We narrowed to 820 results for: APP

...simpler, yet more effective, CRISPR-based epigenetic approach (spoiler alert: it’s dCas9… with no modifications...). But there are a number of drawbacks to this approach: TET proteins are not demethylases; they are ...and Szyf 2021).    Figure 3: A simpler approach - dCas9 blocks DNMT1 methylation, leaving the ...resistance genes.  It’s that simple! Good luck and happy editing!    Thank you to our guest blogger!Daniel...Addgene blog CRISPR101: Editing the Epigenome  Mapping the 4D Nucleome with CRISPR/Cas9 Resources at ...Xu X, Tao Y, Gao X et al. (2016) A CRISPR-based approach for targeted DNA demethylation. Cell Discov 2,...

... Ophthalmology Basel recently took a different approach to enhance specificity in retinal cells. To do...type specific expression in the retina.  Four approaches for designing synthetic promoters  To build a...reproducibly labeled retinal cells can be found here.  Applications for synthetic promoter AAVs Besides restricting...synthetic promoters have a few other potential applications: Using promoter combinations for targeted expression... promoters expressed in. The lab tested this application using one promoter to drive expression of Cre-mCherry... rates of labeling are ideal of gene therapy applications, promoters that produce sparse labeling would...

... to increase AAV transduction efficiency by approximately 15-fold (Chen et al., 2006). 3. Transduction... at 4 °C). After ultracentrifugation, virus may appear as a white pellet. The pellet can then be rinsed... This antibiotic selection can be performed approximately 72 hours after transfection and should give ...give rise to a surviving population of approximately 20–50% of the packaging cells after 1–3 days of selection... could be examined with a microscope and should appear fluorescent if they were infected. While there ...transfection, higher-level expression after approximately 72 hours is likely the result of successful ...

...to be less expensive than an ELISA kit. In this approach, viral RNA is first converted to cDNA and then...several different lentiviruses in parallel, this approach should be considered. Similar to the p24 assay... Fluoresence Titering Protocol Another popular approach utilizes the antibiotic resistance genes carried...have integrated into a target cell’s genome. This approach can discriminate multiple integration events, ...underestimating the viral titer. The main drawback of this approach is that it can be quite labor intensive. Target...virus may want to choose more straightforward approaches such as the FACS-based or colony-forming unit...

...your resume pulled to the top of the pile when you apply for such positions. Of course, if your goal is to...transition from an academic setting—especially if job applications require teaching experience or high numbers ...web site. Each position is for a specific, pre-approved project. Other companies, such as Novartis, have...s program. Once you’ve found a few positions to apply for, compare and contrast industry postdocs at those...not. The point is that each company has a unique approach, and some might suit your needs better than others...background in cell and developmental biology and is applying this towards researching neurodegenerative diseases...

...10 modules often rely on two-step hierarchical approaches using different Type IIS restriction enzyme specificities...transformation plate can be manipulated to result in appropriate levels of colony plating densities. Ligase fidelity...accuracy of assembly. The fidelity data can be applied to derive similar high-fidelity overhangs for any...researchers continue to test the limits of DNA assembly approaches with increasingly complex experimental conditions...Kucera is a Principal Development Scientist of Applications & Product Development focusing on DNA cloning...Optimization of Golden Gate assembly through application of ligation sequence-dependent fidelity and bias...

...18% to 65% (Maggio et al., 2014). An alternative approach to NHEJ is to utilize the Homology Directed Repair...phenomenon to the presence of terminal proteins capping the ends of linear double-stranded AdV genomes....double-stranded chromosomal DNA breaks “that always happen naturally.” These adenoviral CRISPR/Cas9 genome...and quiescent, post-mitotic mammalian cells. Applications of CRISPR delivery by adenoviral vectors Since... Adv vector delivery is often used, the CRISPR approach based on HDR has the advantage of being a potentially...Strategies for Tissue-Specific Targeting. In: Applications of viruses for cancer therapy. Elsevier, pp ...

