We narrowed to 820 results for: APP

...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), ...

...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 ...

...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...

...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...

...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...

... 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...

...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...

...users have recently begun to develop CRISPR-based approaches for studying gene function in these systems. ...use in a fetal-lung derived organoid system, the approach is intended to be amenable to organoids derived...tissue-types. By bringing together CRISPR-based approaches and organoid models, toolkits like this one provide...trigger seed germination of parasitic plants, this approach was also used to build and validate a ratiometric...parasitization of cereal crops. As a result, agricultural applications of these novel biosensors could have a major...

...scientific aspects of writing. Following his advice, I applied to the class. Receiving the email that I had been...the ASM headquarters. I decided to go ahead and apply; thinking I probably would not get selected for ...knowledge with practical understanding of how to apply it. The personal interaction of critiquing others...well as being critiqued added an extra level of application not present in the online class.   Four ways ... manuscript should be able to be enjoyed and appreciated by all fields not solely the specific area the...

...tag to remain on your protein for downstream applications Below, we describe a few of the most common...not to perturb protein function for downstream applications. Tag cleavage must be performed after primary...also provides a visual detection mechanism via applications such as western blot and immunofluorescence....dimerizes in solution, which can affect downstream applications. Also, purifying GST by affinity chromatography... are generally best suited for their primary application. Ready to start tagging? We are here to help ...

...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....

...learn how easy this process is and to get tips for applying it in your own lab. Table 1: Comparison Between... E. coli and transformants selected with the appropriate antibiotic. Although the precise mechanistic ...additional 15-17 bases that are complementary to the overlapping vector ends. See an overview of the PCR process...an agarose gel. You should see bands with the appropriate sizes for your vector and insert. Once you’ve...correct product. Once you find a colony with the appropriately sized product, purify and sequence the plasmid...

...isolating the cell line all the way to modern applications. What the HEK is it?   HEK cells were first...cell biology, there are thankfully many other applications that they are useful for! What the HEK is it...cell line and we will highlight some of those applications here.   Virus production HEKs are the go-to ...proteins at high levels. In a case where clinical applications of recombinant proteins are necessary, HEKs ...significantly reduces the cost of production. Research applications There are two areas of research where HEK cells...

...NgAgo replace CRISPR? Such a drastic shift won’t happen overnight, but there are a few reasons why you ...flexibility in choosing a genomic target. NgAgo also appears to outperform Cas9 in GC-rich regions. DNA guides...staggered (Cpf1) cleavage. NgAgo takes a different approach, randomly removing 1-20 nucleotides from the cleavage...site specified by the gDNA. Depending on your application, this could be favorable or unfavorable. In a...activity. One-guide faithful NgAgo guide loading appears limited to the period when the protein is being...

...generation of various flux imbalances. The conventional approach has typically been to either systematically delete...networks and multiplex gene control. However, these approaches are often met with the challenges of regulatory... possible. From a design perspective, this is appealing since the RNA scaffold modules are comparatively... multiplex control via scRNA design is readily apparent in metabolic engineering. The authors elegantly...control of gene expression. This strategy can be applied to effectively change metabolic flux. The dCas9...

...IVT sgRNA) approach (Dewari et al. 2018 and Figure 2). Not only is the cr:tracrRNA approach highly efficient...biochemical tasks in the cell. One of the most popular approaches to study protein interactions is the pull-down...disrupts protein function, an N-terminus tagging approach should be pursued, however, clonal lines must ...me at [email protected]. Keep pipetting and happy genome editing! Pooran Dewari is a postdoc in Steve...optimizing CRISPR/Cas9 genome-editing methods and applying them to study brain tumour biology. When not pipetting...

...axial dimension that will appear in focus). This means that the images will appear sharp without interference...excite a molecule of GFP. Because red light has approximately half the energy of blue light, two photons of...density of red-photons be establish for this to happen.Using red light has a second advantage: red light ...objects lying within the diffraction limit will appear as one object in the final image. Over the past...of light – think of a single GFP molecule – will appear as a fuzzy 200-300 nm spot when imaged. Super-resolution...

...interactions. Beyond these common applications, researchers are finding new ways to apply fluorescent proteins everyday...Zhou and Fatima Enam. There were many excellent applicants but Xin and Fatima stood out for their creativity...Light-inducible Proteins (FLiPs) Fluorescent proteins can be applied in a long list of creative ways. In her graduate...utility of these FLiPs in a variety of research applications and particularly notes their ability to be precisely...

