...installment in our podcast series, we chat with new Addgene Board Member, Michael Koeris. Dr. Koeris did his graduate... capital firm, Flagship Ventures, Koeris helped change Novophage's focus. The company went from developing...pathogens in food products. Along the way, the company changed its name from Novophage to Sample 6. To learn ...

...excited to announce the recipients of this year’s Michael Davidson and Roger Tsien Commemorative Conference...conference of their choice while honoring the late Michael Davidson and Roger Tsien who have played enormous...fluorescent protein tools. This year, we’ve made some changes. In addition to supporting in-person conference...health and disease,” says Aggarwal. This biosensor changes fluorescence based on the release of glutamate ...colleagues Filip Tomaska and Tim Hanson, are characterizing variants of iGluSnFR3 in vivo and have used...be transferred to the roGFP2 cysteine thiols and change its fluorescence properties. The biosensor can ...allow us to better understand how neuron activity changes following opioid withdrawal, providing insight ...

...Susanna Bachle Anion-conducting channelrhodopsins (ChR) are ion channels that can be used to silence the...PubMed PMID: 30166441. MerMAIDs - marine channelrhodopsins for the optogenetic toolbox Article contributed...

...Listen to the photoswitchable protein podcast segment Reversible control of photoswitchable proteins, which...the light- and dark- states of the photoswitchable cPYP, which changes in conformation in response to blue...picking approach to select cells with variant characteristics they wanted, the team identified Archon1, a... Discovery of new protein binder tools for photoswitchable proteins cPYP and AsLOV2 Article contributed... which change in protein conformation in response to light, offers a way to spatially and temporally control...structural knowledge of the proteins. To overcome this challenge, the Uppalapati and Woolley lab developed a phage...bind to only the light- or dark- state of the photoswitchable proteins offers a powerful approach to control...

...Young-Gie Chung, Mi-Yoon Chang, Baek-Soo Han, Sanghyeok Ko, Eungi Yang, Kwang Yul Cha, Robert Lanza, and Kwang-Soo...Plasmids for Stem Cell Research Delivery Method Mechanism Efficiency Safety Pros Cons MMLV-derived Retrovirus...element that transposes via a “cut and paste” mechanism between a vector and chromosomal DNA. PB has a...be removed by re-expressing the transposase. A challenge of using PB to generate iPSCs is that genomic ...the cytoplasm independent of the cell cycle. A challenge of using Sendai is that since it’s replication...clones tested had plasmid integration. Another challenge is that multiple transfection makes it difficult...plasmids due to lower activation of DNA-silencing mechanisms. While minicircles are typically smaller than...

...activity, channelrhodopsin-2 (ChR2), a blue light-activated cation channel, from the archaebacteria Chlamydomonas...light, this channel activates neuronal activity by opening and allowing positively charged ions to rapidly.... Light-gated ion channels directly open and close when exposed to light and change the membrane potential...receptor or pathway.  Light-gated ion channels Light activated ion channels (rhodopsins) either excite or inhibit... range of wavelengths. Excitatory channels allow positively charged ions such as Na+, K+, and Ca++ to ...Tichy et al., 2019), photoswitchable tethered ligands (PTL), and photoswitchable, orthogonal, remotely-tethered...with each light source used.   Channel and activation kinetics Some channels turn on and off very rapidly...

...particularly new. Chapter 2, “The Professional Scientist,” addresses the challenges of succeeding in science...Do-It-Yourself Science” described in Chapter 3, and of course the rapidly changing world of genome engineering ...call bad science.”  The three chapters in this section describe the challenges of trusting science in a “...science’s biggest challenges over the coming years. It is arguably the most subjective chapter of the book,...to. The problems and challenges described in The State of Science have not changed in “COVID times,” but...scientists to change their priorities. For many scientists, COVID-19 will fundamentally change what they ... that may have warranted their own chapters, and certainly chapters that could warrant their own books...