...The strategy they landed on was to recombinantly append their proteins of interest with unrelated peptides...generate a reliable antibody for protein tagging approaches” was not part of the goal when Evan’s team generated...plays in human health and disease. But no science happens in a vacuum!  Munro and Pelham learned of these... flexible - it could now be used in way more applications.   Fig. 1 Making a Protein Tag (Abridged...their study, some of the newer Myc-tag antibodies appeared to be less sensitive to sequence context.    So...proteins expressed from cloned genes: deletion mapping functional domains of Drosophila hsp 70. Embo J...

...and Mar. 1, 2020. Figure 2: A simple approximation of the effect of COVID-19 on data smoothed with...a 7-day rolling mean. While this global approximation already shows us a rough "COVID-19 effect" on...maximum likelihood estimates of each parameter. We approximated the data likelihood with multivariate Gauss-...the supposed end of lockdown), request activity appears to return to normal. The model fit these two discrete...is a Ph.D. candidate at MIT. His research uses applied machine learning and statistical methods for fundamental...fundamental scientific discovery and high-impact applications.       Alvin Hsu is a graduate student at Harvard...

... and are ready to move on to your downstream application — but wait! Do you know how much plasmid DNA ...reading results from a NanoDrop. The principles will apply to similar instruments from other companies, like...sample will have minimal impact on downstream applications, but they can impair your ability to get an ...be sure it is compatible with your downstream applications and that you use the same solution for blanking...are, of course, more considerations and other approaches for measuring DNA quantity and quality than we...A230 ratios in RNA preparations on downstream applications. Qiagen Gene Expression Newsletter. Issue 15...

...involves partitioning a PCR reaction mixture into approximately 20,000 droplets using water-oil emulsion technology...to create a water-in-oil emulsion containing approximately 20,000 droplets. Each droplet contains the material...droplets. In the image to the right, there is approximately 1 positive copy per microliter in the no template... use ddPCR for AAV titration, there are many applications for the technology.  Droplet digital PCR is ...being used in the detection of microbes. Some applications include measuring viable probiotics and detecting...detecting circulating pathogens.  With its many applications and ease of use, droplet digital PCR is becoming...

...a number of labs are using enChIP, and I'm very happy to see that many researchers all over the world ...describe this technology and some of its potential applications? Fujii: We generated a series of retroviral ...the peer-review system of journals and grant applications, for example. However, at the same time, it'...Throughout your career, you have held research appointments in the US (New York University School of Medicine...which only one was done in Japan, which is very disappointing. I hope I can help change the situation in the...and graduate students as they complete their appointments in your lab?  Fujii: These days it is very difficult...

...their experimental needs, and some of the common applications in the lab. Why Green Fluorescent Protein?...Re-engineering GFP to increase its range of color and application Almost as soon as its sequence was elucidated...the mutation definitions found in Table 2 may not apply. Table 1: The specific mutations comprising common...others) Neutral mutations    A multitude of applications Due to its size and ease of use, GFP and other...change. Typically, two fluorescent proteins with overlapping excitation/emission spectra are used; one fused... just scratched the surface of the potential applications for GFP. It has also been used to identify particular...

...performs well in many different antibody-based applications.  IPI’s recombinant version is a fusion of mouse...ligase cereblon (CRBN), including popular new approaches like PROTAC targeted protein degraders. But CRBN.... elegans embryo, where developmental processes happen within a few hours. In order to use the latest ...cellular processes in C. elegans: mScarlet3 for applications needing intense brightness (such as low-expression...simplicity and efficiency of this dual-plasmid approach makes it a valuable resource for advancing our...when the module unfolds under tension. When this happens, BiT and the LgBiT luciferase fragment (expressed...

...difficult to source a commercially available and appropriately conjugated antibody. While labs can conjugate...find,  it is often easier for labs to source appropriate antibodies for an indirect ELISA than for a direct... capture antibody must bind to distinct, non-overlapping epitopes so that they do not interfere with each...specific and can be difficult to develop if an appropriate conjugated antibody is not readily available....cross-reactivityChallenging to develop assay Competitive Appropriate for small moleculesReduced chance of cross-reactivity...choose the best ELISA for your needs. Good luck and happy binding! More resources on the Addgene blog Antibodies...

