... Viral Vector Packaging Service Viral Production Viral Production Jump...cells using the 2nd generation psPAX2 and pMD2.G packaging system developed by the Trono lab . Cell culture...

...the genetic code for reverse transcriptase are packaged (Wu et al., 1997) inside of the capsid . This ...the vector particle, some of the RT protein gets packaged inside of the vector particle, along with its ...-type viral genome can direct the production, packaging, and release of progeny viral particles. And there...4355596 King, S. R., Duggal, N. K., Ndongmo, C. B., Pacut, C., & Telesnitsky, A. (2008). Pseudodiploid Genome... Temin, H. M. (1990). Genetic consequences of packaging two RNA genomes in one retroviral particle: pseudodiploidy...mouse embryo cell lines in which each cell has the capacity to be activated to produce infectious murine leukemia...

...action at a distance.” Through the marvel of DNA compaction, an enhancer can increase the expression of a...millions of base pairs away. (For details on DNA compaction and the structural proteins that make it possible...details. DNA is a 3D Fractal Figure 2: DNA compaction. Graphics courtesy of beyondthedish (left) and...tertiary structures (globules) to achieve exquisite compaction—on the order of 700 terabytes per gram—and form...chromosomes we all know and love. To understand compaction, suppose you have a piece string that’s 10 meters..., are mere millimeters apart. Similarly, cells compact DNA in a way that brings linearly distant regions...

...Gairik Sachdeva. Healthcare consulting is a fast-paced field, requiring people who are willing to quickly...uncertainties and a rapidly evolving healthcare space. The consultants who enjoy their jobs most, get ...typically cutting edge; a consultant might outline the impacts of combining targeted molecular therapies for cancer...consulting can sometimes be challenging. The fast paced nature of projects often means working long hours...more. Essentially any role in the broad healthcare space that can benefit from the blend of business and ...interested in the business of life sciences and it's impact on human health.     Resources on the Addgene Blog...

...single-strand AAV (ssAAV). To maintain the packaging capacity and improve the specificity of ssAAV, a targeting...CRISPRi screen to identify additional host factors impacting prime editing and found the protein La (a small...proteins and a critical component for viral genome packaging. Addgene recommends both for use in western blots...that was more desirable than having many closely-packed labeled cells. Plus, Voltron2’s fluorescence is...vectors for microglia was further enhanced by packaging the virus as self-complementary AAV (scAAV). However...

...system by modifying paCas9-1 using nMagC714 instead of nMagHigh1-C714, generating paCas9-2. This change did...VVD, N. crassa) (Kawano et al., 2015). Nicknamed paCas9-1 and consisting of the fusion proteins N713-pMag...fold-induction of Cas9 activity (16.4-fold). This paCas9-1 light-inducible system was able to recognize ...(flCas9). When triggered by blue light (470nm), paCas9-1 induced indel mutations via NHEJ (frequency of...their prior work, the Sato lab also showed that the paCas9-2 system could be spatially controlled and reversibly... of gene transcription Nihongaki et al., 2015 paCas9 Magnet proteins from N. crassa Split Cas9, Split...

...hazards for these reagents. Work in a well-ventilated space and under a fume hood when working with the volatile...

...architecture of my cells,” or “How does this mutation impact the types of cells found in my tissue.” It is based...antibody performs differently and some antibodies impact the performance of other antibodies. But, once ...the right protocol, multiplex IF can give you impactful and beautiful results. Performing an immunofluorescence...fixation maintains sample morphology with minimal impact to the target epitope. There are multiple methods...exclusive to IF, but they can have a significant impact on the outcome of your IF experiment. Many of the...choose for your experiment will have a significant impact on the protocol you use and the ultimate outcome...

...will be moving into a bigger space so that we can make an even bigger impact - we are always on the lookout...Can you imagine how much space a million plasmids would fill? Two weeks ago, Addgene edged passed our...assembling the boxes, to inoculating tubes, and to packing and shipping. Every single day. 673 plasmids shipped... hard to picture one million plasmids. How much space would one million bacterial stabs take up? If 81...

...Construction and Evaluation Accessory Pack Takashi Yamamoto This accessory pack contains modified pFUS array vectors...