... of applications with the potential to save lives. While most scientists recognize and appreciate the ... Yamanaka himself, during his keynote address, appeared almost incredulous that the field has exploded...daunting because they have no clear cut answers, I applaud the stem cell community for addressing these important...but they are also the reason this research is happening. Stem cells scientists have certainly not lost...

...have high levels of off-target effects, both of happen with Cas13. CRISPR-Csm degrades both nuclear and...antibodies may just not perform as well in one application as another, so it is always best to have options...for mEGFP-HRas (from Plasmid 18662). See the Applications section on the Anti-GFP [N86/8R] page for more...fluorescent proteins (FPs) necessary for many of these applications have lagged. Fixation, chemical staining, and...well-suited for the demands of advanced microscopy applications such as super resolution, electron, and expansion...

...altering its function). HDR is crucial for certain applications, for example, correcting the allele that causes...this question led us to develop a new, rational approach to improve the efficiency of sequence replacement...introducing a cut, with residence times on DNA of approximately 6 hours. To our surprise, we measured identical...region allowed sequence replacement rates of approximately 1%. This is by no means a large number, but ... order of magnitude greater than transfection approaches. Moreover, the cost of reagents is relatively...

...together a grant application? Are you familiar with the publishing or patent application processes? How ...How can you stand out among all the candidates applying for the position?   Here are a few things to consider...makes sense for your skills, and the job you are applying for. Also remember that a resume is typically ...have done is closely related to the job you are applying for, in which case talk about your research, as...

...next-gen sequencing. ChIP is an incredibly useful application for exploring these interactions, and can give...including epigenetic modifications. But it's not an approach for the faint of heart!  The Process ChIP can...preferred method, enzymatic digestion can be appropriate when your protein of interest is a low abundance... upfront investment than many other antibody applications, this investment can well be worth it once you... idea to add "patience and an extra-methodical approach" to the top of your protocols when embarking on...

... by the Dueber Lab at Berkeley, provides a new approach for generating novel genetic variants in bacteria...the key components of EvolvR and its potential applications. Challenges of existing mutagenesis methods...exist, but there are a few drawbacks with these approaches that makes it challenging for users to achieve...is capable of mutagenesis in E. Coli, but this approach requires special bacterial growth conditions in...more than one mutation near the target site. Applications of EvolvR The Dueber Lab used EvolvR to discover...

...multicolored nano-lanterns for use in various applications, including optogenetics, biosensors, and fusion...furimazine. In addition to the previously described applications, the enhanced signal of eNL allows their use...characterizing Nano-lanterns and eNL for many different applications. For long-term imaging, especially where photobleaching...concern, these constructs come in very handy. This approach is especially suited to complement optogenetic...Therefore, especially thick samples may not be appropriate for use with Nano-lantern technology. Multicolor...

...principles we used in our daily content writing applied to alt text. We also were aware of the need for...they hoping to get from it? The image in Figure 1 appears on our protocol page for AAV Purification by Iodixanol...a lot of jargon and have a technical, detailed approach. Other times, it is more helpful to use plain ...start small with alt text or immediately dive into applying these four steps elsewhere, engaging in this practice...from posters to papers.  We hope this helps, and happy writing!   Resources Additional resources on the...

...disciplines. In biology, innovative new imaging approaches are constantly being developed which combine ...biological protocols and revolutionary physical approaches. However, with each new technical achievement... greatly increases accessibility. Through this approach, the Planktoscope, a portable device for studying...on-going creativity with the construction plans. Appropriate open-source licenses for hardware (e.g. CERN ...optical setups. References Joel T. Collins, Joe Knapper, Julian Stirling, et. al, "Robotic microscopy for...

...advances and how some of these variants may be appropriate for your specific research question. Considerations...off-target activity, which may be undesirable in many applications. At the time of our first blog post on choosing...Using a combined random and directed mutagenesis approach, the Anna Cereseto Lab created pX-evoCas9, which...enzymes may better suit specific experimental approaches. One example is CasX, characterized by the Oakes...While no enzyme is perfect for all experimental approaches, knowledge of both the advantages and limitations...

