...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...

...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...

... 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...

...degrade any residual DNA carried over from the packaging process. Incubate at 37 °C for 45 min. You are...

...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 ...

...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...

...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...

...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...

... systems are made up of separate envelope and packaging plasmids as well as transfer plasmids. This concept...in our Gamma-retrovirus guide and find popular packaging and gamma-retroviral envelope plasmids here. Lentiviral...can be found in our Lentiviral guide. Popular packaging and lentiviral envelope plasmids are listed here...gamma-retroviruses use the same gene products for packaging, however, the isoforms of these proteins, as well...differ. Therefore, lentiviral and gamma-retroviral packaging plasmids are not interchangeable. General envelope...

...in the compacted region and results in a loss of gene expression. In this way, genes in compacted regions... is a complex of DNA and histone proteins that compacts cellular DNA into chromosomes and reduces the ...and tightly bound DNA is to histones, the more compacted it becomes. This prevents the cellular machinery...study epigenetics. This is likely to have huge impacts in the way we study and treat complex or multifactorial...

...enzymes prefer targets with a given single base protospacer flanking site (PFS) sequence, but orthologs like...be made when multiple As are present in the gRNA spacer, as ADAR will preferentially edit an adenine if...PspCas13b-ADAR2DD(E488Q) displayed robust editing with various spacer lengths from 30-84 nucleotides, and designated ...they confirmed A->I editing and found that 50 nt spacers increase editing efficiency but also increase off-target...various dRanCas13b truncations that would allow packaging the construct for viral delivery. They found C-terminal...

...commonly associated with CRISPR arrays and involved in spacer acquisition following infection, as a form of “...is reduced when 2 mismatches are present. The protospacer flanking sequence (PFS) for LshCas13a, which ...sequence for Cas9, is located at the 3’ end of the spacer sequence and consists of a single A, U, or C base...each DNA target have high sequence identity to the spacer sequence and contain the appropriate PAM sequence...cells. LwaCas13a-msfGFP can mediate cleavage with spacers from 20-28 nucleotides in length and does not require...

...investigators realize too late that they have missed the impact of dark, non-fluorescent, and misfolded FP-fusions...quantitative imaging experiments or even negatively impact cells (see Figure 1). When behaving properly, ... the function of resident ER chaperones and may impact cellular function or possibly viability. FP misfolding.... It remains unclear whether the FP misfolding impacts fusion protein function and this could lead to ...secretory pathway (Figure 2), the inner membrane space of mitochondria and chloroplasts, the extracellular...

...aspects of research at National Aeronautics and Space Agency, the Environmental Protection Agency, and...long-term science outreach, may have the most profound impact (15). Diversity and inclusion in science, technology... immigrants, and minorities who have had great impacts on STEM (21–26). Increased scientific funding...communities (something already encouraged by “Broader Impacts” requirements for certain grants) (28). As one ...Science Foundation 15-008: Perspectives on Broader Impacts. The Pipettepen: The March for Science Raises Concerns...