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...so most labs opt to purchase adenoviruses from dedicated adenoviral production labs or companies. Production...something you may have to optimize for your specific application. With mammalian systems, it is standard to do...more cells, while AAV can be titrated following purification using qPCR or digital droplet PCR. Insect cells...to collect your viral supernatant. Baculovirus replicates inside insect cells, and its lifecycle ends in...titer the virus, or go straight into your viral application, like protein production. Titration of baculovirus...Sf9 Time to collection 2–4 days 5–10 days Purification Ultracentrifugation (AAV); centrifugation (lenti...

...RESCUE in therapeutic applications, the authors tested various dRanCas13b truncations that would allow packaging...longer stretches of duplexed RNA. Using an ADAR2DD catalytic mutant, they showed that editing is mediated by...into adeno-associated viral vectors, they tested truncated ADARDDs to see if they could shrink the construct... to deaminate cytidine and used dRanCas13b (catalytically inactive Cas13) to target the cytosine deaminase...construct for viral delivery. They found C-terminal truncations allowed the same or improved editing ability ...

...retroviruses ” (Cullen, 1992) because of their more complicated genomes. Here, we will mostly use the gamma-retroviruses...wild-type viral particle — and of the normal, replicative life cycle — can help you understand these seemingly...of a non-living viral particle, and how we as dedicated researchers can exploit it. This blog was written...Hutcheson for her assistance with lentiviral replication.  Resources and References More resources on...101: Types of Viruses Viral Vectors 101: Viral Applications Resources on addgene.org Addgene's Lentiviral...acid of the Bryan strain of Rous sarcoma virus: purification of the virus and isolation of the nucleic acid...

... Aptamer biosensors are used in a variety of applications, including detection of environmental pollutants... opens aptamers for use in many entirely new applications including aptamer based fluorescent biosensors... render this approach less sensitive in most applications. Learn About Fluorescent Biosensors Light-up..., MG proved to be too toxic for many in vivo applications, but this work proved the concept that RNA could...and biochips. Ultimately, there are just some applications that lend themselves to aptamer-based instead...Kyung-Mi, et al. “Aptamers and Their Biological Applications.” Sensors (Basel). 2012; 12(1): 612–631. PubMed...

...Red arrows indicate a conversion directly catalyzed by the base editor. Black arrows indicate natural cellular...researchers can now select the best tool for their application from among many options.  Conclusion With these...engineered human uracil DNA glycosylase. Nature Communications, 15(1), 4897. https://doi.org/10.1038/s41467...via engineered uracil-DNA glycosylase. Nature Communications, 15(1), 6397. https://doi.org/10.1038/s41467...

... they are based on light which is very easily scattered by tissue. Ultrasound, on the other hand, is a...protein nanostructures called gas vesicles, which scatter sound waves and thereby produce ultrasound contrast...limiting their applicability. They also did not scatter ultrasound nonlinearly, making them difficult to...out to develop next-generation bARGs that could scatter ultrasound nonlinearly and be easily expressed ...promoter pBAD and a low-copy number origin of replication (p15A) worked best under a wide range of inducer...smaller genetic footprint, and nonlinear acoustic scattering. The best candidate for this was the Anabaena...

...a protein’s stability or post-translational modifications, or study how a target protein interacts with...Protein A and Protein G are not suitable for all applications, such as protein isolation from serum. In this...capture antibody interaction and/or the downstream application. Typically, if the target protein will be detected...and an isotype control antibody. Alternative applications Oftentimes, an IP-appropriate antibody simply...identify DNA binding proteins involved in histone modification. Similarly, RNA IP (RIP) can be used to isolate...resources References:  Björck L, Kronvall G (1984) Purification and some properties of streptococcal protein...

...positively charged polycation, polybrene. Interestingly, one study found that alternative polycations such as DEAE-dextran... freshly collected lentivirus for downstream applications. While this ensures that the highest titer virus...lentivirus may want to consider testing a variety of cation and serum sources to ensure the best possible transduction...establishes the best conditions for specific applications, stick with them! Changes in such variables ...efficiency of lentiviral vectors: effect of sera and polycations.” Molecular Biotechnology 53.3 (2013). 308-14...

...EthoVision. Consult the literature to find all the applications of optogenetics relevant to your field. Ultimately...Inhibition of neuronal activation is a little more complicated since basal neuronal activity in your brain region...technology is improving and may one day reach the sophistication of mouse transgenics. Everything that has been... and memory in opioid addiction and science communication/science writing. Derek is also a pianist and...rat lines: tools, techniques, and optogenetic application to dopamine-mediated reinforcement." Neuron 72.5...sensitivity optical neural silencer: development and application to optogenetic control of non-human primate cortex...

