...be packaged in a variety of capsids. To date, at least 12 different capsids, or serotypes, have been identified...using AAV is the limited packaging capacity. Lentiviruses have capacities of 8–12 kb, while AAV only has ...libraries available at Addgene. The large packaging capacity and stable expression of lentiviral vectors make... AAV, with the genome from one serotype and the capsid of another. Researchers have used these psuedotypes...Interest in interrogating and mapping the genetic landscape of tissue-specific diseases has grown exponentially...

...antibodies, Anti-SARS-CoV-2 Nucleocapsid Protein [mBG17] and Anti-SARS-CoV-2 Nucleocapsid Protein [mBG86], target...target distinct epitopes on the SARS-CoV-2 nucelocapsid protein, one of the virus’s most abundant proteins...Figure 3: HeLa cells transduced with SARS-CoV-2 nucleocapsid protein (Plasmid #141391-LV [+N]) or membrane...) were immunoblotted against Anti-SARS-CoV-2 Nucleocapsid Protein antibodies #211756 (left panel) or #211757..., et al. (2021). Development of a SARS-CoV-2 nucleocapsid specific monoclonal antibody. Virology. 558,...-strand AAV (ssAAV). To maintain the packaging capacity and improve the specificity of ssAAV, a targeting...

...intuitive and I was making figures quickly. Inkscape Inkscape is the most similar to Adobe Illustrator out...and like Vectr, Inkscape uses vector graphics. There’s a steep learning curve for Inkscape, but there are... desktop use for Windows, Mac OS X, and Linux. Inkscape is versatile and has a wide selection of tools...lots of resources for learning the software. The Inkscape website has many tutorials and Lynda has a tutorial...

... compared to the wild-type, escaped neutralization. “That degree of escape from a very small number of...luminescence.     B.1.351 variant neutralization escape The lab saw that the B.1.351 variant was very resistant...what was surprising was the extent to which the escape appeared to be happening.”  In their assay, the...-YS, Lessells RJ, de Oliveira T, Sigal A (2021) Escape of SARS-CoV-2 501Y.V2 from neutralization by convalescent..., Balazs AB (2021) Multiple SARS-CoV-2 variants escape neutralization by vaccine-induced humoral immunity...

... AAV capsids (left) and transduction patterns (right).  Both AAV-PHP.eB and the AAV.CAP-B capsids are ...positive and negative selective pressures for novel capsids. In a continuation of that work, they have found...is true in both mice and nonhuman primates (for AAV.CAP-B10, for example).  Given that the liver is an ... derived from AAV9.  Of those shown here, only AAV.CAP-B10 causes brain wide expression, with no liver...

...to the type of capsid (outer protein shell) used to package an rAAV vector; these capsids allow the virus...AAV components      Cargo capacity Empty rAAV has a cargo capacity of ~ 4.7 kb, meaning that your... virus, packaged in a non-enveloped icosahedral capsid, that can be used to express genes of interest ...plasmid and individually packaged into the AAV capsids. As a genome conversion in the transduced cell ...

...select novel AAV capsid variants could unintentionally limit the utility of candidate capsids.  Are you interested...small randomized peptide loop inserted in the AAV9 capsid. This mutant library was then screened in vivo ...version of LY6A, but a mutation in PHP.B’s unique capsid loop disrupted binding. Collectively these results...LY6A, a novel cellular receptor for AAV-PHP. B capsids." bioRxiv (2019): 538421. If you’re interested ...administration of the adeno-associated virus-PHP. B capsid fails to upregulate transduction efficiency in ...

... determine if CRISPR/Cas9 escape occurs in an animal model. If these escape mutations are frequent, the...their results, Wang et al. showed that HIV-1 could escape from CRISPR/Cas9-induced modification targeting...either the LTRs or essential genes. Many of these escape mutations were located near the Cas9 cut site, ...1 infection, which would make it easier for an “escape” virus to propagate and infect adjacent cells. ... Inhibit HIV-1 Replication and Accelerate Viral Escape. Cell Rep. 2016 Apr 5. PubMed PMID: 27068471. Ramanan...

... for the identification of plasmid multimers. Capillary Gel Electrophoresis (CGE) analysis Another technique...technique that can detect multimers is capillary gel electrophoresis (CGE), which has been used in plasmid...Thompson L. Vaccine Plasmid Topology Monitoring by Capillary Gel Electrophoresis. Curr Mol Med. 2020;20(10)...PMC4480603. Holovics HJ, He Y, Lacher NA, Ruesch MN. Capillary gel electrophoresis with laser-induced fluorescence...fluorescence of plasmid DNA in untreated capillary. Electrophoresis. 2010 Jul;31(14):2436-41. doi: 10.1002/...study and topology analysis of plasmid DNA by capillary gel electrophoresis. BioProcess Int. 2006 Sep;...

