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We narrowed to 582 results for: arl

Showing: 61 - 80 of 582 results

  1. Hot Plasmids - October 2020

    Type
    Blog Post
    ...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 ...

  2. Viral Vectors 101: Voltage Indicators

    Type
    Blog Post
    ...: 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...

  3. 15 Hot Plasmids from 2017

    Type
    Blog Post
    ...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...

  4. Hot Plasmids - March 2020 - base editors, GEVI, MoClo, and optogenetics

    Type
    Blog Post
    ...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...

  5. An Introduction to Adenovirus

    Type
    Blog Post
    ...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...

  6. New Viral Vectors - Fall 2024

    Type
    Blog Post
    ...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...

  7. Four Factors that Differentiate the Stem Cell Field

    Type
    Blog Post
    ... 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...

  8. CRISPR Methods for Bacteria: Genome Engineering, CRISPRa, CRISPRi, Base Editing, and More

    Type
    Blog Post
    ...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...

  9. Uncovering Ancestral Bias in CRISPR Research

    Type
    Blog Post
    ...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...

  10. IDP and your PI: A Roadmap for Career Planning and Personal Development

    Type
    Blog Post
    ...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...

  11. Pushing the Limits of DNA Assembly

    Type
    Blog Post
    ...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...

  12. Transferable Skills Guide: Cross-team Communication

    Type
    Blog Post
    ... 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...

  13. CRISPR in the Clinic

    Type
    Blog Post
    ...lymphoma, and some solid tumors are currently in early-phase clinical trials and have shown promising preliminary...et al). CRISPR engineering of CAR-T cells is particularly promising because of its potential for allogenic... polymorphisms (SNPs) in the TTR gene and is molecularly characterized by misfolding of the TTR protein...construct targeting the diseased allele specifically. Early-stage trials showed a high success rate in clearing...the list above! There are many, many more in the earlier stages of the pipeline, ones that have experienced...therapy, since TTR is primarily made in the liver. Similarly, for cardiovascular disease approaches targeting...

  14. Magnetic Control of Proteins: More than a Dream

    Type
    Blog Post
    ...proteins that had shown magnetic responses: EGFP, mScarlet, and AsLOV2. After several rounds of semi-random...semi-random mutagenesis and screening, the EGFP and mScarlet variants were showing no obvious signs of improved...that MagLOV’s “astonishing” magnetoresponse was clearly visible whether expressed in E. coli, in mammalian... tools, they’re optimistic that MagLOV can be similarly adaptable into future magnetogenetic tools.  Embracing...code, raw data, and more. The team shares results early and often, including works-in-progress still in ...passive impact,” he says, noting that his team regularly shares troubleshooting tips and works with other...

  15. AAVs in Retinal Gene Therapy

    Type
    Blog Post
    ... was in its infancy, but the eye was identified early on as an ideal target. It is an easily accessible...dog, was characterized and found to suffer from early and severe visual impairment that faithfully recapitulated...for ocular conditions. However, AdV was abandoned early on due to its considerable immunogenicity and its...candidate therapies for these diseases. Riding on the early successes of retinal gene therapy, an ever-growing... to name a few. CRISPR-based tools are also particularly well suited to be combined with AAV for in vivo...clinical study combining these tools may start as early as next year. Twenty years after falling from grace...

  16. Advancing Biology with Zebrafish: Genetic Tools for Developmental Studies and More

    Type
    Blog Post
    ...zebrafish zygote transforms into an organism with a clearly established body plan including head, eyes, and...lineage tracing defined a high-resolution map of early embryogenesis in time and space, and was also used...mapped using single cell RNA sequencing approaches (Harland, 2018). Several different methods have been developed...popular outside of developmental biology, and are particularly well-suited for neuroscience investigations....Academic Press, 2016. 219-244. PubMed PMID: 27443928. Harland, Richard M. "A new view of embryo development and...Philipp J., et al. "Reconstruction of zebrafish early embryonic development by scanned light sheet microscopy...

  17. 5 Reasons to Use Reddit for Science Communication

    Type
    Blog Post
    ...the best things an early career scientist can do is to get started using LinkedIn early and brush up on ...your science. While Reddit isn’t for everyone, particularly if you’re more interested in selling a product...started watching the questions and comments go up early in the day, and the moderators were on top of any...started watching the questions and preparing answers early in the day. Although I didn’t start answering until...best practices for LinkedIn networking. I was an early adopter of the LinkedIn blog format and it is a ...

  18. Harnessing Bacterial Toxins for Allelic Exchange

    Type
    Blog Post
    ...surrounding genes. However, there are ways to create scarless mutations, ones that don’t leave any undesired...CRISPR-Cas9 technology has also aided in the creation of scarless mutations, both in stand-alone systems (Jiang ...colonies turn subtly blue or magenta. This is particularly valuable when first optimizing conjugation in... Allelic exchange is a great technique for the scarless modification of bacterial genomes. It can accomplish... New Suite of Allelic Exchange Vectors for the Scarless Modification of Proteobacterial Genomes." Applied...Chris R., and Kristala LJ Prather. "The no-SCAR (Scarless Cas9 Assisted Recombineering) system for genome...

  19. Simple CRISPR-based Epigenetic Editing: dCas9-directed DNA Demethylation

    Type
    Blog Post
    ...many of these studies report correlations without clearly testing whether a certain methylation state causes...expression changes (Sapozhnikov and Szyf 2021, Charlton et al. 2020). Most importantly, TET proteins affect...demethylation! With optimization, this method can produce nearly 100% demethylation without the confounds inherent...Read our CRISPR Guide to Epigenetics References Charlton J, Jung EJ, Mattei AL et al. (2020) TETs compete... https://doi.org/10.1038/301089a0. Galonska C, Charlton J, Mattei AL et al (2018). Genome-wide tracking...

  20. Progress Towards a PAM-Free CRISPR

    Type
    Blog Post
    ...cutting most anywhere within the RNA will do. Similarly, if you want to knock out a gene, a frameshift...frameshift mutation introduced anywhere within the early coding region will do the job. What matters for these...you choosing between efficiency and location.   Nearly PAM-less editing   The Kleinstiver lab sought to...and several of the other Cas variants described, nearly all genomic landscapes should be editable with ...Paquet, D., Teo, S. et al. Precise and efficient scarless genome editing in stem cells using CORRECT. Nat...
Showing: 61 - 80 of 582 results