...optimized for dual-AAV systems. These variants use different RT enzymes and performed large edits with similar...types. DIRECTED is compatible with fusogens from different viral families and can be used to target multiple... to reduce negative side effects. This platform offers a promising approach for experimental therapies... enzymes, TurboID and APEX2, are localized to different cellular compartments. They showed APEX2 can use...uses sequential labeling by TurboID and APEX2 at different cellular locations. C) Enrichment of dual-labeled...stably or inducibly express tagged iPAK4 from different promoters, specific time points appear as fluorescent...bands of color with HaloTag-iPAK4 by applying different HaloTag dyes and showed they could record transcriptional...

...many countries, import and export regulations, differing customs processes, and the distances the packages...distributes to per year, 2005-2023. We identified and offered a number of solutions that increased accessibility...each courier we contracted with so we can always offer the fastest shipping. However, there were issues...specific to that country. ‌For example, when we began offering temperature-sensitive materials, we noticed a ... backbones, lentiviral packaging systems, and luciferase expression. On individual material pages, we ...educational resources for each type of resource we offer.  In 2013, we launched the Addgene blog for topics...particular topic into beautifully designed and easy-to-reference eBooks. Figure 3: Global readership of the Addgene...

...acquired drug resistance are referred to as methods of ‘horizontal gene transfer’. Resistance can also arise...you! Many thanks to our Guest Blogger, Jennifer Rivers! Jennifer Rivers ScM, MLS(ASCP)CM is a research ...This post was contributed by guest blogger, Jennifer Rivers, a research microbiologist specializing in...multi-drug resistance are resistant to three or more different classes, or types, of drugs used to treat them...either through random mutation or horizontal gene transfer, means to defend themselves from these naturally...resistance can arise, and has arisen, for many different reasons. Exposure to antibacterial drugs provides...science communication, and public health.     References 1. O'Neill, J. "Tackling drug-resistant infections...

...patients suffer any less acutely.  The Rare Genomics Institute works to help those who suffer from rare...in diabetes and lymphoma research or the many different ways you can improve your heart health. These ...disease research and provide hope to patients suffering from these devastating afflictions. Elia Di Schiavi...abdomen and affects roughly 20% of mesothelioma sufferers. This disease is less widely understood and often...for peritoneal mesothelioma and thereby relieve suffering from this little-known disease. You can find more...Nabavi and will continue to work to help those suffering from rare diseases. These underrepresented individuals...rare disease research funding opportunities. References 1. Jones, C., et al. "PP09. 1–2352: Systematic...

...through the proliferation of specific, and not all, Langerhans cells, to create foci of proliferating cells....construct containing fluorescent proteins (XFP) of different colors to label an organ or entire organism. Because...more refined versions have been developed, which differ in the number, stability, and brightness of the...localization sequence, allowing it to localize to a different area of the cell than CFP and YFP. The most recent...could recombine to activate the expression of a different fluorescent protein. Therefore, each cell could...become a powerful tool for understanding cell proliferation. Inducible XFP expression in the Ubow mouse ...can be combined with new imaging techniques.  References Cai, Dawen, et al. "Improved tools for the Brainbow...

...through a tiny pore by measuring differences in ion flow caused by the different electrochemical properties ...fragmented into smaller pieces, run on a gel, and transferred to a membrane for analysis through hybridization...standard PCR. This allows the researcher to compare different DNA sequences to see which is more abundant in...content by comparing the fragments created by different restriction enzyme digests. Incubating a DNA sample...restriction mapping (Figure 1). Though some labs may prefer to send DNA directly for sequencing (more on that...incorporation of a ddNTP. Researchers perform four different reactions, one for each radiolabelled ddNTP, and...machines (9). Today, similar technologies, sometimes referred to as sequence DNA as it synthesized by electronically...

...understand genetic cytoplasmic factors that could be transferred between cells. At the time, these extranuclear...episome seemed appropriate, especially since the Fertility, or F-factor discovered by Ester Lederberg in ...-factors. Like F-factors, R-factors could be transferred between bacteria via cell-to-cell contact; however... the same. Over the next few years genes from different bacterial (and eventually mammalian) species were...ubiquitous tool that can be diversely applied to many different experiments. Addgene was founded in order to store... keeping with our goal of creating a one-stop reference guide for plasmids we've updated and added new...: 24043817. PubMed Central PMCID: PMC3785787. References: Lederberg, Joshua. "Cell Genetics and Hereditary...

