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...    Knocking-down barriers to biotech education: Affordable, low-tech CRISPR/Cas9 experiments By Ashley...Results can be analyzed using either "high-tech" or "low-tech" equipment. Reproduced from Collins & Lau... tumor progression in cancer models. Nature Biotechnology, 10.1038/s41587-024-02551-2. https://doi.org...Ashley Waldron CRISPR/Cas9 has revolutionized biotechnology — partly because of its relative simplicity ...

...apply someone else's new and incredibly useful technique simply by following the abbreviated methods section...They also have plans to expand to other protein technologies later. Organizations such as BenchSci and FPbase...fluorescent proteins so you can best apply these technologies to your own research needs. Other companies ...make it much easier to learn new experimental techniques. Electronic lab notebooks also play a role here...larger datasets and use computer based analysis techniques, an important component of reproducibility is...series of blog posts on the defunct genome editing technology NgAgo). In addition, there are organizations ...

...CRISPR and epigenetics As with many TALEN-based technologies, the advent of CRISPR has made targeting much...modification is certainly not the only CRISPR-based technology designed to alter gene expression. Fusing dCas9... genes from promoters and enhancers. Nature Biotechnology 33:510–517 . https://doi.org/10.1038/nbt.3199...regulatory elements in the human genome. Nature Biotechnology 35:561–568 . https://doi.org/10.1038/nbt.3853...programmable TALE-TET1 fusion proteins. Nature Biotechnology 31:1137–1142 . https://doi.org/10.1038/nbt.2726...modifications at endogenous enhancers. Nature Biotechnology 31:1133–1136 . https://doi.org/10.1038/nbt.2701...

...excellent Technical knowledge and creativity Become a “thought leader,” develop a technical “niche,” develop... something (a journal, a science lunch club, a biotech club) Be the head of a group (like a grad student... develop a useful new assay, master a new technique and be able to talk and own these accomplishments....committee) Follow industry publications like FierceBiotech to “learn the business” Attend events with local...

...come up. An MTA can resolve those questions! MTAs protect you and your institution, and almost all universities... and adopted by the Association of University Technology Managers (AUTM). This MTA provides many benefits...rights – and the rights of your institution – are protected while we handle all the paperwork. Record ...Addgene is internationally recognized as being protective of both the providing institution and the receiving...receiving institution. Therefore, many technology transfer officers are familiar with the UBMTA and are...

...engineering company, Genentech, was founded.  This commercialization of gene editing technology led to the development... Diego. The revolution in genetic engineering techniques is a speculation of yesteryear which has been...double-strand breaks (DSBs) in the genome. These technologies rely on the cellular DNA repair machinery, which...genetic engineering and open advertisement of the technology on various non-scientific platforms, CRISPR has...perspective of the current. Although genome editing technologies have brought a renaissance in the field of health...

...use. In order for this technology to survive the diverse demands of the biotech field, let's look at how...survival tactics of prokaryotes to deliver the technological biological blockbuster known as the CRISPR (...various innovative biological and translative technologies. Addgene has empowered researchers to harness... idea or application that involves CRISPR/Cas technology? Let us know.  References Ishino Y, Hideo,...

...eukaryotes! The technology has been used to optimize protein secretion and expression in the biotech workhorse...once you are up to speed on the basics of the technology. In this blog, we will highlight some common ...applications and kits. Keep in mind that this technology is useful for a variety of purposes beyond what...Heterologous Proteins in Yeast Mitochondria.” BMC Biotechnology, vol. 17, no. 1, Springer Science and Business... Enables Rapid Assessment of Emerging CRISPR Technologies. Cold Spring Harbor Laboratory, 27 Oct. 2023...

...in government, evidence to inform policies, a protected place to work, funding for science, and numerous...cannot prosper in a world where our medicines, technologies, defense, and governing policies are developed...Nuclear Science and Engineering at Rensselaer Polytechnic Institute. He also studies energy markets in ...water. Many marchers held signs demanding the protection of our Great Lakes and our planet. And as we ...the truth about our world. Jonathan Aguilar (@AggleDetector on Twitter) is a graduate student at Johns Hopkins...being taught to communicate in a very specific, technical manner, so much so that we often find it difficult...

...photoactivation - and have enabled sophisticated imaging techniques.     Irreversible photoactivation Irreversible...super resolution imaging, where the power of the technique relies heavily on the same fluorophore being imaged... and electron microscopy (CLEM) is an imaging technique that combines electron microscopy (EM) with fluorescence...fluorescence localization data. However, this technique requires harsh fixation conditions to preserve...fluorescent protein induced by blue light." Nature biotechnology 24.4 (2006): 461-465. PubMed PMID:16550175. ... dual color fluorescence nanoscopy." Nature biotechnology 26.9 (2008): 1035-1040. PubMed PMID:18724362...

