...However, these approaches are often met with the challenges of regulatory bottlenecks and their scope is ...is limited by the lack of well characterized inducible promoters. Far removed from the bio-industry’s vision... various metabolites in Escherichia coli or Saccharomyces cerevisiae with few techniques that are easily...designed scaffold binding sites. With hundreds of characterized aptamer domains available, an unprecedented .... This strategy can be applied to effectively change metabolic flux. The dCas9 protein and orthogonal...6441/ Resources: An open source library of characterized aptamers and supporting publication: http://...

...they change color as they shift from "off" to "on" in the presence of calcium. This color change can be...problem that he considers “one of the greatest challenges of neuroscience.”  Hot plasmids: Twitch sensors...imaging neurons, Griesbeck adds that this color change has a particular advantage for observing events...sensitivity. As reported in Nature Methods, the FRET changes of these Twitch sensor variants were optimized ...trial and error. Transition metals also present challenges in that they are found in cells at much lower...

...through filtration methods because of its ability to change shape and the absence of a rigid cell wall. Here.... For testing, the medium should not have been changed for at least two or three days before sampling ...chemical reactions and this induces a noticeable color change. This method has high sensitivity but low specificity...specific enzymes are used which trigger cellular mechanisms that generate luminescence. This can then be ...are working without gloves or masks. Make sure to change the gloves and masks regularly instead of keeping...water provided to maintain the humidity should be changed regularly as mycoplasma, bacteria, and fungus can...Mycoplasma Contamination in Cell Cultures by Polymerase Chain Reaction. In: Methods in Molecular Biology. Humana...

...questions in the chat box. That’s another benefit to note-taking: you can revisit the Q&A in the chat box. Presenting... plan more with regard to my interests.” Use the chat box and upvote If the thought of walking up to the...asking questions using the polling feature or the chat box. One key thing to remember regardless of the...poster,” he says.Viewing a poster on a screen is challenging, requiring you to zoom in to different parts ...than in-person. You might miss out on the short chats between sessions or waiting for a session to start...

...years. They helped one another navigate career changes and overcome barriers by sharing broader perspectives...Healthcare Businesswomen’s Association (HBA) Boston Chapter and the Massachusetts Association for Women in ...diversity of input and perspectives Provides a good mechanism for accountability that makes mentoring effective... heard from peer groups meeting at both the Fox Chase Cancer Center and Brandeis University. 5 easy steps...choosing a topic and leading the discussion. This is a chance to practice leadership and communication skills...better with process, so don’t just get together and chat. Have an agenda and get everyone ready for the discussion...

... simultaneously   Researchers in the labs of Michael Lin and Ryohei Yasuda have teamed up to develop...maturation. Three new RFPs were isolated and characterized: mScarlet (fluorescent lifetime of 3.9 ns, quantum...member of this class that works via a different mechanism - the PhoCl protein is irreversibly cleaved when... also provided a detailed protocol at Protocol Exchange. Plasmid maps can be found at Addgene and in Figure...Gram-positive bacterium, many of its genes are poorly characterized. The Gross lab hopes these libraries will be...Transcription factor fusions elucidate iPSC reprogramming mechanism The Ptashne lab was the first to demonstrate ...used this now classic technique to elucidate the mechanism by which SOX2 acts to reprogram human fibroblasts...

...Creating epitope-tagged clonal cell lines remains a challenge as it relies on inefficient homology-directed ...representing a protein super family). To overcome these challenges, we set out to develop a hassle-free, highly ...Technologies, USA). These synthetic RNAs can also be purchased from various other suppliers with a typical turnaround...straightforward as the RNA molecules can be directly purchased and there is no need to construct plasmid vectors...; this was possible because reagents could be purchased directly from commercial suppliers without investing...template) and use homology arms of 60 -70 bp . Purchase gene-specific crRNAs and matched ssODNs from commercial...by single-cell deposition in 96-well plates: Characterize clonal lines by ICC and PCR genotyping (Sanger...

...Many thanks to our guest blogger, Doug Richardson! Doug Richardson is the Director of the Harvard Center...This post was contributed by Doug Richardson, Director of the Harvard Center for Biological Imaging and...microscope designs may further exploit the unique characteristics of fluorophores to block out of focus fluorescence...blurry out-of-focus light. The most difficult characteristic to capture in these types of samples is their...model embryos >15 μm Thsee are some of the most challenging samples to image as they require both optical... 2, 932-940 (2005). PubMed PMID: 16299478. 9. Richardson DS, Lichtman JW. Clarifying Tissue Clearing. ...

...lack of caveolin 1 in breast cancer also induces changes of stromal cells in the microenvironment. This ...from cancer cells to the microenvironment. This mechanism happens for both short and long distances. When...transferred to other cells, inducing epigenetic changes in targeted cells. In this case, exosomes may transport...extracellular matrix and fibroblasts. With each cell mechanism discovered in microenvironments, we are one step...References Allinen, Minna, et al. "Molecular characterization of the tumor microenvironment in breast cancer...Intercellular nanovesicle‐mediated microRNA transfer: A mechanism of environmental modulation of hepatocellular ...

...and RNA expression - but what if you just want to change protein levels? Until recently, no such tool existed...proliferation. These cells also did not contain changes in expression of TRIM21-associated genes, indicating...the first author of this article, Dr. Douglas Richardson, recently contributed a relevant post to the ...Addgene blog. Find the SRpHi Biosensors here. Richardson et al. Nat Communications. 2017. PubMed PMID:...transcription factors in series and independently changes inducer levels. That is, if transcription factor...expression of a reporter protein, it is possible to change the amount of noise in reporter gene expression...

