...creative commons license.    The introduction The introduction serves two purposes. First, it should... to have conflict. There are a lot of ways to introduce conflict into your work, but one of my favorite... two other things to complete your story: an introduction, and a resolution.   Fig. 2: The structure...ever thought about how X works?”, or even by introducing a person that the audience can relate to. It ...read (or listen or watch) further.  Second, the introduction needs to provide enough information so that ...thing applies for science storytelling. You can introduce your results right at the beginning of your communication...Addgene blog SSCS: Cognitive Load SSCS: Preprints Intro to our Summer SciComm Series  ...

...26973 AAVrg pAAV-hSyn-hChR2(H134R)-EYFP   Control AAV Control AAV allow researchers to not only interpret...inventory. Our new AAVs include: Optogenetics AAV Control AAV Biosensor AAV And more! Optogenetics Optogenetics...more efficient tools for light mediated neuronal control and we are expanding our viral service to make ...own experiments. To this end, we offer specific control AAV in smaller aliquots of 20 ul for serotype testing... service has now expanded to offer many of our control plasmids in new serotypes. See our Serotype Testing...response to action potentials, is now offered under control of the CAG promoter for ubiquitous expression. ... 44362  AAV1 pAAV-hSyn-DIO-hM4D(Gi)-mCherry Controls Plasmid Serotype Name 50459 AAV1 pAAV-hSyn-DIO-mCherry...

...which introduces functional Cas9 protein and the targeting guide RNA into cells by electroporation. Results...of the nuclease dissociates very slowly after introducing a cut, with residence times on DNA of approximately...free to anneal to complementary ssDNA molecules in vitro. Figure 1: Cas9 binding to the cleaved DNA... if we could leverage this model (built from in vitro studies) to boost the efficiency of sequence replacement...enables sequence replacement without cutting Our in vitro results also demonstrated that catalytically inactive...advantage but error-prone repair of the gene would be disastrous (i.e. if breaking the gene is worse than leaving...homology-directed human genome engineering by controlled timing of CRISPR/Cas9 delivery." Elife 3 (2015...

...oviductal lumen, tweezer-type electrodes are placed to mediate electroporation. Ohtsuka et al. compared traditional... homology arms and a selection marker, then electroporate it into mouse embryonic stem cells. After selecting...from Yang et al. requires two gRNAs targeting two intronic regions and two repair templates in order to insert...promoter upstream of the left homology arm, perform in vitro transcription, and use the IVT product as a template...compatible with standard microinjection and electroporation methods using in mouse transgenics. Designing... allele requires you to target 2 loxP sites to intronic regions 0.5 - 0.8 kb apart. In the example below...between the ampulla and infundibulum. In vivo electroporation is performed immediately after injection. Image...

... of biogenesis and mitochondrial degradation in vitro and in vivo. Merged green and red channel fluorescence...Technology Austria, decided to take a novel approach to control for these effects, drawing on the rapidly growing...1):39-48. APEX2 for proteomic mapping and electron microscopy A thorough understanding of complex...enabled progress on both fronts – as a genetic electron microscopy (EM) tag providing superior subcellular...also available at Addgene and can be used as a control for your experiments. For more information, please... allowing labs to precisely and non-invasively control neuronal signaling. For more information on DREADDs...Chen and colleagues for genome editing inXenopus tropicalis. (Guo et al., Development. 2014..) A new, higher...

...is the best-case result. For example, you have introduced a GFP protein into your cells (visualized via...104–105 signal intensity. If your samples and controls were processed the same way, the GFP- population...population should overlap with the negative control population (Figure 1). Based on where the GFP- population... Example of yes-no flow plot of the (negative) control (left) or with the sample (right). Negative and...biorender.com. Pro tip! If your samples and control underwent different treatment processes, you may... population, so it no longer overlaps with the control. In tricky experiments, these populations may overlap...positions to the base expression. When compared to the control — which should be base expression — a peak to the...

...Lab and the Chang Lab have created methods that introduce diversification by targeting a site of interest...which limits the level of variation that can be introduced.   Diversification is limited due to the integration...use of such libraries means that the diversity introduced by lambda red recombineering and Multiplex Automated...PCR, EvolvR uses an error prone polymerase to introduce mutations. However, by using nCas9 to direct PolI3m...the 3' end of the nick (right), occasionally introducing mutations (red nucleotide). Created with BioRender.com...EvolvR, three additional amino acid changes were introduced to nCas9. These changes promote nCas9’s dissociation...increased the likelihood that a single gRNA would introduce more than one mutation near the target site. Applications...

...one type of virus to choose from (lentivirus, retrovirus, AAV, etc.). When you’ve decided on an application...experiment. Research applications Gene expression control Researchers are always wanting to manipulate gene...complementary mRNA sequence for degradation. Viral introduction of shRNA plasmids can induce gene-specific silencing...Viral CRISPR-Cas9 tools have been engineered to introduce frameshift mutations in genes of interest, turning...depending on the virus used. All of these expression control mechanisms require a specific modification of the...that the copies of library cargo delivered are controllable. For many screening applications, it is essential...attenuated, nef-deleted HIV-1 strains in vivo. Retrovirology. 2007 Sep 23;4:66. doi: 10.1186/1742-4690-4-...

...Photoactivatable Fluorescent Proteins Control Protein Expression with Cre-lox Control Protein Activity with GFP ...transmitted by intermediate messengers. Optical control of kinase activity can provide greater spatiotemporal...There is no common method for creating optically controlled kinases. The Solution: In Zhou et al, the Lin... the delayed effects because you can precisely control the timing of kinase activity. 2. Studying negative...photoswitchable kinases in vivo not only allows for precise control of a signaling pathway in the context of a whole...addition, it could be possible to use pdDronpa to control other kinases and proteins. Zhou et al suggest ...Pengpeng Li, Kang Shen, and Michael Z. Lin. "Optical control of cell signaling by single-chain photoswitchable...

...guides are generated by introduction of a single photocleavable 2-nitrobenzyl linker in the 20-bp targeting...nucleic acids or bridged nucleic acids can also be introduced in the gRNA. These modifications result in conformationally... innate immune response associated with gRNA introduction (Allen, et al., Frontiers in Genome Editing)...experiments in which viability is being compromised by introduction of foreign RNAs or immunity itself is being ...technologies utilize chemical modifications to the RNA to control the entire editing system with a simple LED light...al. Photoswitchable gRNAs for Spatiotemporally Controlled CRSIPR-Cas-based Genomic Regulation. ACS 6, 5... Susanna Stroik...