...specific problem. Every species is so different – an awake monkey is different from a cell in a dish. I always...of life. Our most recent paper in Nature Methods offers the first true two-color pair. We'll be sending...want to activate two sets of neurons and see how different parts of the brain or the cells within them work...set of neurons, maybe we could treat patients suffering from Parkinson's, chronic pain, deafness, or other...Boyden: I think because neuroscience has so many different goals: to understand how a mouse changes emotions...find ways to make these tools more selective for different ions. Right now, we can modulate electricity. ... click the link in the Addgene Alerts box. References: Independent Optical Excitation of Distinct Neural...

...cells (Ma et al., 2015). To do so, they fused a different fluorescent protein to various Cas9 variants, ...either MS2, PP7, or boxB) that each recruits a different fluorescent protein (BFP, GFP, or RFP) to the ...green, or red readout. But gRNAs containing two different hairpins will produce a secondary color (yellow...allows for simultaneous visualization of sites on different chromosomes in living cells, the system requires...they chose an intergenic DNA region (IDR3) as a reference loci to measure inter-locus distances of loci ...sgRNA-Sirius-4X(MS2-PP7) showing recruitment of different fluorescent proteins. (B) CRISPR-Sirius used to...try. Find CRISPR-Sirius plasmids at Addgene. References Ma, Hanhui et al. “Multicolor CRISPR labeling...

...research program (think a subtle difference in mouse chow, a different water filtration system, environmental...outline and refer back to the original paper that everyone in your lab has been referencing for the last...original researchers actually did. Admittedly, the differences are often very subtle, and it can take a lot ...and it was just that they were doing something differently. Communication comes in many forms—from transparency...other publishers and organizations have various different versions of checklists and guidelines to help ...rewarding good scientific citizenship in all its different forms as part of graduation, hiring, funding, ...

...not exert force by dynamically rearranging into different polymerization states. Instead, R body monomers...natural context or 3) be modified to extend at different pH levels. Domesticating R bodies We have shown...that R bodies can function to break membranes of different organisms. And finally, we demonstrated that R... can be “tuned” with mutagenesis to extend at different pH values. Using a screen that takes advantage... fact that coiled R-bodies sediment in certain buffers but remain soluble when extended (see figure to...region produced R-bodies that could extend at different pHs. Plasmids expressing expressing both wild ...School. Follow her on Twitter @jessicapolka.     References 1. Kanabrocki, J. A., Quackenbush, R. L. & Pond...

...deposits to Addgene.  Insomuch as different journals represent different research areas, target readers,... would also be great to know how much of that difference in citations is caused by depositing a plasmid...wouldn’t be causal. To control for a number of differences between those papers with Addgene plasmids and..., so our estimates now take into account any differences in citations due to the journal of publication... the same journals! So, is all this remaining difference caused by depositing the related plasmids with...it is still suggestive that we see such a big difference in citations after controlling for journal. And...Visit his website at: http://www.neil-t.com/ .   References 1. Cong, Le, et al. "Multiplex genome engineering...

... labs (see table below) have different PAM requirements. These different PAM requirements allow researchers...discovered all the time and the requirements of these different systems can vary (for example, xCas9 is a variant...neither bind nor cut the target. Cas9 homologs from different organisms or Cas9 mutants developed in a variety...researchers to target many different genomic loci. 3. Cleave target DNA resulting in a double-strand break...). Throughout the remainder of this post, we’ll refer to this RNA complex as the “gRNA”. The gRNA contains...choice (see tips for designing a gRNA) and their prefered Cas9 variant (with the appropriate PAM sequence...sequence.   Now that you know about all the different components of CRISPR, It's time to start thinking...