...these proteins are quite large — comprising approximately 1000–1400 amino acids — which poses a challenge...-strand breaks like those created by Cas12a. Applications of TnpB One of the major challenges in genome...), which is especially valuable for regulatory approval and public acceptance. Recently, viral vectors... is where TnpB offers a clear advantage. At approximately one-third the size of Cas9 or Cas12a, TnpB is...viral vectors. Even for advanced genome editing approaches like base editing and prime editing (Molla et...viral vectors and for multiplexed gene editing applications. As research into TnpB continues to evolve, ...

...This distinct approach means researchers can now select the best tool for their application from among many...incorporate A bases across from abasic sites. This approach resulted in a maximum A → T product purity of ...AYBEv3!). Although AYBE, AXBE, and ACBE do not approach the high product purity of CBEs and ABEs, they...). Several different teams succeeded with this approach around the same time (He et al., 2024; Tong et...on in the base editing field! From CRISPR-free approaches to mitochondrial base editing to base editors...

...hour. Find the SHERLOCK plasmids at Addgene! Applications of the SHERLOCK detection system The Zhang lab...portability. This method can be used for an array of applications including clinical diagnostics (e.g. pathogen...While the lab points out that the protocol is not approved for clinical use at this point, they hope that...patient samples. Find the DETECTR plasmids here! Applications of DETECTR in diagnostics DETECTR allows for...an array of molecular and clinical diagnostic applications. The company Mammoth Biosciences has been started...SHERLOCK detection system, DETECTR has not been approved for diagnostics yet, but is undergoing validation...

...cells retain the same color, allowing for the fate mapping of cell populations over time. The ability to track...variety of species, cell types, and research applications. Labeling neurons with Brainbow The Lichtman...neurons in the cerebellum of mice, the team appropriately named their technology Brainbow. Brainbow 1.0...level of these XFP is not strong enough for all applications. To resolve this, similar systems have been ...cell type when crossed with mice expressing the appropriate Cre allele. The Confetti mouse contains the Brainbow...PMC3713494. Ghigo, Clément, et al. "Multicolor fate mapping of Langerhans cell homeostasis." Journal of Experimental...

...Depending on the research question and downstream application, one or several of those techniques can be used...your flow analysis. However, certain downstream applications, like an in vivo adoptive transfer of those ...or free amines of proteins. Depending on the application, they can be membrane-permeant or membrane-impermeant...are positive or negative for the dye. The main applications of dye stains in flow cytometry are cell proliferation...with a high resolution for specific downstream applications, while maintaining a clean cell surface, dump...and bind DNA or free amines, depending on the application. Paul Heisig is a Research Associate in the lab...

...AAV11 ✓ ✓ AAV1 AAV1 was the first viral vector approved for gene therapy (Issa et al., 2023). It may be... 2016). This functionality makes it useful for mapping neural connections. Although derived from AAV2,...vessel barriers. It is especially popular for applications targeting liver tissue. In addition, it has ...the blood-brain barrier, making it useful for applications targeting the CNS (Issa et al., 2023). Additionally...treatment, which can pose problems for gene therapy applications. Cross-reactivity of AAV serotypes varies by.... A. (2023). Various AAV Serotypes and Their Applications in Gene Therapy: An Overview. Cells, 12(5), ...

...you with these, so keep reading! Good luck, and happy optimizing!  Recommended Reading Issa, S. S., Shaimardanova.... A. (2023). Various AAV Serotypes and Their Applications in Gene Therapy: An Overview. Cells, 12(5), ...A. (2021). Next Step in Gene Delivery: Modern Approaches and Further Perspectives of AAV Tropism Modification...AAV-Mediated Anterograde Transsynaptic Tagging: Mapping Corticocollicular Input-Defined Neural Pathways...Retrograde Serotype Viral Vectors 101: Viral Applications...

