...real time. Two-photon microscopy was the next revolution in cell imaging, giving researchers the ability...(intravital imaging) studies it’s enabled are revealing the cellular behaviors that mediate adaptive and...proteins (protein that undergoes light-induced irreversible photoconversion from green fluorescence to red... 1. Peters, Nathan C., et al. "In vivo imaging reveals an essential role for neutrophils in leishmaniasis.... Egen, Jackson G., et al. "Intravital imaging reveals limited antigen presentation and T cell effector...

...immune cells harvested from patients and thereby prevent cancer cells from using this receptor to evade ...immunotherapies currently under study. Check out this review for an in-depth look at a variety of up and coming...monoclonal antibody-based therapeutics." Nature Reviews Cancer 15.6 (2015): 361-370. PubMed PMID: 25998715...Lloyd J. Old. "Antibody therapy of cancer." Nature Reviews Cancer 12.4 (2012): 278-287. PubMed PMID: 22437872...Cancer immunotherapy via dendritic cells." Nature Reviews Cancer 12.4 (2012): 265-277. PubMed PMID: 22437871...

...optogenetics. Annual review of neuroscience. 2011;34:389-412. doi: 10.1146/annurev-neuro-061010-113817....effectiveness of your optogenetic manipulations (though a reviewer will probably ask for electrophysiology data too...best suit your specific experiment. Consult these reviews for a discussion of the different variants [1, ...underlying brain disease in animal models. Nature reviews Neuroscience. 2012;13(4):251-66. doi: 10.1038/nrn3171...

...integrate protein tags into the genomes of S. cerevisiae and S. pombe. The protocol is surprisingly simple... in yeast. These tagging vectors are based on previously described pFA6a-link vectors and include a Kan...assessed many of these fluorescent tags in S. cerevisiae, looking at their performance in categories such...a complimentary set of plasmids for use in S. cerevisiae, with the additional benefit of multiple selection...

...Mechanisms. Annual Review of Biophysics, 46, 505–529. https://doi.org/10.1146/annurev-biophys-062215-010822...lobes are inactive in the absence of guide RNA, preventing Cas9 from making off-target cuts.   Pro tip! ...CRISPR–Cas immunity and genome editing enzymes. Nature Reviews Microbiology, 20(11), 641–656. https://doi.org/...High-throughput profiling of off-target DNA cleavage reveals RNA-programmed Cas9 nuclease specificity. Nature...

...brain-visceral organ signaling. The problem  Previously, there were very few tools available for monitoring...   To develop these fibers, the labs looked at previous implementation of optogenetic tools, calcium sensors...stimulation using NeuroStack and the fibers as previously described, then measured electrophysiological...optoelectronics for neural interfaces. Chemical Society Reviews, 48(6), 1826–1852. https://doi.org/10.1039/C8CS00710A...

...real, peer-reviewed study. The second group read the same abstract but with the names reversed, meaning ...looked at how people perceive and evaluate peer-reviewed sources of information if those sources affirm...meaning that the findings of the study were also reversed. The third group read a version of the abstract where...

...of the fluorescent protein dsRed that changes irreversibly its color from green to red when it is oxidized...et al., PLOS One 2013 Oct 7;8(10):e76117. Irreversible peptide-peptide ligation using SpyLigase Building...excitation of neurons. The paper describes five previously unknown channelrhodopsins from different species...designed two vectors for use in conditional and reversible gene silencing in RNAi transgenic mouse models...Jesse Rinehart and Dieter Söll at Yale University previously changed the way researchers can explore important...engineering demonstrate improved efficiency over previous reports. For additional information on using ...

...and exhibited 3-5 fold increased brightness over previous BRET probes, with sufficient signal for in vivo...increased sensitivity and faster imaging compared to previous publications. They also developed calcium, cAMP...described in Suzuki et al. improve the brightness of previous Nano-lanterns by 2-4 fold and permit five-color... its substrate furimazine. In addition to the previously described applications, the enhanced signal of...

... pR, preventing expression of a reporter gene. When light is present, YFI is inactive, preventing repressor...inactive because a bound repressor protein actively prevents transcription. Once an inducer binds the repressor...activation. In this system, the activator rtTA (reverse tetracycline-controlled transactivator) is normally...

...identified have little sequence similarity to previously identified Cas13a-c orthologs, but they do include...system. Since Cas13d is functionally similar to previously discovered Cas13 enzymes - what makes these orthologs...80% for other Cas13a and Cas13b effectors. Like previously characterized Cas13 enzymes, CasRx also displays... manipulate RNA species through exon skipping. Previous CRISPR exon-skipping approaches used two guide...

...reporters in the near future.  SRIRACCHA (stable, but reversible, integrated reporter for assaying CRISPR/Cas-...target region into the N-terminus of mCherry. To prevent alternative start site initiation, the lab removed... this one mutated methionine was not enough to prevent alternative initiation, so they mutated the methionines..., Davidson NO, Madison BB (2017) A stable but reversible integrated surrogate reporter for assaying CRISPR...

...lot of trial and error.” The structural details revealed two exposed RNA loops, which Konermann recognized...of all, the Cas9s can be turned on and off, to reversibly activate or repress genes of interest. They also...in your experiments, Addgene recommends that you review the steps for how to use pooled lentiviral CRISPR... CRISPR-Cas9 complex. Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena C, Hsu PD, Habib...

...treatment regimen and were not preexisting. This was previously debated in the cancer field and was difficult...be amplified using the same sets of forward and reverse primers). The abundance of a particular barcode...replicate; prior to treatment, there are small (previously undetectable) populations of cancer cells that...that combination treatment with multiple drugs reveals even smaller preexisting populations with resistance...

...In previous posts for our Plasmids 101 series, we examined a number of important plasmid elements – promoters...and the region of DNA between the loxP sites is reversed.   Deletion: If the sites face in the same direction... cassette) upstream of a gene of interest will prevent gene expression in the absence of Cre. In the presence...recombination event in a single system. The Saccharomyces cerevisiae FLP-FRT recombination system is another site-...

..., you will need to correct it). It’s easy for reviewers to spot when an application is rushed (whether...that you ask a PI or mentor (your supervisor/ a previous supervisor / someone else in the department) to... times and will have useful advice about what reviewers in your area are looking for. Remember though,...for grant success The two most important things reviewers will be looking for in your research proposal ...

...fluorogen A split fluorescent reporter with rapid and reversible complementation. Tebo AG, Gautier A. Nat Commun...options. While the BiFC interaction is typically irreversible and requires some time for chromophore maturation...maturation, splitFAST displays rapid and reversible complementation. TagBiFC (split HaloTag) provides maximum...important for their interaction. You can read our previous blog post for more on BiFC principles and experimental...

...low cost and quick point-of-care diagnosis kits (reviewed in Tsang and LaManna, 2020). To see even more ...uses loop-mediated isothermal amplification after reverse transcription (RT-LAMP) and a colorimetric readout...completed in just 45 minutes. The team deposited the reverse transcriptase, protease, and Bst polymerase constructs... this toolkit include ones for expressing MMLV reverse transcriptase and Taq polymerase that can be used...

... 1, group D, member 1 EAR1, THRA1, THRAL, ear-1, hRev NR1D2 nuclear receptor subfamily 1, group D, member...

...new managers and I had a “Eureka” moment while reviewing the results of my Myers-Briggs test:  my undergrads...motivation, but he didn’t want to spend hours reviewing papers to piece together the best protocol;  he...associate feedback with end-of-the-year performance review, but feedback should be given throughout the year...References Becoming the boss. Harvard Business Review, January 2007. Three Rules for Managing a Successful...