...committee, I became Vice-Chair when the current Vice-Chair stepped down. When the Chair then stepped down, ... your own development as a leader. Chairing a committee I was Chair of the Advocacy Committee at the NPA...member. The NPA runs its committees through Chairs and Vice-Chairs that lead them. As a committee member, ...the other members on the team and keep the Chair and Vice Chair of the committee updated with their progress...became interim Chair automatically as it was an understood expectation of any Vice-Chair. Following an ...there were many changes to the  Board of Directors and the Executive Director changed twice. I had to ...Leadership Institute” (CLI) where each Committee Chair and Vice Chairs were invited to spend a weekend together...

...real-time, posing a huge challenge to studying epigenetic changes over time.  This challenge was difficult to ...the study of heritable changes in gene expression that do not involve any changes to DNA sequence (Epigenetics...modifications can result in phenotypic changes without any change in genotype. Epigenetic modifications... within an organism. Mechanisms of epigenetic modifications The main mechanism of epigenetic modification...DNA is wrapped around histones changes, then gene expression can change as well. The more frequently and...epigenetic changes were statistically significantly associated with reduced life-span, but no changes in the.... Chromatin remodeling can occur by two main mechanisms, either through histone modifications, which ...

...chloride channel SwiChr++ - A channelrhodopsin converted into a chloride-conducting anion channel. It is...the role of ion channels and pumps in developmental biology.  Examples of some ion-channel and ion-pump ...ChR2 (blue-light activated cation channel) light-activated channels to hyperpolarize tumor cells, which...BLINK2 potassium channel - An optimized blue-light induced synthetic potassium channel. Upon activation... can be measured as the imbalance of electrical charge between the inside and the outside of the cell....cell. This imbalance is maintained by ion channels and pumps in the membrane. An emerging interdisciplinary...role of bioelectrical signaling mediated by ion-channels and pumps in developmental biology, regeneration...

...when a B cell changes which heavy chain constant domain it pairs with its variable chain domain. B cells...of an antibody labeled with Fc, Fab, heavy chain, light chain, constant region, and variable region. ...in their heavy chain constant regions result in several unique physical characteristics of these five ...by the heavy-chain constant genes μ and ẟ. These are the first two genes in the heavy-chain constant domain...isotype? An isotype determines several key characteristics of an antibody as well as the role it plays...class of antibody that’s determined by its heavy-chain constant region (see Antibodies 101: Introduction...antibody isotypes that each have a unique heavy-chain constant region: IgM, IgD, IgG, IgE, and IgA.    ...

...annurev-neuro-111020-100834 Challis, R.C., Ravindra Kumar, S., Chan, K.Y., Challis, C., Beadle, K., Jang, ...spiral ganglion neurons Mice, rats No (mechanism unknown) Chan et al., 2017 Asencor et al., 2022 ... Deep Parallel Characterization of AAV Tropism and AAV-Mediated Transcriptional Changes via Single-Cell...10.1016/j.ymthe.2019.02.013 Huang, Q., Chan, K. Y., Tobey, I. G., Chan, Y. A., Poterba, T., Boutros, C. L....or lumbar spine. Image and caption courtesy of Challis et al., 2022. Where do they come from? Systemic... (>1 x 1012 vg total in mice) may increase the chances of inducing an immune response against the capsids... capsid in your strain of interest, check the mechanism of entry if known, and, as always, validate the...

...composed of four peptide chains (two identical heavy chains and two identical light chains) each with a constant... between one heavy chain and one light chain. When assembled, the four chains make a Y-shaped protein ... The chains are covalently linked to one another by disulfide bonds between the two heavy chains and between...comprise the entire light chain and a variable and constant region of the heavy chain. As the name implies,...= Variable domain. L = Light chain. C = Constant domain. H = Heavy chain. Created with biorender.com   ...Fab’ fragments are almost interchangeable, but the extra bit of heavy chain on a Fab’ can act as a convenient...portions of the constant regions of the two heavy chains. In the lab, this is the region that determines...

