...specific, programmable platform well-suited for various innovative biological and translative technologies. Addgene...only deactivate the Type I-F system and are not translatable to the other CRISPR subtypes. Still, the large...Mali et al. in January 2013 (7, 8). Nine months later, more than 1500 publications describe work to improve...enabling access to multiple plasmids used for various platform applications. For more information on CRISPR/Cas...

...pAAV.hSyn-FLEX.iGABASnFR.F102G) is noticeably dimmer than iGABASnFR, and accumulates in the endoplasmic reticulum, but exhibits larger...GlyR IRES EGFP Chimeric channels for neuronal manipulation 119742  AAV5  AAV SYN PSAM4 GlyR IRES EGFP...EGFP Chimeric channels for neuronal manipulation Recombinases Plasmid Serotype Name 50363  AAV5 .... It is useful for activity imaging of large populations of densely labeled neurons because background...  It is useful for activity imaging of large populations of densely-labeled neurons because background...

...with regard to compliance with health authority regulations,” Baradaran says.  Baradaran’s path to science...is an Addgene depositor. KSQ uses a CRISPR-Cas9 platform to systematically test the function of all 20,... a genetic screen to identify the factors that regulate cell division. And that really got me hooked on...lentiviral microRNA-based system for polymerase II-regulated RNA interference. In the first two months since...also interested to see Addgene expand the sharing platform to other reagents in biology. Luk H. Vandenberghe...

...establish a next-generation sequencing (NGS) QC platform that allows us to provide scientists with complete...launching our viral service in 2016, we adapted this platform to accommodate adeno-associated viral vector (...ligation and tagmentation require a double-stranded template.  Using viral genome sequencing to verify the ...double-stranded species that could be used as a template for downstream applications (Berns, 1972). Using...the Addgene blog Read all of our viral vectors related blog posts Read our blog post on AAV titers Browse...

... of the time because of relationships. It’s much harder to build relationships when you are more desperate...your hand at a video presentation? Most video platforms make it easy to record yourself. You can watch...experiences to share. The conversation builds a relationship for the future and the 1:1 format is good for...you will make good progress on both building relationships and thinking about future career options. 5....

...activity of two populations of neurons at once. Addgene spoke to Boyden about this latest optogenetics ...compete against each other. Getting two-color stimulation has been a holy grail of the field. The problem...selective for different ions. Right now, we can modulate electricity. If you could selectively let in calcium...Independent Optical Excitation of Distinct Neural Populations. Nathan Klapoetke et al. Nature Methods. 2014...

...switched on and off reversibly. This ability to modulate fluorescence allows scientists to observe single... (T203H) (1). wtGFP normally contains a mixed population of neutral (protonated) and anionic (deprotonated...T203H mutation in PA-GFP generates a chromophore population that is more neutral, causing PA-GFP to display...with blue light (1). Similar to PA-GFP, scientists later developed red fluorescent PA-FPs by performing several...photons achievable with photoswitchable dyes. Correlative light and electron microscopy (CLEM) is an imaging...Ryoko, Hideaki Mizuno, and Atsushi Miyawaki. "Regulated fast nucleocytoplasmic shuttling observed by reversible...

...site-specific recombinases, regulatory elements, developmental regulators, and lots of viral components... as Pichia pastoris). If your goal is protein isolation, then the MoClo Pichia Kit will get you started...developed to study the basic biology of and gene regulation in the opportunistic pathogen Acinetobacter baumannii... Molecular Tools to Study Gene Expression and Regulation in Acinetobacter Baumannii.” ACS Synthetic Biology.... “A Unified Multi-Kingdom Golden Gate Cloning Platform.” Scientific Reports, vol. 9, no. 1, Springer ...

...1, to create a “cumulative wealth distribution” curve, or Lorenz curve. By calculating the area under ...can be counteracted by growing the cells on solid plates, where each transformant forms its own colony, ...less of a concern, but Addgene still recommends plating the transformation mixture on solid media. The ...The colonies are then scraped off the plates and pelleted for DNA extraction. Prepare library product for...

...vector production, assembled AAV vector particles accumulate inside the producer cells and purification protocols...gradients. The current purification protocols for isolating exo-AAVs rely on differential centrifugation steps...Figure 2). Since the development of the exo-AAV isolation protocol by Maguire and colleagues, other researchers...et al., 2017), and exo-AAV8 expressing human coagulation factor IX in liver in a mouse model of Haemophilia...et al. 2018). The structure of exosome AAVs is related to the use of microvesicles as a vehicle for drug...