...advice on how to dive into new forms of science communication. Click here to listen to the interview with ... from Addgene.      Check out more science education tools like Verse Two and Fill in the Planck at...with us. Tom McFadden is a YouTuber and science educator looking for creative ways to implement the next...next generation science standards and communicate science. Learn more about Tom on his youtube channel ...Community Check out All of Our Podcasts Find Great Educational Content in our Plasmids 101 Series Additional...

...studying genetic modification for their ease of manipulation and their ability to be replicated. Because E...., 2004). Additionally, breakthroughs in gene modification tools and methods such as CRISPR have vastly...Escherichia coli as a Model Organism and Its Application in Biotechnology. In Escherichia coli - Recent...Physiology, Pathogenesis and Biotechnological Applications. InTech. https://doi.org/10.5772/67306 Koornneef... bacteria with superior performance. Nature Communications, 9(1). https://doi.org/10.1038/s41467-018-04899...

...that the oversized AAV genomes are fragmented or truncated when packaged and then following transduction,...and may need to be optimized for each unique application (Trapani et al., 2015).  Expression of smaller...transgene to help prevent the accumulation of such truncated proteins. Examples of use:  Ghosh et al., 2008...design, donor A may be capable of expressing a truncated protein. This can be avoided by excluding a stop...JS (2011) Gene Therapy of mdx Mice With Large Truncated Dystrophins Generated by Recombination Using rAAV6...Improved dual AAV vectors with reduced expression of truncated proteins are safe and effective in the retina ...

...variety of species, cell types, and research applications. Labeling neurons with Brainbow The Lichtman... Brainbow 3.1 and 3.2 (described in the same publication) are unique because by default, they are not ...not fluorescent. Instead, they incorporate a truncated, non-fluorescent form of YFP as the default state...recombination can be detected by immunostaining of the truncated YFP. In Thy1-Brainbow mice, some neurons expressed...level of these XFP is not strong enough for all applications. To resolve this, similar systems have been ...transgene in an adult fly. As a stop cassette is located directly upstream of the fluorescent protein genes...

...variety of small RNA types to choose from and can be catered for your specific experiment. For this blog, we...is an umbrella term for a number of different applications that involve different types of small RNA molecules... by a ribonuclease called Dicer. In a great simplification of the process, Dicer chops off the loop of...sequence is highlighted in green, with red boxes indicating mature siRNA. Hairpin loop is in between the ...sequence is highlighted in green, with red boxes indicating mature miRNA. Hairpin loop is in between the ...for your research arsenal, and there is still a dedicated field of study for improving these tools. Next...

...priorities in most cases include IP protection, publications and funding. If any of these three pillars crashes...budgets, “a large percentage of survey respondents indicated that cost was a significant barrier to ELN adoption...to digital for example): “… 99% of respondents indicated that ease of use would influence their ELN choice...that their research ‘doesn’t fit neatly into one category;”. The study also showed that It is important ...to keep all their data in one place and avoid scattering intellectual property across various software...collaborative functionalities include comments, notifications, sharing files, assigning team members to certain...

...comparison to a control sample. Southern blotting applications range from confirming the results of cloning...interest by recording DNA abundance throughout the amplification process, rather than just at the end as in standard...experiments allowed researchers to identify the location of genes. With this knowledge, researchers were...was added and when. Compiling the identity and location of a given nucleotide, a researcher (or a simple...sequence thousands of reads at the same time. Other applications of sequencing, such as confirming the results...replaced these, these techniques still have useful applications and have informed many current techniques. For...

...and easily navigate to educational resources, tools, collections in our catalog, and the Addgene Help ...search bar. And if you’re in the mood to browse our catalog, you can use the links under the search box to ...scrolling up and down the page.  Figure 3: The Education & Tools menu Further down What happens if you ... mission, links to our deposit, request, and educational resources page, and previews for our three most...

...channelrhodopsin-2 (ChR2), a blue light-activated cation channel, from the archaebacteria Chlamydomonas ...stimulation. GPCR signaling GPCR signaling is a complicated process involving many downstream signaling mechanisms...break your experiments!  First, closely read the publication, including the supplemental data. You’re most... both of which have extensive collections of publications and information on hardware, optogenetics tools...2003). Channelrhodopsin-2, a directly light-gated cation-selective membrane channel. Proceedings of the ...bidirectional dual-color control of neurons. Nature Communications, 12(1), 1–20. https://doi.org/10.1038/s41467...

...site, or when using chemogenetic tools, where modification of cellular activity outside of your experiment.... A. (2023). Various AAV Serotypes and Their Applications in Gene Therapy: An Overview. Cells, 12(5), ...comparative analysis of constitutive promoters located in adeno-associated viral vectors. PLoS ONE, 9(...Approaches and Further Perspectives of AAV Tropism Modification. Pharmaceutics, 13(5), 750. https://doi.org/...Retrograde Serotype Viral Vectors 101: Viral Applications...