...•T-to-G•C base changes (Figure 1), but it can be challenging to make these base changes, especially in...hemoglobin. Previous studies have shown that one base change in the promoter region of gamma hemoglobin (HGB1...effects. ABEs have been described to introduce base changes in cellular DNA and RNA dependent or independent...authors tested ABE8.17-m and found that one base change mutation (ABE8.17-m+V106W) (Gaudelli et al., 2020...

...in comparison to protein-coding genes (Rinn and Chang 2012). Scientists have determined the function ... Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E. A Programmable Dual-RNA–Guided DNA Endonuclease...CRISPR-Based Approaches for the High-Throughput Characterization of Long Non-Coding RNAs. Non-Coding RNA. 2021.... doi:10.3390/ncrna7040079 Horlbeck MA, Liu SJ, Chang HY, Lim DA, Weissman JS. Fitness effects of CRISPR...(5):573. doi:10.1038/s41587-020-0428-0 Rinn JL, Chang HY. Genome Regulation by Long Noncoding RNAs. Annual...in Plants with the CRISPR-Cas Technology is a Challenge yet Worth Accepting. Front Plant Sci. 2015;6. ...

... heteroduplex is cradled in the positively charged channel between the two lobes. For CRISPR to be programmable...light blue; bridge helix, purple.   Apo Cas9 changes conformation frequently (Osuka et al., 2018), but...structure, reflecting the less stringent recognition mechanism that can tolerate mismatches in this region. The...in the duplex that may not fit properly in this channel, depending on their precise location. The end of... BioRender.com.   The exact conformational changes that make Cas9 such a valuable laboratory tool ...Doudna, J. A. (2017). CRISPR-Cas9 Structures and Mechanisms. Annual Review of Biophysics, 46, 505–529. https...

...cancerous cells Luckily, there are several safety mechanisms in place to protect us from rogue cells when ...programmed cell death, or apoptosis, is one such mechanism. When a cell or its genetic material is damaged...daily, yet obstacles remain. One of the biggest challenges facing researchers today is the complexity of... of the latest resources? In response to this challenge, Addgene has launched an initiative to curate ... your thoughts in the comments section below. Michael G. Lemieux is a Scientist at Addgene with interests... Michael G. Lemieux...

...institutional oversight is required. Restraint: mechanical restraint or anesthesia Location: tail vein or...you might after getting a vaccine. Restraint: mechanical restraint Location: thigh Gauge: 25–27 Volume...xenograft models. Restraint: back scruffing or mechanical restraint Location: back or hind legs Gauge: ...Restraint Location Gauge Volume Skill Intravenous Mechanical restrainer or anesthesia Tail vein or retro-orbital...cavity 25–27 < 2–3 mL Intermediate Intramuscular Mechanical restrainer Thigh 25–27 < 0.02–0.05 mL Beginner...Beginner Subcutaneous Back scruffing or mechanical restrainer Back or hind legs; between skin and muscle...

...Did you notice the “Filter By…” options changed when we changed from searching for viral preps instead ...best plasmid for your research can be a daunting challenge. With over 80,000 deposited plasmids from around...assembled a series of screenshots to highlight changes to both the design and functionality of searches...of plasmids? The available filtering options now change dynamically based on the type of material and specific...you may find the available categories and filters change as we now only show options relevant to your current...

...Disable the other repair mechanisms Mammalian cells have 2 primary repair mechanisms besides HR - Non-homologous...mammalian cells, but a less frequent DNA repair mechanism in general. On average, it accounts for less than...is a silent mutation, so you don’t accidentally change your protein of interest.   Fig. 2: Stop...potentially shuttling repair down a different pathway (Charpentier et al., Nat Com).     Fig. 3: HDR events... 8(2024), (2017). 10.1038/s41467-017-01836-2 Charpentier, M., Khedher, A. H. Y., Menerot, S., Brion, A...

...again and possibly mediate HDR. This “second chance” mechanism might improve the frequency of desired HDR...al. added to the CRISPR toolbox with their characterization of two Cpf1 orthologs that display cleavage...requirement. Zetsche et al. started from square one to characterize the Cpf1 nucleases. Using Francisella Cpf1 (...elements of CRISPR-Cpf1, Zetsche et al. turned to characterizing its cleavage pattern. Again, they were in for...efficiency. Kim et al. and Kleinstiver et al. characterized genome-wide editing efficiency of two Cpf1 orthologs...

... LA, Garcia KF, Barnes KG, Chak B, Mondini A, Nogueira ML, Isern S, Michael SF, Lorenzana I, Yozwiak NL...reporter is added to the reaction. Cleavage of the quenchable fluorescent RNA by the “activated” Cas13 produces...Csm6 specific reporter (in the same fluorescent channel as the Cas13 reporter), they could significantly...Donghia N, Daringer NM, Freije CA, Myhrvold C, Bhattacharyya RP, Livny J, Regev A, Koonin EV, Hung DT, Sabeti... Koonin EV (2015) Discovery and Functional Characterization of Diverse Class 2 CRISPR-Cas Systems. Molecular...

...research. This includes chat apps like Slack, Microsoft Teams, and Google Hangouts Chat, resources like Github...an electronic environment and hence making it searchable and shareable, but some ELNs provide better support... delve deeper and look specifically at the characteristics and capabilities an ELN needs to facilitate...send documents from RSpace to a selected Slack channel for discussion and review. It’s also possible to...many different ELNs out there can help with searchability, record-keeping, and data sharing, not all of...