...Biotechnology is no longer just for those working in academia or industry. At BosLab, we identify ourselves...towards enabling everyone to participate in the biotech revolution. Our lab exists in what appears to be...Because we’re conveniently located next to a major Biotech hub, building up our lab was easy. Most of our ...engineer yeast to glow, produce synthetic scents, detect compounds in the environment, and openly publish...Kaczmarczyk is a co-organizer at BosLab, a community biotech laboratory in Somerville MA. Follow her on twitter...

...tools by making their own fluorescent protein technologies widely available to the research community. ... a key advantage over other inducible enzyme technologies. She hopes that other researchers will be able... HMOs. Current HPLC-MS and fluorophore-based technologies are too time consuming in the first case and... two biosensors that could quickly and reliably detect HMOs. Fatima’s first design is a whole cell biosensor...Fatima specifically proposes using this sensor to detect HMOs found on cellular surfaces. After incubating...

...Silva, Universidad de Ingeniería y Tecnología (UTEC), Peru Julio Ernesto Valdivia Silva is a master of interesting...identifying new drug targets. An early and accurate detection of PZA-resistance is necessary to improve tuberculosis...engineering: glowing bacteria and a fish robot to detect water pollution Dr. Julio Ernesto Valdivia Silva...synthetic biology resources and microfabrication technologies to generate a self renewable sensor, a bacterium...

...experimented with adding different proteins to the CGBE architecture. They conducted a CRISPRi screen to identify...forms a chemical crosslink with DNA abasic sites, protecting them from cellular processes that would cause...target-library analysis, and machine learning. Nature Biotechnology, 39(11), 1414–1425. https://doi.org/10.1038/...optimization and ancestral reconstruction. Nature Biotechnology, 36(9), 843–846. https://doi.org/10.1038/nbt...targeted DNA transversions in human cells. Nature Biotechnology, 39(1), 41–46. https://doi.org/10.1038/s41587...enable C-to-A and C-to-G base changes. Nature Biotechnology, 39(1), 35–40. https://doi.org/10.1038/s41587...

...it’s true that adding histones and chromatin architecture to every diagram portraying some aspect of eukaryotic...when I recently attended the 2018 Chromosome Architecture and Chromosome Organization Keystone Symposium...posters I saw at the 2018 Keystone Chromosome Architecture and Chromosome Organization. These aren’t the...Crabtree GR Cell. 1998 Nov 25. 95(5):625-36. An architectural role for a nuclear noncoding RNA: NEAT1 RNA ...epigenome. If you have been considering some new techniques, the posts below may steer you in the right direction...

...quite impressed by how neuroscientists are using techniques from across different disciplines to better understand...many opportunities to learn about and develop technologies that will have impacts on many different facets...ready to discuss their VR helmets, gait tracking technologies, and brain monitors. Walking around the poster... addition to the researcher using sequencing technologies to study microbiome interactions with Alzheimer's... or maybe your favorite FRET-based biosensing technique can be applied to an important neuronal signalling...

... cellular targets Optimized genetic screening technique using Cas12a for multiplexed gene perturbation...s41467-019-10846-1.   Optimized genetic screening technique using Cas12a for multiplexed gene perturbation... To validate this optimized genetic screening technique, researchers identified a synthetic lethal interaction...future genetic screens.   DeWeirdt PC, et al. Nat Biotechnol. 2021. doi: 10.1038/s41587-020-0600-6. The Penn...tags, are optimized for SDS-PAGE gels and can be detected on Western blots via engineered immunoglobulin...

...developing assays.  Protocol architecture The basic (very basic!) architecture of the indirect approach is...direct approach isn’t giving you enough signal to detect your protein of interest, it may be time to look...occasion, usually after other signal amplification techniques have been ruled out.  Ease of access Signal amplification...course, this is much easier said than done! Each technique, and sometimes each individual assay, will need...

...and success. Customer and technical support Our customer and technical service brands continue to ... We score ~99.9% helpful and the customer and technical service teams put 100% effort into helping our...also lauded by another set of stakeholders - the tech transfer community. In a recent law review article...our image in the world. Addgene is now as much a tech company as it is a reagent company. I used to have...