...student or postdoc may require different types of mentoring. Some mentees prefer lots of advice and micromanagement...an effective working environment? Tom Ellis was offered a start-up package that included a postdoc and ...recommends to seek out advice from seasoned PIs who can offer support. “It is an exciting and emotional time!”...recruit people who are nice as top criterion and offers advice to not rush when growing your lab. “One ... lab and safety manager, researcher, adviser, conference planner, and writer.” Connie Cepko was surprised...impossible' or useless.” How do you run your lab differently or the same as your adviser did? Tom Ellis merges...online. The Howard Hughes Medical Institute (HHMI) offers some great resources for Early Career Scientists...

...check out these blog posts.       References and resources References Burry, Richard W. “Controls for ..., you will have a better understanding of the different types of controls and be able to use this information...treatment versus and which are likely just natural differences between individuals. Recommended controls for...extremely bright, then the noise likely will not interfere. If the fluorescence from your target is weak,...fluorescence of the sample, or autofluorescence, also interferes with data interpretation and can be addressed....  Western blotting When comparing samples in different lanes in a western blot, you’ll need to include...equal loading across samples and equal protein transfer from the gel to the membrane. A loading control...

...plasmid sample has a mix of different-sized multimers, you may even see different sets of bands on the gel... us at [email protected]. Resources and References References Bacolla A, Wang G, Jain A, Chuzhanova NA...Nanopore sequencing or PacBio SMRT sequencing, (which differ from the short-read Illumina MiSeq sequencing that...multimerization has been observed to occur in many different kinds of plasmids, taking many forms (Levene, ...from BiteSize Bio for more information on how different plasmid conformations run on an agarose gel). ...low metabolic burden seed-stock and fed-batch fermentation processes. Biotechnol Bioeng. 2009 Aug 15;103...Plasmids 101 eBookAddgene's Molecular Biology ReferenceAddgene's Restriction Digest Protocol More resources...

...libraries expand upon the CRISPR family of plasmids by offering a lentivirus-based mechanism for sgRNA delivery... microscope 8 hours post-pulse. The distinct differences of red-to-green fluorescence in individual mitochondria...mitochondria in the networks marks the differences in import of newly synthesized (green) protein and ...five previously unknown channelrhodopsins from different species: Stigleoclonium helveticum (ShChR), Chlamydomonas...advances, the Ting lab’s first-generation APEX suffered from inconsistent activity levels and limited ...cassette. They validated this methodology in two differents types of bacteria (E. coli and Salmonella typhimurium...naturally-occurring variations in the TALE repeat, referred to as “non-RVD variations”, were found to improve...

...isolated from different organisms and engineered Cas9 variants that were able to target different PAM sites...activity of Sniper-Cas9 to different Cas9 variants using gRNAs of different lengths (Lee et al., 2018)...found in other species can recognize a variety of different PAM sequences. Researchers have also used experimental...activity relative to Cas9. Since then, the Joung and Jennifer Doudna Labs used targeted mutagenesis to generate...relative to eSpCas9 and SpCas9-HF across a number of different sites. Using a combined random and directed mutagenesis...choosing a Cas protein for your experiment. References Hu, Johnny H., et al. "Evolved Cas9 variants ...

... the best method for titering AAV? Is there a difference in titer results between ddPCR and qPCR? A: At...curve and somewhat more reliable quantitation. Differences in sample treatment across methods can affect...  Q5: What is the efficiency of co-delivering different AAV particles into the same cells in vivo? A: ...give some insight into codelivery efficiency at different doses. See Figure 4 in this paper for more information...very high dosage. Any thoughts?  A: The question refers to this paper. AAV can be safely concentrated to...IACUC committee.  Animal Models Q9: Is there any difference in transfection efficiency when delivering these...injections? A: We have not observed any appreciable difference in AAV transduction efficiency between retro-...

... the retina using viral tools.   References and Resources References Pillay, S., et al. "An essential ...and can be specifically targeted to a number of different cell types even when systemically administered...researchers speak about AAVs, they are actually referring to rAAVs. ITRs and serotypes The names of AAVs...ITRs, which comes from the AAV2 serotype. Serotype refers to the type of capsid (outer protein shell) used...capsids allow the virus to infect (or not infect) different cell types with some specificity. There are a ...will lose expression quicker than terminally differentiated, non-dividing cells.   Figure 2: ...subretinal injections of neonatal mouse eyes at two different ages." Retinal Degenerative Diseases. Springer...