...honestly evaluate your own style of work and communication before making this important decision. I’ll ...Prepare by going sleuthing on the lab website, publication records, LinkedIn, ResearchGate or Twitter. Be...the appropriate first authorship position on a publication of her project and made another lab member first...– including failure to acknowledge good work, allocation of meaningless tasks, removal of responsibility...connections such as serving on Boards? Does the publication record show evidence of industry collaboration...skills guide Learn about careers in science communication Read posts on mentoring for scientists Resources...

...piggyBac™ technology can be used for numerous applications, including transgenesis, gene-trap screens, ...of the piggyBac™  transposase/transposon gene modification system and the scarless, Footprint-Free™ removal...technology that may be used for a variety of research applications, including making transgenic cell lines and ...PMCID: PMC2567143. 6. S. Radecke, F. Radecke, T. Cathomen, and K. Schwarz, “Zinc-finger nuclease-induced...oligodeoxynucleotides: wanted and unwanted target locus modifications.,” Mol. Ther., vol. 18, no. 4, pp. 743–53, Apr...Addgene.org Find Vectors with the piggyBac™ Transposon Catch up on Everything CRISPR with Our CRISPR Guide Deposit...

...speak for you (and you for them) Communication Being able to communicate effectively is part of what makes...you can. The colleagues you help will be future advocates and references. Collaborate with another lab,...productive. Take all the opportunities you can to be communicating about science with others. If opportunities ... (be on a speaker committee) Follow industry publications like FierceBiotech to “learn the business” Attend...

...results, modifications, and inventions created by the Recipient Right to file patent applications on modifications... about intellectual property rights, pending publications, and commercial use often come up. An MTA can...trials Acknowledgement in recipient’s future publications relating to the material Limited liability...

...Since the first research applications of GFP were published in the 1990s, biologists have spent a lot...color was observed after excitation at 488 nm, indicating that spontaneous conversion did not occur. Tobacco... Sattarzadeh et al., Sci Rep. 2015 CC BY 4.0 Applications and limitations of GFP photoconversion One concern...2):131-3.  PubMed. Find plasmids from this publication at Addgene. Subach OM, Patterson GH, Ting LM...nmeth.1664. PubMed. Find plasmids from this publication at Addgene.   Addgene’s Fluorescent Protein...

...mostly because of this article: “A systematic identification and analysis of scientists on Twitter.” This...from the United States Standard Occupational Classification System  and Wikipedia. 2. Social and computer...: Portion of Tweets with Scientific Domains, Categorized by Discipline. Biologists, neuroscientists, and... Y., & Sugimoto, C. R. (2017). A systematic identification and analysis of scientists on Twitter. Plos...Scientists 5 Reasons to Use Reddit for Science Communication 10 Ways to Share Your Science Why Scientists...

...working in your chosen field. This cooperative education experience (co-op) is what has brought me, a second... QC/QA team as a DNA Sequencing Intern. The application process When I first saw the comprehensive job...for the intended position. After submitting my application, I was contacted to participate in a brief fifteen...the depositing labs intended. With the use of applications like SnapGene and Geneious, I was able to determine...currently working on projects for the Science Communication function at Addgene. Working a 9-5 As it turned...

...adaptive bacterial immunity and its potential applications in rewriting the genome. Their award acceptance... same Polish sub-dialect making their Skype communications extra fun) who shared first authorship on their...simpler 1 protein - 1 RNA system.  Dr. Charpentier advocated for the crucial role of basic research in the ... area will accelerate the launch of clinical applications. During the panel discussion, Dr. Mello took...for billions of years (…) Bacteria are really sophisticated”.     New to CRISPR/Cas9 Technology? Learn ...

...of hues. While those tools are great for many applications, Verkhusha and his lab at Albert Einstein College...light. As the researchers described in Nature Communications last year, “The capability to control spectral...Verkhusha’s team reported another advance in the application of near-infrared fluorescent proteins, iRFP670...fluorescence is because the light becomes very scattered,” Verkhusha explained. Ultrasound wavelengths,...hand, are larger than the size of a cell, so they scatter much less. As a result, ultrasound enables higher...

... difficult to eradicate. You can suppress the growth of mycoplasma but cannot eradicate it completely....Contamination! Tissue culture media often have indicators included in the media that turns yellow from ...important to prevent contamination rather than eradicating it later. Don’t be scared when mycoplasma infects...the first person to face this kind of problem. Educate others when you can and help them understand the...Cc, Drexler Hg (2002) Comparative Antibiotic Eradication of Mycoplasma Infections from Continuous Cell...

