...same result – uncontrolled movement of the vehicle or, in the case of cancer, uncontrolled cell proliferation... cellular level there is a universal theme - uncontrolled cellular proliferation. We have appreciated ...appreciated for some time the causes of this uncontrolled proliferation, in particular the roles of proto-oncogenes...opposite task, keeping cells from dividing out of control. One may compare the roles of proto-oncogenes and...

...of all distributed plasmids as part of quality control, but does not perform functional tests. The newest...produced from a plasmid in cells. Additionally, in vitro assembly of NgAgo/ssDNA requires incubation at 55...can conveniently transfect/inject cells with in vitro preassembled Cas9/gRNA complexes or with guide RNA...does however rely on cloning into a vector or in vitro synthesis, which could be less desirable if one ...et al. "DNA-guided genome editing using the Natronobacterium gregoryi Argonaute." Nature biotechnology ...

...drug-resistant population compared to a non-treated control group. In this scenario, gRNAs that are “enriched...future therapeutic targets for diseases causing uncontrolled cell growth, such as cancer. Figure 2...As with any experiment, the use of appropriate controls, multiple replicates and several cell types can...of multiple gRNAs targeting the same gene can be strong evidence that a particular gene is actually important...Titering Your Lentivirus Plasmids 101: NGS Quality Control for Pooled Libraries Find More CRISPR Resources...

... create a single polycistronic 15 kb insert. The final construct was then introduced to HeLa cells (Figure...resulting HeLa cells show robust labeling, with strong expression and detection of each fluorophore at...meaning that (in the previous example) one can control whether the third module is added to the 5’ terminus...degree of identity, so not suited to generate polycistronic containing multiple identical polyadenylation...

...diseases such as hemophilia, muscular dystrophy, spinal muscular atrophy, neurodegenerative diseases and metabolic...life-threatening (cystic fibrosis, hemophilia, muscular dystrophy)? AAV gene therapy started in the mid-90's with...larger transgenes. Unfortunately it has limited tropism in the retina, primarily infecting cells of the...epithelium (RPE). AAVs, on the other hand, have broad tropism in the eye. AAV-2, the first identified serotype...

...techniques involve the assembly of a circular plasmid in vitro, before transforming it into E. coli for propagation...specifically cuts methylated DNA, so it will selectively destroy the template, but not newly synthesized sequences...protein-coding sequence, or a mutation can be introduced in a gene, while adding a GFP-tag (Figure 3)....unamplifiable’ Check your amplification by gel electrophoresis. In our experience, good amplification after...interested in neuronal signal transduction, and uses electron cryo-microscopy to understand receptor function...

...now many variations, including many that use auxotrophic rescue genes as reporters and some that repress...example of the former. In this system, histidine-auxotrophic host strains are used to select for binding-dependent...accounted for using appropriate replicates and controls (see Tips below), and by validating leads with...reporter gene activation. Including a prey-only control also provides a baseline level of reporter activation...that are known to bind serves as a good positive control to ensure that your setup works. The last thing...

...generated a series of retroviral vectors expressing tagged dCas9 and guide RNA. The retroviral enChIP system ...requested plasmids for enChIP through Addgene. Retroviral Expression System for enChIP using CRISPR Addgene...Addgene: Your lab recently developed a retroviral enChIP system using CRISPR technology. Can you describe...Associated with an IFNgamma-Responsive Promoter by a Retroviral Expression System for enChIP Using CRISPR. Fujita...

...research. Many other mutations have since been introduced to GFP and new iterations of fluorophores are... Transcription Reporter: Placing GFP under the control of a promoter of interest can be used to effectively...Scientists can use light to detect, measure, and control molecular signals, cells, and groups of cells in...Plasmids of this type may have the GFP under the control of an additional promoter from that of the gene...fate studies. It can also be used, when put under control of promoters of interest, to visualize the developmental...

...organisms to control their movements and behavior. These functions and others are controlled by collections...tomography, super-resolution STORM imaging, and electron microscopy. Sample preparation for these techniques... et al. "High-performance probes for light and electron microscopy." Nature methods 12.6 (2015): 568. ...the Addgene Blog Rabies and neuronal tracing Retrograde AAV: Making the journey from axon to nucleus ...