...building block for further chaining. The MXS-chaining kit contains 5 empty chaining vectors plus a destination...large changes in binding affinity with light stimulation. Indeed oLID showed only a two-fold change in affinity...binding assays, to engineer photoswitchable dimers which show over a 50 fold change in binding affinity with...with MXS-chaining. Analogous to modular assembly methods, such as BioBricks, the MXS-chaining method uses...are also ready for further chaining. Plasmids constructed using MXS-chaining were used in a variety of ...order to determine the relationships between pH changes and cellular events. To overcome the issues with...(BRET) were developed by the Johnson Lab . Characteristics of the pHlash reporter protein include: H+...

.... Congratulations to Paula Videira, Michael McMurray, and Richard Gronostajski who each are being awarded...27393411. Michael McMurray: Rescuing arginine mutations in rare disease with guanidine Michael McMurray...PMID: 9514755. Richard Gronostajski: NFI-related diseases of neural development Richard Gronostajski is...manifestations. PMM2-CDG is the most frequent CDG type characterized by deficient phosphomannomutase (PMM) activity...suffer from immunological defects. These are characterized by a predisposition to severe and recurrent ...surprising. However, the molecular and cellular mechanisms behind the immune defects in CDG patients are...The primary focus of his lab’s research is the mechanisms by which proteins assemble into functional multi-subunit...

...many have mechanisms in place to prevent the burden from getting overwhelming. These mechanisms include ...Cases present specific operational or commercial challenges or questions that clients face imminently. Consulting...on analyzing the situation that presented the challenge at hand and help resolve it through well-informed...this realm, there are a few different kinds of challenges that consultants get called upon to tackle, depending...to collaborate whenever it faces an important challenge. The types of projects mentioned above are just...rewarding parts of the job is being confronted with challenges that are crucial for the success of a drug’s ...reasons why client situations might be especially challenging. For instance, the client might need to get a...

... looking for a flow antibody, the YCharOS (Antibody Characterization through Open Science) initiative ... community produces antibody characterisation data as part of the YCharOS open science ecosystem. It has...initiative has now started producing flow cytometry characterization data for selected antibodies, using the knockout...antibody selectivity. Additionally, it can be a real challenge to troubleshoot RNAi failures. Potential explanations...cell survival or proliferation, an additional challenge is that only partial or ephemeral knockdown will...confirmation of antibody selectivity even more challenging. Correlation with RNA or Proteomic data from ... control containing treated cells. The expected change in expression should then be reflected in antibody...

...Cascade-Cas3 mechanism. Created with BioRender.com.   The reeling mechanism The reeling mechanism is carried... DNA in a number of ways, from small edits that change single base pairs, to chromosomal inversions and...light. One lesser-known Cas is waiting for its chance in the spotlight — Cas3. Cas3 can induce genomic...recruit Cas3, which carries out the brunt of the mechanical work for editing. Class 1 CRISPR systems make...of CRISPR-based immune systems in bacteria and archaea. Class 1 is further broken down into different ... contain Cas6. Regardless of the subtype, the mechanism of Cascade binding and subsequent DNA degradation...this complex, developing Cas3 tools has been challenging. It can be difficult to ensure comparable expression...

...to look at the other channels you’re using first. A different fluorophore channel may be affecting your... be challenging! But we hope this guide can help you get started. Remember, you can always change your...process of correcting spillover from one fluorescent channel to another. When you label your samples with multiple...similar wavelengths (See this handy emission spectra chart to find your antibody's spectra). Your cytometer...population. By compensating between fluorescent channels, you can correct for this spillover and more easily...using a tool like BioLegend’s Fluorophore Guide Chart.  Pro tip! Some antibodies go by different names...look like and how do I fix it? When plotting two channels against each other, you should be able to draw...

...abundant, charged tRNAs but will stall at regions calling for low abundance, poorly charged tRNAs. When...different codons, modify the tRNAs correctly, and charge the tRNAs with the appropriate amino acids. If ...of codons, it only needs to produce a subset of charged tRNAs and therefore may need fewer resources for...encoded by codons with low abundance or poorly charged tRNAs may be produced at a lower rate than proteins...proteins encoded by highly abundant, charged tRNAs. For example Tuller et al. found that translation efficiency... by a mixture of codons with highly and poorly charged tRNAs, different regions of the protein may be ... this may give the swiftly translated regions a chance to fold properly. For example Pechmann and Frydman...