...and light oxygen voltage (LOV) domains function similarly to cryptochromes with light-induced changes in...Synthetic Neurobiology Group Optogenetics Resources Karl Deisseroth's Lab: Optogenetics Resource Center Boyden...

...temperature on the highest setting. Note: You should see clearly separated layers: Top Phase - Aqueous DNA phase...

...frozen. DMSO helps with denaturation of the DNA, particularly for GC-rich templates. What does each ingredient...

...of phenotypic information was introduced in the early 19th century by Gregor Mendel, who later demonstrated...clones of the original cell. The plasmid DNA is similarly referred to as cloned DNA, and this process of...

...37°C, 5% CO 2 overnight. Although cells should regularly be passaged in DMEM + 10% FBS with penicillin/...

...activity. Read More CRISPR Class 2 C lustered R egularly I nterspaced S hort P alindromic R epeat (CRISPR...

...shown, comparing GFP, wrmScarlet (mScarlet), and the new mScarlet3 and mScarlet-I3 constructs. Scale bar...hours. In order to use the latest mScarlet tags, mScarlet3 and mScarlet-I3, Oliver Hobert’s lab recently...4 protein, mScarlet3 was 50% brighter than the original worm-optimized wrmScarlet. mScarlet-I3, while ...like mScarlet are essential tools for visualizing biological processes in vivo. However, mScarlet’s slow... The team designed coding sequences for mScarlet3 and mScarlet-I3 that were codon-optimized for C. elegans...levels comparable to GFP during early embryogenesis (Figure 3). mScarlet-I3 also offered clearer signals...structural studies of E3 ligase cereblon Optimized mScarlet-based tags for live imaging in C. elegans CRISPR...

...fight the cancer.  Beyond early modifications to standard treatments and early detection, mesothelioma ...the US. Therefore, mesothelioma doesn’t receive nearly as much research funding from the government as...delivered intravenously. New treatment options: Early detection, immunotherapy, and angiogenesis inhibitors...improve the life expectancy of mesothelioma patients: early detection and immunotherapy.  It has long been known... mesothelioma patients is to catch the disease earlier. New techniques are currently being tested in clinical... clinical trials to detect mesothelioma earlier than ever. From breath tests to blood tests, researchers...can come up with new ways to discover the cancer earlier, in particular before it starts to spread beyond...

...studying synaptic connections and physiology is particularly challenging as they are great in number, generally...https://doi.org/10.1016/j.isci.2020.101330 Split-wrmScarlet, a fluorescent proteins system for Caenorhabditis... elegans? By using a split FP approach. Split-wrmScarlet is a new split red FP that is three times brighter...Calico Life Sciences. Like many split FPs, split-wrmScarlet is composed of two FP fragments that are not ... engineered C. elegans cell lines to express wrmScarlet1-10 unattached to any other cellular protein. ...with a ssODN template to introduce the small wrmScarlet11 which is less than 60 nt in length to tag the...distinct localizations. Figure 2: Split-wrmScarlet labeling proteins with different subcellular ...

...: Ace-mScarlet is a fusion between the voltage-sensitive inhibitory rhodopsin Ace2N and mScarlet. Ace2N... change across the membrane of cardiac cells. Similarly, neuronal signaling is carried out by action potentials...fluorescent protein (FP). This class of GEVI was the earliest developed voltage sensor with a genetic component...for ion transport, not fluorescence, they are not nearly as bright as the other GEVI systems that use engineered...Ace2N quenches a proportion of mScarlet's emission via FRET.  Image from Beck et al, 2019 under CC BY 4.0...genetic component, and a small molecule component. Early versions of these sensors contained a FP that localized...quenching of the fluor via FRET. This system is particularly powerful for in vivo imaging especially since...

...characterized: mScarlet (fluorescent lifetime of 3.9 ns, quantum yield of 0.70), mScarlet-I with T74I (...compared to mScarlet, fluorescent lifetime of 3.1 ns, quantum yield of 0.54) and mScarlet-H with M164H...compared to mScarlet, fluorescent lifetime of 1.3 ns, quantum yield of 0.20). All three mScarlet variants...New bright monomeric red fluorescent protein - mScarlet! The Dorus Gadella lab has recently developed ...synthetic red fluorescent protein variant they call mScarlet. Bindels et al used a combination of deliberate...protein fusions. The Gadella lab has deposited all mScarlet constructs with Addgene. Bindels DS, et al. Nat...Methods. 2017. PubMed PMID: 27869816 Listen to our mScarlet Podcast Segment    Photocleavable protein PhoCl...

...protein (mScarlet) with either a seven or eight amino acid linker between them. The Ace-mScarlet voltage...results in higher quenching of mScarlet emission. Figure 2: Ace-mScarlet is a fusion between the voltage-sensitive...inhibitory rhodopsin Ace2N and mScarlet. Ace2N quenches a proportion of mScarlet's emission via FRET. Image ...created two plasmids containing a red-shifted Ace-mScarlet, a fusion of a voltage-sensitive inhibitory rhodopsin...blue-light-sensitive channelrhodopsins. Find the Ace-mScarlet voltage indicator plasmids at Addgene! Beck et...