...your experiments. Capture MS Strategies This approach uses immunoprecipitation to capture proteins from a...and then uses mass spectrometry to identify the captured proteins. If the antibody is specific, then you...approach is ideal for validating antibodies for immunocapture applications and is amenable to high-throughput...to validate that your antibody is specifically capturing your RNA-binding protein of interest. So you ...immunoprecipitation with your antibody and send the captured proteins off for mass spec analysis (above). When...

...my training, I dove straight into my capstone project. A capstone project is a major project given to ...Figure 1: Addgene's summer interns presenting their capstone projects during the Summer Intern Poster Presentation... scientist. But I expected something wrong. My capstone project was based on plasmid cloning and I, the... Once everything was confirmed, I worked on my capstone project report and presentation, during the final...

...distinct behaviors such as spontaneous swimming and escape reflex. These traits, coupled with short generation...zebrafish at 5 days post fertilization, including escape behavior and spontaneous swimming. When startled...zebrafish react by performing a distinct C-bend to escape possible danger (see figure). Larvae also perform...in response to any specific stimulus. While the escape response neural pathway is well characterized, ... kind of swimming behavior, spontaneous swim or escape, was elicited when these different neuron groups...

...interactions. ChIP works by using antibodies to capture protein:DNA complexes with antibodies specific ...your protein(s) of interest. Once the complex is captured, it is treated so that the protein and DNA are...DNA and then determine concentration.  Protein Capture After crosslinking and sonication or enzymatic ...antibodies, with your samples, and then use an antibody capture method such as Protein A/G beads to collect your...and the Three Steps (Crosslinking, Fragmenting, Capturing), if you ask me.  Each of the three steps need...

...the potential hazards of creating “gene drives” capable of spreading through wild populations. Whereas ...organism carrying a synthetic gene drive were to escape the laboratory, the drive could eventually spread...protocols, they can reduce the risk of accidental escape to a negligible level. One major reason we chose...chose to publish the concept and likely capabilities of RNA-guided gene drives in the summer of 2014 (after...performing experiments in a geographic area where escaped organisms won't be able to find mates2. For example...

...polymerase (reviewed in 2, 5). The minigenome is capable of being replicated and transcribed in the cell... Minigenome System). However, minigenomes are incapable of producing infectious virus (similar to lentiviral...VP30, and L (reviewed in 5). The RNA genome is encapsidated by the nucleoprotein, NP. NP associates with...mRNAs are translated) by cellular machinery. NP encapsidates the negative sense, single-stranded RNA minigenome...viral polymerase complex then recognizes the encapsidated minigenome as a template for replication and...

...Transgenes That Exceed the Packaging Capacity of Adeno-Associated Virus Capsids. Human Gene Therapy Methods 27...make it a great viral vector, but its packaging capacity is limited to ~4.7kb, or roughly half the packaging...Lastly, depending on its design, donor A may be capable of expressing a truncated protein. This can be ...Expands Adeno-associated Viral Vector Packaging Capacity in a Transgene-independent Manner. Molecular Therapy...

...methods termed proximity-based labeling to better capture these interactions in a vast array of biological... of biotin for avidin, proximity labelling can capture weak/transient protein interactions as well as ...exposed lysines on neighboring proteins. Thus to capture protein complexes, scientists can simply express...After a few hours biotinylated proteins can be captured on streptavidin affinity beads or matrices. The...transpeptidase sortase A variant (mgSrtA), which is capable of promiscuously labeling various cell surface ...

...of working with fruit flies is their ability to escape their habits and live in the lab itself. Many of...of tiny black dots during our stay there. Yet capturing the flies is nearly impossible, due to their small...everywhere find and re-contain any specimens that escape during routine lab activities.  Here, we demonstrate...plasmid-based system for locating and containing escaped flies. Nature 4:1-22. https://doi.org/4.01.2022...

... and migrate within neurons, and that are also capable of spreading across synaptic connections. Most ... like lentivirus, but instead is contained by a capsid which is comprised of the viral proteins VP1, VP2...genetically engineered to have differences in their capsid proteins. For a brief overview see Addgene’s AAV...relatively small (20 nm) in size which makes them capable of diffusing in the extracellular space; they also...include AAV1 – 9, with most of these serotypes capable of entering neurons (see table 1)(Choi et al., ...2017 also provide evidence that AAV1 and AAV9 are capable of synaptic spread. The underlying mechanisms of...

...editors CRISPR toolkit for epitope tagging Lentiviral capsid-based Cas9/gRNA ribonucleoprotein delivery Cytosine...protein-truncation strategy with reduced RNA-recognition capabilities.  Read the paper in Nature Biotechnology Find...Cas9/gRNA ribonucleoprotein delivery by lentiviral capsid-based bionanoparticle Transient expression of CRISPR...Recently, Baisong Lu’s lab developed a lentiviral capsid-based bionanoparticle system for packaging Cas9...