...transfer expert who understands the somewhat complicated Material Transfer Agreement process that kits...most scientific collaborations fall within this category. Unlike a process team, you don’t necessarily ...functioning (or malfunctioning). Clear lines of communication Few people actually like meetings, but run well...are helpful, they are certainly not the only communication options. Addgene, along with an increasing number...desks away or on the other side of the world, communication platforms like Slack make teamwork easier. Trust...excellent leader, and perfectly clear lines of communication, but if teammates don’t trust each other, none...

...scientists work on projects spanning from the identification of plant compounds and insecticidal proteins...bacteria. Learn more about the different CRISPR applications in plants.     Transgenic yeast as source of... yeast capable of desaturating fatty acids. Identification of Insecticidal Proteins Linh Bao Ton (M. Sc...Arturo Rivera Solís, Universidad Autónoma de Yucatán, Mexico Dr. Rodrigo Arturo Rivera Solís, Associate...wide range of research fields and geographical locations. We are excited to see how all of the Addgene-...

...of any scientist’s career. We use writing to communicate our findings and share them with the world. If...simply writing down your data doesn’t guarantee a publication. There is an art to composing manuscripts and...field of microbiology with multiple high-profile publications. My classmates consisted of graduate students...well as being critiqued added an extra level of application not present in the online class.   Four ways ...paper will be generated. After you complete a publication quality figure, produce the figure, its legend...

...job - we strive to also give you high-quality educational resources to help you do better research. CRISPR...design, sorting the plasmids we have available by category. Each plasmid page now has a helpful graphic to... newest page in the Transcriptional Regulation category already has five blue flame plasmids covering ...providing an avenue for temporary/reversible CRISPR modification. In addition to these subpages, our updated ... HypaCas9 Day 10: Transcriptional regulation: Truncated gRNAs for Regulating Gene Expression and CRISPR...

...composed of camels, llamas, and alpacas (and their undomesticated counterparts) - not exactly species that jump...to sdAbs becoming valuable tools for diverse applications. For example, RANbodies can be used as an alternative... excitement around using sdAbs in diagnostic applications. For example, fluorescently and radioactively...heavy-chain only antibodies. (A) Overview of genetic modifications made to generate the nanomouse. (VHH = variable...inspired this versatile family of tools, the applications they have already enabled, and the discoveries...

...tackling these particular tasks, but they tend to be scattered about the internet. To help you keep your virtual...: Find sequences for plasmids assembled from publications and commercially available sources, as well ... as expression patterns, post-translational modifications and polymorphisms Codon optimization tool: To...making it ready to copy and paste into other applications Primer design tools Many of the above-listed... well as primer design for different cloning applications. Primer3 and Primer3Plus: For general primer...

...compounds, links to publication references, and cross-references with cell line catalogs and collections....surveying of cell line collections websites and catalogues Literature searches utilizing Google Scholar ...include accession number, species, and assigned category (cancerous, hybridoma, etc.), as well as sex and...manually going through the pdfs of the 26,000 publications cited. If the information seems incomplete or...

... Specifically, viruses are able to infect and replicate in host cells, but they are susceptible to cells... though not all, of these defenses. For some applications of genome engineering - such as certain forms...to integrate. Additionally, the TA sequence is duplicated near the gene of interest insertion site.   The...1) an SB transposase, the enzyme required for catalysis of transposition; and 2) a transposon containing...containing the gene-expression cassette that can translocate within the genome. The Sleeping Beauty transposase...

... (e.g. 6xHis containing pDEST-HisMBP). After purification the protein can be released from the resin and...types of yeast expression vectors. Consider your applications and experimental design. You can create a new...more traditional approach of an autonomously replicating vector that carries either a wild-type version...a variety of other types of vectors for most applications used to study gene function. From making transgenic...transgenic animals, gene targeting and genome modifications to reporter vectors (e.g. pBV-Luc) and gene ...

...distinct types and subtypes using polythetic classification based on evolutionary similarity (5). Type ...2013 (7, 8). Nine months later, more than 1500 publications describe work to improve the tool’s specificity...various species and describe an assortment of applications. Despite the ubiquity of CRISPR/Cas toolkit,... multiple plasmids used for various platform applications. For more information on CRISPR/Cas, check out...for your research. Do you have a cool idea or application that involves CRISPR/Cas technology? Let us know...

...containing only the sequences required for viral replication and transcription as well as reporter genes. ... membranes. Read the full post for potential applications of R-bodies and to find a video of them in action...Medical School describes how AAVs function and indicates important things to consider if you’re getting...these libraries from the post and the associated publication. Optimizing Donor DNA for Enhanced CRISPR Genome... to your DNA sequence of interest. Potential applications of the system include activation and repression...