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...vectors. To quantify this comparison, capsids were packed with a GFP transgene and GFP intensity and transduction...versus AAV-PHP.eB in the CNS. A GFP transgene was packaged into either an AAV-PHP.B or an AAV.PHP.eB capsid...versus AAV.PHP.S in the PNS. A GFP transgene was packaged into either an AAV.9 or an AAV.PHP.S capsid. Both...here or check out these protocols if you want to package your own virus. ...

... plasmid sample enters Addgene the same way. A package is delivered by a mail courier and then the journey...    The plasmid's journey begins First, each package is opened and the contents are carefully inspected...deposits arrive from labs around the globe. The packages have spent days to weeks in transit and some have...barcode can store a lot of information in a small space and is ideal for tracking large collections. Many...

...variable chains are secreted into the periplasmic space to be oxidized and form disulfide bonds (Frenzel...does not, be sure to set the absorbance to 280 nm (Pace et al., 1995). You’ll also need to know the extinction...Aggregation and storage Aggregation of antibodies may impact their quality and efficacy (Wang et. al., 2018)...10.1002/bit.27776. Epub 2021 May 4. PMID: 33811659. Pace CN, Vajdos F, Fee L, Grimsley G, Gray T. How to ...

...lost during fixation with these chemicals, which impacts sample structure and could also cause your target...destroy it for others. That said, organic solvents’ impact on lipids means that they are able to permeabilize....   Figure 1: Choice of fixation method impacts antibody binding. Formaldehyde cross-links the ...detergent has different chemical properties that will impact the level of permeabilization and membrane integrity...

...individual presynaptic terminals, which are jam packed with everything needed for synaptic transmission...?  ExM provides a straightforward answer to the space problem. Instead of imaging smaller, what if we ...the anchoring, digestion, and expansion steps can impact the integrity of different targets and labels. ...continuity can appear reduced in ExM, which could impact how you interpret a target’s structure. For example...

...binding capacity and high protein retention. It comes in a variety of pore sizes and protein capacity, allowing...conjugate will have to be tested to ensure it doesn’t impact the antibody’s binding to your protein of interest... is a little quicker to use, with high binding capacity, though it has lower retention after binding and...10 minutes) and simple, utilizing individually packaged membrane ‘sandwiches’. They work well for proteins...

...lentiviral infection begins with your cells, both your packaging cells and your target cells. Poor tissue culture...harvest after transfecting with the transfer and packaging plasmids. A benefit of the multiple harvest strategy...benefitting your cells, medium exchanges may also impact the quality of the lentiviral particles you harvest...density, volume, incubation time, and MOI can all impact the success of your lentiviral infection. To ensure...

...the CNS by lentivirus, which has a larger packaging capacity than AAV. Such an approach was described ...should be screened for activity in vitro and then packaged into AAV for in vivo delivery. Multiplex gene ... also possible and is only constrained by AAV packaging limits (i.e., how many sgRNA expression cassettes...

...differing customs processes, and the distances the packages needed to travel meant the accessibility varied...such as using couriers that refill dry ice if the package stays in transit for a certain number of days and...distributor who, among other things, could ensure our packages navigated customs and arrived at their destination...for tools such as empty backbones, lentiviral packaging systems, and luciferase expression. On individual...

... target is present immediately upstream of a Protospacer Adjacent Motif (PAM), the sequence NGG.  If these... SpCas9 and its enhanced variants, eSpCas9 and HypaCas9, are great choices. Check out our SpCas9 containing...Cas9 variant with higher specificity (eSpCas9 and HypaCas9) instead.  Choosing a Cas9 for site-specific modifications... within the editing window (bases 4-8 of the protospacer). While this is a small window, this system can...

...complete package, all of the pieces of software that depend on each other for the entire package to run....in-house software has been crucial for us to keep pace with our current need to analyze nearly 300 sequences...sequencing data collection and analysis eventually outpaced our software, even with continuous updates. The...

....1231143  Gaudelli, N. M., Komor, A. C., Rees, H. A., Packer, M. S., Badran, A. H., Bryson, D. I., & Liu, D.... J. M., Lee, S.-J., Liquori, A. J., Murray, R., Packer, M. S., Rinaldi, C., Slaymaker, I. M., Yen, J.,...10.1038/s41587-020-0491-6  Komor, A. C., Kim, Y. B., Packer, M. S., Zuris, J. A., & Liu, D. R. (2016). Programmable...-S., Walton, R. T., Delano, J., Butcher, R. M., Pacesa, M., Smith, B. K., Christie, K. A., Ha, L. L., ...

...take up essentially all of an AAV’s ~4.5 kB packaging capacity. Luckily, researchers have devised methods...the chosen acceptor vector. The vectors are then packaged into AAV virions via transient transfection of...the viral vector. This delivery method frees up space in the AAV allowing a researcher to fill it with...

...many of the same restrictions, like limited cargo space (~4.7 kb) for your genetic payload — there are a... cell types not of interest. Systemic capsids packaging Cre-dependent transgenes can also be paired with...enhancers, and switches all take up precious cargo space in AAV vectors. Dual expression systems, which use...to be administered concurrently to minimize the impact of neutralizing antibodies developed in response...promoter/enhancer screening. However, AAV-PHP.AX packages viral genomes at a lower efficiency than AAV9 ...Zheng, Q., Kozareva, V., Beddow, T., Tobey, I. G., Pacouret, S., Chen, A. T., Chan, Y. A., Ryvkin, D., Gu,...

...color":"#0052a9","css":"#0052a9","hex":"#0052a9","opacity":100,"rgb":"rgb(0, 82, 169)","rgba":"rgba(0, 82..."css":"rgba(0, 82, 169, 75%)","hex":"#0052a9","opacity":75,"rgb":"rgb(0, 82, 169)","rgba":"rgba(0, 82,...color":"#ffffff","css":"#ffffff","hex":"#ffffff","opacity":100,"rgb":"rgb(255, 255, 255)","rgba":"rgba(255...–296. https://doi.org/10.1038/s41594-018-0028-6 Packer, M. S., & Liu, D. R. (2015). Methods for the directed...

...rich and diverse workforce. Out of all the career spaces I explored for my post-PhD life, the science policy...webinar via the NC Science Policy student group SciPolPack for a more detailed discussion on how to to define...to chat with someone working in a science policy space you’re interested in. Many fellowship programs can...her work focuses on incentivizing the real-world impacts of health services research. She is particularly...

...recombination between homologous pairs before viral packaging even begins. This spontaneous recombination persists...2020). In general, we observe that 0.1 - 0.8% of packaged viral genomes contain a recombined/functional ...which can lead to off-target expression of the packaged transgene in neurons and glia.   Check the recombination...v14715, titer 2.4x10e13 GC/mL) for this study were packaged at Addgene and had a relatively low recombination...

...lentiviral capsid-based bionanoparticle system for packaging Cas9/gRNA ribonucleoproteins for transient expression...aptamer which binds an aptamer binding protein for packaging into a lentivirus-like particle. Read the paper...paper in Nucleic Acids Research Find the gRNA and packaging plasmids   Cytosine deaminase for RNA editing...

...environmental impact on a scale of 1-10: the lower the score, the lower the environmental impact in terms ... product? How can we consider the environmental impact of those processes? Who even regulates them? You...

...this time more visible and thereby have a positive impact on science. ReFigure saves time, knowledge and ...definitely give you credit for it and increase your impact! Download ReFigure extension and visit ReFigure.org... with Joe Akin. She envisions a world where the impact of teaching and doing science is rapid, concrete...