...genome consists of “early” (E) and “late” genes, and the transition between early and late infection (... AdV transfer plasmid with the E1 and E3 (Early 1 and Early 3) genes deleted. These genes’ functions are... (and therefore between early and late gene transcription) is demarcated by viral genome replication (... (Harrach, 2014). Given that the early genes encode proteins essential to initiating the whole process...only the transgene will be expressed, since the early genes are absent, and the late genes (and their ... knob domain, and hexon), as well as other intracellularly-exposed proteins and downstream cellular events...

...pAAV-Ef1a-DIO mScarlet AAV5 Controls Karl Deisseroth New serotype pAAV_hSyn-PdCO-mScarlet-WPRE AAV5 Optogenetics...serotype pAAV-Ef1a-DIO-ChRmine-mScarlet-WPRE AAV1 Optogenetics Karl Deisseroth New serotype pAAV-hSyn-hChR2...Ofer Yizhar New viral prep pAAV_hSyn-DIO-PdCO-mScarlet-WPRE AAV1, AAV5 Optogenetics Ofer Yizhar New...-hSyn-hChR2(H134R)-EYFP AAV2 Optogenetics Karl Deisseroth New serotype AAV-CamKIIa-jGCaMP8s-WPRE AAV5...

... control process alone takes nearly a year. Addgene depositor Juan Carlos Izpsua Belmonte said it took...able to attend the 10 Years of iPSCs Symposium earlier this year, I jumped at the chance. Here was an ...pluripotent stem cells (iPSCs) capable of forming nearly all cell types in the body from otherwise terminally... prominent role in the scientific endeavor – particularly in fields like stem cell biology where questions...the back burner at scientific conferences. In my nearly 20-year career in science, I have never attended...expensive Dr. Yamanaka explained that it took 50 people nearly 2 years to create the first stem cell line that...

...only 2-3 hours incubation. E. coli and T. citrea scarless editing plasmids Sheng Yang’s lab describes a ...recombineering with CRISPR to create a system for scarless, iterative genome engineering (Jiang et al., 2015...recombinogenic than E. coli, so this system functions more similarly to CRISPR/Cas9 systems adapted for eukaryotes ...expression. The Marraffini lab and Qi lab developed early systems for E. coli; while the Marraffini lab used...Qi et al., 2013). Two important features of this early system are its regulatable nature and its ability...Marraffini lab's CRISPRi/a plasmids In addition to the early work of the Marraffini lab pioneering genome editing...editing using CRISPR in bacteria, they also led early efforts creating tools for CRISPRi and CRISPRa (Bikard...

...remove racial bias from work being done today, is a nearly entirely separate effort from acknowledging its...strongly correlated genetic dependencies that were clearly associated with European or East Asian ancestry...be good targets for therapeutic development, particularly in cancers that are more common in individuals...begin correcting the issue. Working with James McFarland, Jeremie Kalfon, Javad Noorbakhsh, Isabella Boyle...confounding factor when interpreting findings, particularly when researching ancestry groups. And, of course...Adam Brown, John G Doench, David E Root, James McFarland, Rameen Beroukhim, Jesse S Boehm. bioRxiv 2022.11.18.517155...science and society." Science 376, (2022): 250-252. Carlson J, Henn BM, Al-Hindi DR, Ramachandran S. Counter...

...center of her lab (pictured above). DePace holds yearly planning meetings with each trainee to create a...thoughtful planning and honest feedback DePace’s yearly planning meetings revolve around two simple worksheets...tone for the meeting. Another key aspect of the yearly planning meeting is that the trainee leads the ...culture Although they may take some getting used to, yearly planning meetings have tangible benefits for both... to resolve interpersonal conflicts in the lab. Yearly planning meetings may not look the same in every...strategies in the comments section!   References 1. Yearly Planning Meetings: Individualized Development Plans...

...Assembly. Gibson Assembly allows the production of scarless DNA constructs using homologous regions to guide...use of Type IIS restriction enzymes results in scarless assembly. Finally, the NEBuilder® HiFi DNA Assembly...fragment overlap sections, while still creating scarless plasmid inserts.  Want to Learn More about DNA...reset and ligate mismatched overhangs, yielding scarless DNA assembly, and allowing fragments to be taken...used directly in DNA assembly will still yield scarless assemblies. Overview of NEBuilder HiFi DNA assembly...and primer design results in covalently joined, scarless DNA – perfect for transforma­­­tion or as a PCR...

... to realize that there were a lot of words that nearly all Addgene teams used that seemed simple, but ...questions--which were real, not hypothetical--was clearly “no.” When there was room for misinterpretation...list of commonly used (and misused) words that clearly needed an Addgene-specific definition. Words such...setting, where student and postdoc turnover happens regularly and it behooves everyone in the lab to be using..., so we came up with a 3-part strategy: Train: Nearly every new Addgene employee gets “Lexicon Training...Defining words and applying annotation can be particularly useful in academic labs when creating shared...