...K855A) and eSpCas9(1.1), followed by a technique for detecting genome-wide double strand breaks (BLESS...it’s obvious that CRISPR is a ground-breaking technology that’s already had a huge impact on research ...effects on various biological disciplines, the base technology itself is constantly improving. Cas9 variants... "High-fidelity CRISPR–Cas9 nucleases with no detectable genome-wide off-target effects." Nature (2016...

...recombination. What is Cre-lox? The Cre-lox system is a technology that can be used to induce site-specific recombination... lethality. To facilitate the use of Cre-lox technology, transgenic mice have been constructed that express...system is another site-directed recombination technology very conceptually similar to Cre-lox, with flippase...analogous to Cre and loxP, respectively. The FLP-FRT technology can be an effective alternative to Cre-lox, and...have any tips or tricks, or clever cre-dependent techniques?  Let us know in the comments! References 1....

...President's Council of Advisors on Science and Technology [PCAST] (2012). Engage to Excel: Producing One...Additional College Graduates with Degrees in Science, Technology, Engineering and Mathematics, Washington, DC:...DC: US Government Office of Science and Technology. 3. Duina, A. A., Miller, M.E., & Keeney, J.B. Budding...BioBuilder Use Addgene Videos to Learn about Basic Lab Techniques High School Students Explore Careers at Addgene...Molecular Biology Reference Pages Learn Simple Lab Techniques in Our Protocol Videos ...

...on to find out more about this amazing imaging technique and find further tips for using FRET in your experiments...proteins is not optimal, FRET may not occur or be detected even if the two proteins form a complex. To help...a potential source of error in suboptimal or undetectable FRET, well-characterized FRET reference standards...environment which alters the FRET efficiency. Detecting certain cellular changes with an intermolecular...the concentration or activity of small molecules, detect cellular processes and signaling cascades, quantify...

...viral handling safety   Containment and personal protection are key to lab safety and the same principles...department if they are not currently provided. Eye protection or face shields are required whenever there’s...splash or spray risk in BSL-1. In BSL-2 labs, eye protection is required any time you have to work with BSL... cabinet. Many researchers choose to wear eye protection when they are working in a biosafety cabinet,...organization. It’s always okay to add another level of protection! When centrifuging viruses (a necessary step...

...cell stage using one of several techniques. The longest standing technique is the injection of a targeting...before working with mouse models, several common techniques used to create engineered mouse models at embryonic...the pros and cons of different genome editing techniques. Inbred and outbred strains Mouse strains used...Cre is expressed.  Several variations of this technology can be used to customize the effects of this ...

...science and technology park, it enjoys preferential governmental policies for a high-tech development ...development zone so many science and technology companies and start-ups choose to locate there. I made time to go...because they contain “hard stop” mutations. New techniques used by the Lowe lab have allowed them to begin... and Sun Yat-sen University presented two new technical developments. Using the Tet-on system, his lab...

...making them ideal for single-molecule fluorescence techniques. After generating a series of single- and double-tagged...be imaged using one- or two-photon microscopy to detect action potentials and subthreshold events, including...difficult with GEVIs. Not only was ASAP5 able to detect mEPSPs, it also enabled imaging of their propagation...protein and a NanoLuc® (Nluc) Luciferase-fused detector that specifically binds to active (GTP-bound) ...functional heterotrimers with endogenous Gβγ, enabling detection of activation triggered by endogenous GPCR. Example...

...around the globe. Our first story focuses on a technology that has changed virtually all of biology…CRISPR...repository. Other researchers, eager to try this new technology, were quick to request CRISPR plasmids and, in...favorite observations from this data is that CRISPR technology is consistently showing rapid development and...requested through the years, even as more CRISPR technologies become widely used and available. If we look...all of this work helped scientists use CRISPR technology? We think so — and so do our requestors! CRISPR...

... use various techniques to control protein expression for experimental, biotechnological, and medical ...expression systems Plants provide a cheap and low-tech means of mass expression of recombinant proteins...desired protein. Despite being a rather nascent technology, plant cells have been used to express a wide...expression systems however, are expensive and very technically challenging to use. Alyssa D. Cecchetelli is...

...chromoproteins? One reason may be that chromoproteins protect corals from solar irradiation by reflecting visible...gives scientists the ability for instrument-free detection. Unlike fluorescence or luminescence, which require... fluorometers, or luminometers, chromoprotein detection can be done by the naked eye. This makes cloning...more time consuming and expensive methods. While techniques such as  blue-white screening require X-gal, ... (2000) Fluorescent pigments in corals are photoprotective. Nature 408:850–853 . https://doi.org/10.1038...