...regions, both through genetic and proteolytic approaches. This comes in handy when, for example, you want... as secondary antibodies, have much broader applicability. It is fairly easy to find these in a Fab fragment...pre-form primary-secondary antibody complexes before applying them to samples, such as when performing IHC in...can be used as a block in similar scenarios. By applying unconjugated Fab fragments that are the same species...fragments described above can be used for similar applications and have similar advantages, though with some...

...two posts in this series described a practical approach to selecting a bright fluorescent protein and ...aggregates. This property can be tolerated in some applications (e.g. labeling of cells or tracking promotor...promotor activity), but it is problematic in applications in which the fluorescent protein is used as an inert...cells. To understand how well fluorescent proteins approach this ideal situation, cellular assays have been...can be used in the OSER assay including CytERM-mKOkappa, CytERM-mTurquoise2, and CytERM-dTomato. When...

...DNA-binding proteins to label certain loci, but this approach is not scalable for every locus...unlike CRISPR... 9 and 13. They next turned their attention to mapping pairs of intrachromosomal loci. The technique successfully...authors’ knowledge, this work represents the first mapping of intrachromosomal loci, a major benchmark in ...-MS2-PP7 Despite the success of Ma et al.’s approach, fluorescently labeled Cas9’s come with a number...up fluorescent CRISPR labeling, they took a new approach: labeling the gRNA itself. In their CRISPRainbow...

...some reporters are better suited for particular applications. Read on to learn more about each reporter or...sensitive than HRP RANbody detection, it is approximately as sensitive as detecting the same antigen with...less sensitive than HRP RANbody detection but approximately as sensitive as antigen detection with a standard...detected with a labeled anti-HA or -MYC antibody, approximately as sensitive as detecting antigen with a standard...Detection with an anti-Chicken IgY antibody is approximately as sensitive as detecting antigen with standard...

...brains. However, it is not known why or how this happens. Therefore, I set out to study how protein turnover...protein turnover in situ in neurons. The classic approach for studying protein turnover is radioactive pulse-chase...immunocytochemistry is perhaps the most common approach. With a good and specific antibody and the proper...proteins with FUNCAT Professor Schuman’s technique applies metabolic labeling using a noncanonical amino acid... observed with fluorescence microscopy. This application of AHA labeling is known as fluorescence noncanonical...

...engineering fields. We’ll walk you through how to apply this precise and easy-to-use system to your cloning...designed correctly, the recognition sites do not appear in the final construct, allowing for precise, scarless...are eliminated by digestion/ligation and do not appear in the final construct. The destination vector ...adapted Golden Gate cloning for inserting the appropriate oligonucleotides specifying a gRNA target sequence...different combinations, or other new and exciting applications, this system is an incredibly powerful tool ...

... editing in primary human cells  Cell therapy approaches can use base editors to correct mutations in ... reduced off-target editing frequencies. For applications that require high DNA editing specificity, for... for example for potential therapeutic approaches, the authors recommend mRNA delivery, the V106W version...deamination, an important attribute for therapeutic application. ABE8s demonstrate an overall improved base editing...editors with increased activity and therapeutic application. Nature Biotechnology. https://doi.org/10.1038...

...When possible, the PE3b/PE5b approach is recommended over the PE3/PE5 approach. Figure 7: Nicking...its design changes for every new prime editing application. And optimizing its sequence is important, as...with BioRender.com. PE3b and PE5b: In this approach, the nicking sgRNA is designed to bind only after...BioRender.com. MMR inhibitors PE4 and PE5: These approaches inhibit MMR to improve editing efficiency, but...

...information to Addgene Material Transfer Agreement (MTA) approval  Send plasmids to Addgene  Quality control  In...can also directly paste sequence data into the appropriate full and partial sequence cells. *Note: If you...descriptive plasmid name and match the name as it appears in its publication or as it is commonly known in...descriptive purpose focuses on the practical application of the plasmid and will help a scientist outside... account username. In order for the plasmids to appear in your Addgene account under My Plasmids, we need...

..., its dimeric nature can interfere with some applications. Several labs have been working to produce monomeric... for molecular fusion and membrane-targeting applications. Nature Methods, 1–9. https://doi.org/10.1038...successful in primates. The authors used a multispecies approach to generate and screen a library of AAV9 variants...proven itself a good option for essentially all applications. Addgene’s plasmid collection includes over ...clone (M2) has been widely used in a variety of applications. Find anti-DYKDDDDK recombinant antibodies here...