...engineers regularly use the platform, why scientists are using the platform, and tips for getting the most...from Nature Jobs Blog While Twitter is a popular platform for scientists, it’s not the only tool in a scientist... Facebook. For each of the three social media platforms, there’s some statistics on what percent of scientists...given you tips for using various social media platforms, and 3) helped you fit social media into your ...

... on the properties found in the literature or tabulated data that summarizes properties of fluorescent...) respectively. The theoretical brightness is calculated by multiplying EC by QY (EC*QY, sometimes normalized...several important experimental conditions that are related to the imaging strategy and the sample. Practical...developed in our lab uses a reference protein that is translated from the same open-reading frame and separated...the assay is depicted in figure 1. The strict correlation between the fluorescent protein of interest and...

...splitting up a sample into thousands of physically isolated partitions (such as droplets or wells on a chip...edit. Since exploitation of NHEJ to generate translational reading frame-altering indels for “functional...wild type target in each sample can be quickly calculated (details in [2]) and can be used to quantify ...few thousand cells from single wells of a 96-well plate without prior quantification. Comparatively, mismatch...AnotherLabRat.     References 1. Miyaoka, Y. et al. Isolation of single-base genome-edited human iPS cells without...

...Antibodies, 1999). This creates an immortal cell population that will continue to produce the specific antibody...or, are injected with the whole protein. After isolating the B cells from an immunized animal, those B ...step. Then, a limiting dilution is performed to isolate individual hybridomas and ELISA is used to identify...the selected hybridoma is expanded as a clonal population (Holzlöhner et al., 2017). Why choose monoclonal... Addgene’s antibody plasmid collection Read the latest on Addgene's new antibody project ...

...gamma-retrovirus is Murine Leukemia virus (MLV). Naturally isolated gamma-retroviruses have been associated with cancer...lentivirus and gamma-retrovirus are so closely related, their required components are very similar. While...heterologous envelope or a post-transcriptional regulatory element can be used for either type of retrovirus...infect humans, many mammals, and even the avian population. Some adenoviruses are known to cause respiratory...induce a lower inflammatory response within hosts. Relative to the other viruses discussed in this article...

...mouse lines, you can access and manipulate an even further defined population of neurons. For example, say... workhorse adeno associated virus (AAV) shows relatively low retrograde transport efficiency in most serotypes...expression, detecting neuronal signaling, or manipulating any other genetically controlled function. By...Fig 3B). These specific neurons could then be manipulated (e.g., activated or inhibited) by any genetic...

... promoter, gRNA and pol III terminator. These relatively large cassettes (considering the gRNA itself ... vector. In addition, the pol III promoter is relatively weak, and low expression of gRNAs from these ...deletions of other regions, such as noncoding RNAs or regulatory elements, to examine their function. Bacteria... of polycistronic gene organization, and its translation to CRISPR/Cas9 shows great potential for multiplexed...gRNA Multiplexing Multiplexed and programmable regulation of gene networks with an integrated RNA and CRISPR...

...aren’t, so repeated cycles will favor accumulation of the latter products. As noted above, a single cycle...repair, as there is no requirement for a repair template (sister chromatid or homologue) or extensive DNA... repair should take less than an hour, thus a population of cells constitutively expressing a targeted...NHEJ-dependent repair (“pop-in” insertion) provided a template containing compatible overhangs is available. ...

...watch my webinar, “Not Networking 101-Building Relationships for Success”. Choose 1-2 tactics to try. One...be a bit random because you never know which relationships will take off, or who your new contact will ...current organization but also important to build relationships outside your organization. The mentor in your... doing it again next month?” Some mentoring relationships grow naturally out of friendships or work interactions...other ways they can help you grow. Mentoring relationships can last for an hour or for a year. Some click...

... at a relatively low frequency, and in nondividing cells, this pathway is further downregulated. Rather...the cell to use the deaminated DNA strand as a template. To do so, the lab used a Cas nickase, instead...repairs the DNA using the U-containing strand as a template, copying the base edit. Cytidine deamination...repair, thus removing these cells from the edited population.  Another way to improve base editing efficiency...While cytosine base editors often produce a mixed population of edits, ABEs do not display significant A to...