... standard EGFP plasmid that used to be sold by Clontech and is in a freezer box in just about every lab...environmental changes that the biosensor is designed to detect resulting in false positives [2, 4]. Solutions ...molecular sizing columns and native gels fail to detect EGFP dimerization. Importantly, all of these assays...assays are performed in vitro. Assays to detect FP dimerization in in vivo cellular contexts, such as ... from nonbioluminescent Anthozoa species. Nat Biotechnol, 1999. 17(10): p. 969-73. PubMed PMID: 10504696...

...To achieve this, scientists use a multitude of techniques including co-transfection of two or more plasmids...started with them. Why Use multicistronic vectors? Detecting cells that are expressing your gene, especially...straightforward process. Rather than try to directly detect your gene of interest, scientists have instead ...not have to exclusively be used as a means of detection; they are useful almost anytime you want to express...upstream of an IRES fluorescent reporter for easy detection.5  Just released: Addgene's Plasmids 101 eBook...

...recombinant Erythropoietin. J Biotechnol. 2020 Feb 10;309:44-52. doi: 10.1016/j.jbiotec.2019.12.017. Epub 2019...conformational changes in the antigen or lower detection sensitivity of the assay. Where possible, it can...beads from very high cell density suspension. Biotechnol Bioeng. 2021 Sep;118(9):3499-3510. doi: 10.1002...Roberts CJ. Protein aggregation - Mechanisms, detection, and control. Int J Pharm. 2018 Oct 25;550(1-2...

...ELISAs. Immunofluorescence. What do all of these techniques have in common? They all typically require secondary.... Could there be an alternative method for immunodetection? Enter the Görlich lab and their anti-mouse...antibodies for all of the above listed molecular techniques, but they can also be expressed in bacteria and...main reasons antibodies are great indirect immunodetection tools is their strong signal strength. This...Although antibodies have been the titans of immunodetection, nanobodies may be better suited for the job...

...between neighboring fluorophores, FRET can be used to detect the inter- and intramolecular interactions that... fluorescence complementation is a method for detecting protein-protein interactions that uses split fluorescent...FP results in a fluorescent signal that can be detected by fluorescence microscopy. Figure 1: How...Medical Center, where he used molecular biology techniques to study inflammation and neurodegeneration. ...BiFC): A 5-year update and future perspectives. Biotech 53. https://doi.org/10.2144/000113943 Additional...

... last 5 years to incorporate more science and technology into the school curriculum. Our Parent Teacher...parents who can answer some of the surprisingly technical questions elementary students can ask. It doesn...and the internet before it, genome engineering technology is moving along much faster than it can be officially...sugarcoat the potential dangers of people using the technology in an unethical manner, but I also tried not ... timely.” Genome engineering, and many other technologies and fields in biology, are moving so quickly...

...Cre-dependent GFP expression was detected with direct fluorescence. GFP was not detected in the absence of Cre. ... are determined using a chromogenic endotoxin detection assay based on the amebocyte lysate method. Purity.... mCherry expression alone was detected. pAAV-Ef1a-mCherry-IRES-Cre was a gift from Karl Deisseroth (Addgene...for mycoplasma contamination using mycoplasma detection kits. Cell line is maintained for ~20 passages...

...specific protein out of a complex mix. Using this technique, users can look for the presence or absence of...acids. Read on to learn more about this versatile technique.  Immunoprecipitation Overview An immunoprecipitation...species as the antibody that will be used for detection in a subsequent Western blot. When choosing beads...application. Typically, if the target protein will be detected in a Western blot then it can be eluted by boiling...epigenetics routinely use the chromatin IP (ChIP) technique to identify DNA binding proteins involved in histone...

...Healthcare consultants advise pharmaceutical companies, biotech start-ups, health insurance companies, and governments...strategy tend to be mostly engaged by pharma and biotech clients. In this realm, there are a few different...organizations, investment and equity research firms, biotech start-ups, venture capital firms, and more. Essentially...science, with a newfound appreciation for how technologies get developed and commercialized after they ...healthcare industry publications like FiercePharma, FierceBiotech, and BioCentury can be an engaging way to learn...

...RNA production can be detected by using a MS2 coat protein that specifically detects a specific RNA stem...instance, pleckstrin homology domains can be used to detect specific phosphoinositides (Varnai and Balla, 2008...Cas9 is a protein, a fusion with GFP allows for detection of genomic locations (Chen et al., 2013). GFP...fluorescent proteins (Chernov et al., 2017) may improve detection. Since longer wavelengths have better tissue penetration...