...secondary antibodies.   Application-specific controls Some antibody-based applications have very specific ...then moving on to controls for antibody-related applications. After reading, you will have a better understanding... unavailable, try substituting with knockdown approaches using siRNA or shRNA. If this alternative is ... alternative, albeit less ideal, verification approaches discussed in the Antibodies 101: Validation post...Secondary antibody specificity  In indirect staining approaches, secondary antibody specificity must also be ...

...making it very useful for genetic engineering approaches. Most importantly for this technology, however...piggyBac™ technology can be used for numerous applications, including transgenesis, gene-trap screens, ...ssODN) as the donor [6]. The problem with this approach is that, since there is no selection, you will...technology that may be used for a variety of research applications, including making transgenic cell lines and ..., while avoiding the pitfalls common to other approaches. Many thanks to our Guest Bloggers Amar Singh...Fingers, TALENs, and CRISPR-Cas9 nucleases. Mathew applies his specialized knowledge to help researchers identify...

...created by the Recipient Right to file patent applications on modifications or inventions Right to publish...can even sign electronically or choose to auto-approve MTAs. Whatever your preference, we are here to ...to make it fast and easy. Fast Approval: The UBMTA used by Addgene is internationally recognized as...transfer officers are familiar with the UBMTA and are happy to agree to it. Their confidence and familiarity...familiarity with the UBMTA result in quick approvals, so your plasmid requests can get to your lab faster. For...

...have used fluorescent proteins to observe what’s happening inside cells for at least a couple of decades....of hues. While those tools are great for many applications, Verkhusha and his lab at Albert Einstein College...exactly what they reported doing in a paper that appeared in Nature Biotechnology back in 2011 and then ...Verkhusha’s team reported another advance in the application of near-infrared fluorescent proteins, iRFP670...contrast agents for two-color imaging in animals. The approach relies on ultrasound waves as opposed to light...

... Normal Variables – Independent Variance – Approximately equal Data type - Quantitative If your data ...null hypotheses. Benjamini uses a complementary approach which identifies a p value cut off. This method...all generally yield a similar result and are appropriate to use if all sample comparison is needed, without...keep your friendly neighborhood biostatistician happy by making your statistical analysis plan when you...right statistical test, just do your homework and happy testing!   References Wissing, Dennis R., and Timm...Variables: A Monte Carlo Simulation Study. Front. Appl. Math. Stat. Volume 2 (2016). doi.org/10.3389/fams...

...increase specificity. This modular approach made ZFNs highly appealing to researchers.  TALENs In 2010, ...fiction, was also a trained biologist. This is apparent in his work The Island of Dr. Moreau (1896) where...made SciFi director Ridley Scott wonder what would happen if these corporations would extrapolate genetic...successful in the box office, indicating the insatiable appetite of the moviegoers for dinosaurs wreaking havoc...also transcended into the world of SciFi. This is apparent in the use of the word ‘CRISPR’ for anything associated...

...thermophiles, bacteria must be grown at lower temps. This approach only works for facultative thermophiles (high ... occurs? This is different than the engineered approach used to make eSpCas9 and SpCas9-HF1. By mining...SpCas9 RNP’s and a guide targeting a site with an overlapping PAM. In both cases, GeoCas9 and SpyCas had similar...could also be used with in vitro molecular bio applications that require high temperature reaction conditions...always, the plasmids are available from Addgene. Happy CRISPRing! References Harrington, Lucas, et al....

...beneficial for use in therapeutic or behavioral applications where drawn out effects are required. Targeted... subcellular regions of a neuron. Optogenetic approaches excel at discrete cell or tissue-subregion targeting...to the limits of light delivery. Chemogenetic approaches excel at broad tissue or system-wide targeting...chemogenetics. Note that, as with all tools, both approaches suffer from unique sets of off-target side-effects...EndogenousNot easily controlled Unique advantages Mapping the brainDissecting circuits Studying GPCR pathways...