... to scientists around the world. To support this pace of research, new, centralized databases and centers...protocols, publications, and more to accelerate the pace of research into COVID-19. While this program is...response initiatives. Accelerate the translational impact of research Universities have also been stepping...research synergies and accelerating the translational impact of research activities. By streamlining the development...by coming together, we can create a significant impact on both clinical care and public health for this...

...to revisit some of the ways these animals have impacted humans through biomedical research. The Camelid...Camelid family is composed of camels, llamas, and alpacas (and their undomesticated counterparts) - not exactly...soon after found similar antibodies in llamas and alpacas (Figure 1). At the time, researchers were already...started with immunizing a camelid (usually llamas or alpacas). But because sdAbs are so amenable to recombinant...And I suspect we will only continue to see their impact grow as new sdAbs become easier to generate and...

...bioRxiv. AAV2 interacts with the packaging cell's cell membrane When packaging adeno-associated viral vectors... vector production. Whether redesigning viral packaging plasmids, investigating different viral production...heparan sulfate proteoglycans on the surface of the packaging cells (Vandenberghe, 2010). Rather than being ...prevent the uptake of AAV2 viral particles by our packaging cells by treating the cells with the endocytosis...

...Viruses are simple: a genome packaged in a protein shell (Taylor, 2014). They’re so simple that we can...simple, small particles have quite the outsized impact — and not just on the disease front. Viruses survive...particles. As a bonus, this increases the carrying capacity of the viral vector, increasing both safety and..., just to increase the viral vector’s carrying capacity. In order to allow the viral vectors to reproduce...necessary, genes for the required proteins are packaged in separate plasmids and expressed via cells cultured...

...realizing it. Yet, this selection is crucial as it will impact downstream outcomes of an experiment, including...Grimm & Lavis, 2022). These variations directly impact the way you can utilize these dyes. The most-used...emission of labeled agonists and antagonists, impacting the interpretation of experimental results. It's...dopamine D2/D3 receptors (Allikalt et al., 2020) Impact of labeling process: The process of dye labeling... on the respective fluorochrome. Brightness is impacted by both the excitation coefficient, characterizing...multiple dyes during analysis. Image resolution is impacted by several factors. Firstly, brighter dyes enable...

...expression at varying capacities. AAV vectors containing the AAV2 ITRs can be cross-packaged with capsids from...replication and are required for replication, packaging, and vector persistence. They work in conjunction...in the same DNA molecule as the gene that they package. In a transfer plasmid, the transgene is flanked...Bonnin, D., Merten, O.-W., & Galibert, L. (2017). Impact of Inverted Terminal Repeat Integrity on rAAV8 ...

...developed the Phage-Assisted Continuous Evolution (PACE) and Phage-Assisted Non-Continuous Evolution (PANCE...mutations, selection, and replications per day.  The PACE and PANCE systems use M13 bacteriophage, which rely...dilution step represents the major difference between PACE and PANCE: for PANCE the phage progeny is not diluted...-stringency PANCE, followed by higher stringency PACE, resulted in the evolution of TadA variants with...contexts. References Gaudelli NM, Komor AC, Rees HA, Packer MS, Badran AH, Bryson DI, Liu DR (2017) Programmable...

...psPAX2  - psPAX2 is a 2nd generation lentiviral packaging plasmid from the Didier Trono lab that expresses...MuLV retroviral particles. It can be used with packaging plasmids such as pCMV-dR8.2 dvpr (lentiviral) ...pCMV-VSV-G. pRSV-Rev - This 3rd generation lentiviral packaging plasmid from the Trono lab contains the rev gene...pMDLg/pRRE - This is a 3rd generation lentiviral packaging plasmid from the Trono lab that contains the gag...-puro contains all the necessary elements for packaging, reverse transcription, and integration needed...

...Expression Infects Non-Dividing Cells Large Packaging Capacity (>8kb) High Transduction Efficiency Transient...for Mouse Cells and Hematopoietic Cells Low Packaging Capacity (~4.5kb From basic science to clinical...accommodate a cargo up to approximately 8 kb, but viral packaging is less efficient when inserts are on the higher...

...nontargeting spacer flanked by two CRISPR repeats. The spacer can be digested using BsaI, allowing a spacer-repeat...using 3 distinct spacer-repeats, you could easily create 7 unique arrays (e.g. for spacers A, B, and C, you... cassettes, making them advantageous for small capacity vectors like AAV. In addition to the mammalian...