatory connections (3). These facts beg the question...by the use of AAVs? 1) Combined functional and anatomical mapping of circuits In our lab, we use AAV-DREADD...these complexes are so robustly expressed that anatomical assessments can be easily performed too. While...identified this feature of AAVs as a new tool for neuroanatomical tracing (11). With the ability to produce neuron... Floris G., et al. "A fourth generation of neuroanatomical tracing techniques: exploiting the offspring...

...Hendel et al. 2015). Low detection rates mean the investigator can be relatively certain that their samples...off-target mutations are essential data points for investigators looking to definitively link genotype and phenotype...viable options for multiplex sequencing in many laboratories. Nevertheless, unbiased detection isn’t as sensitive...genome-wide unbiased methods. However, most investigators use single cell clones for in vitro CRISPR experiments... Hough is a Biochemistry PhD candidate in the laboratory of Prof Steve Jackson at the Gurdon Institute...Resources on the Addgene Blog Learn to Use Cas9 Activators Perform Genome-Wide Screens with CRISPR Generate...

...CsChrimson::tdTomato, an optogenetic tool (CsChrimson) attached to a fluorescent indicator (tdTomato), as the...intermediate (d), and sparse (e) labeling of the calcium indicator, GCaMP6f, in the Drosophila optic lobe. Image ...dendrites of T5 neurons by placing the calcium indicator GCaMP6f in the effector position, as well as to...

...same page. Ideally, these meetings include all collaborators, from the lab members who are conducting the...interesting results or ask questions to your collaborators. This helps everyone feel included in the project...frequently the entire group will meet. You and your collaborators should then hold each other accountable to these...and circumstances permit, think about having collaborators spend time with each other in person. One possibility...workshops are another great place to meet up with collaborators. Being able to present data from multiple aspects...

...professional-grade design software, like Adobe Illustrator, and I didn’t want to pay for these programs ... Vectr is like a pared down version of Adobe Illustrator. There’s both a web and a desktop version of ...Inkscape Inkscape is the most similar to Adobe Illustrator out of the three options for drawing your own...20,000 icons is designed by professional medical illustrators with input from life scientists. It’s easy to...medical images organized into four main categories: anatomy and the human body, cellular biology, medical specialities...

...pAAV-Syn-FLEX-rc [Archon1-KGC-GFP-ER2] pAAV-FLEX-tdTomato pAAV-GFAP104-mCherry pAAV-mDlx-NLS-mRuby2 CAG-... also be used in conjunction with optogenetic activators and inhibitors that respond to blue light. We...fluorescent-tagged molecular tool.   Find pAAV-FLEX-tdTomato, pAAV-GFAP104-mCherry, pAAV-mDlx-NLS-mRuby2, CAG-NLS-GFP... pAAV-hSyn-DIO-mCherry 59462  AAV2  pAAV-CAG-tdTomato (codon diversified) 37825  AAV8*, AAV9*  pAAV-CAG-GFP...Also coming in our new 20 ul trial size! Voltage indicators Plasmid Serotype Name 119036  AAV1  pAAV-hsyn-flex-Voltron-ST...

..., the researchers did not have access to a P3 laboratory (BSL-3), a high-security facility needed to perform...  Figure 2: When a researcher needs a collaborator for a specific task, Crowdfight will: 1) Find...researchers, such as postdocs and principal investigators. They also come from many different scientific...website. Once curated by our in-house team of collaborators, the requests were forwarded to our volunteer...Biomédica de Salamanca (IBSAL) and one of the coordinators at Crowdfight. She is currently based in Spain...

...effect of mutations of these regulatory elements on gene expression. The regulatory element (wild-type or mutant...gene, such as luciferase, usually serves as an indicator of transcription within cells, where detection... most often used to investigate the effect of regulatory elements, such as promoters, enhancers and untranslated...can then be used to quantify the effect of the regulatory element. Depending on the specific luciferase...

...her then was this: Can you engineer CRISPR-Cas9 activators that work well enough on any gene that they could...which they call CRISPR/Cas9 Synergistic Activation Mediator or SAM. SAM is an engineered protein complex for...information from the structure to guide and improve activator design,” she said. “Before it was a little like...Konermann recognized as an ideal spot to fuse the activators instead of at the protein’s C terminus. With ...repressing or inducing individual transcripts. The activator (CRISPRa) sgRNA library uses the sunCas9 system...

...human pluripotent stem cells: a transcription activator-like effector nuclease (TALEN) kit consisting ...Cell Stem Cell paper, for example, we generated hepatocytes to examine the effect of a gene on cholesterol...Addgene: Tell me about the history. How had investigators done this kind of work before? Musunuru: In ...Unfortunately, it is time consuming and difficult for investigators without a lot of expertise to create zinc finger...and puts it in the hands of pretty much all investigators. Our kit makes it extremely simple to design...

...generation of human hepatocyte-like cells from induced pluripotent stem cells. Hepatology, 2010. 51(1): p....of these cell types [13]:  Germ cells [14] Hepatocytes [15] Pancreatic β-cells [16] Intestinal tissue... intermediate pluripotent state is not always obligatory in cell conversion processes [35]. Rather than...transdifferentiation. Multiple cell types such as fibroblasts, hepatocytes, and pancreatic exocrine cells have been successfully..., A.E., et al., Directed differentiation of hematopoietic precursors and functional osteoclasts from human...

...Schmidt, a Marie Curie Fellow at The Sainsbury Laboratory. Are you a science student or early career researcher...in education advocacy non-profits like Phipps Conservatory or the Brilliant Club and share your knowledge... photo competition. Or nominate yourself as an educator for a short, animated TED-Ed lesson. 6) Network... pros Where do you meet like-minded science communicators? Attend the annual flagship meetings of AAAS...bananas as a Marie Curie Fellow at The Sainsbury Laboratory. She is passionate about science communications...

.... The first was a new class of programmable riboregulators, called toehold switches, which allow for the...allowed gene circuits to be deployed out of the laboratory in a cell-free and biosafe format that was stable...circuit-based sensors to be used outside of the laboratory. The detection threshold for these RNA sequences...the technology through product development, the regulatory process, and manufacturing so that we can get...., et al. "Toehold switches: de-novo-designed regulators of gene expression." Cell 159.4 (2014): 925-939...