...optimum of 30 °C and is therefore inefficient in mammalian cells (usually grown at 37 °C). Smart molecular...populations) within neuronal networks in living animals. To accomplish such cell specific manipulations.... “Conditional Site-Specific Recombination in Mammalian Cells Using a Ligand-Dependent Chimeric Cre Recombinase...

...COVID-19 (Joung, Ladha et al., 2020). Cas13a in mammalian cells The Zhang lab made the exciting discovery...discovery that some Cas13a orthologs can function in mammalian and plant cells (Abudayyeh et al., 2017). They...superfolder GFP fusion to stabilize the protein in mammalian cells. LwaCas13a-msfGFP can mediate cleavage with...

...reagents and process. It is important to note that optimal experimental conditions, including how much plasmid...backbone as Plasmid A—that does not target any mammalian gene. This non-targeting control is critical to...a gene, you should design a control that shows maximal activation. Likewise if you are trying to repress...

...pool down to plasmids expressing variants with optimal function. High-throughput DNA sequencing is then...ubiquitination activity assays, toxin activity, small molecule binding, growth rate, and many more. See..., Gibson assembly is better suited for creating small, focused scanning libraries. PFunkel: Of prior...

...Approach: Viral RNA is extracted and subject to isothermal amplification by primers that target the E and...RNA is extracted from a sample and subject to isothermal recombinase polymerase amplification (RPA) using...contain reverse transcriptase to produce cDNA. Isothermal amplification is done at one constant temperature...

...Good fixation maintains sample morphology with minimal impact to the target epitope. There are multiple...Sample Preparation This step can involve multiple smaller steps that all help to ensure that antibodies can...Sample Preparation, this step can involve multiple smaller steps, but the aim is to preserve sample integrity...

...deliver the best possible videos for free. We’re a small operation, so sometimes it can be a big challenge...premium, so I had to figure out a solution for that. Normally, shots require me to get in close proximity to...good once. Even when things return to relative normalcy, I’ll be incorporating that mobile monitor stand...

... refer to antibodies and 'staining' to refer to small molecule dyes. Indirect labeling While antibodies...of those CD8+ T cells, require the cells to be minimally altered, so you would not be able to use antibodies... Lastly, stains, based not on antibodies but on small molecule dyes, are useful, most notably when it ...

...The new SuperPLDs work with high efficiency in mammalian cells and in vitro, opening up a wide realm of...directed-evolution methods, the activity-based selection in mammalian cells used here could be an attractive strategy...Activity-based directed evolution of a membrane editor in mammalian cells. Nature Chemistry, 15(7), 7. https://doi.org...

...science, and can include: equipment, reagents, animals, glassware, and even people. When developing a ...budget bottom line costs may be rather high. That’s normal, because this is where you can start to identify... modeling and collaborations rather than using animals, then that’s a cost that you can eliminate from...

... will be able to provide letters of reference, informal recommendations, more positive grant application... not know.... 6. Go to science meetings big and small Meetings give you and your research exposure. Use...Eat well- a diet rich in vegetables, foods with minimal processing, and low in empty calories will make...

...little tissue absorption, weak tissue scattering and small amounts of tissue auto-fluorescence (Drobizhev et...excitation wavelength as it has been shown that even a small incremental change of the excitation wavelength ...Dynamic imaging of dendritic cell extension into the small bowel lumen in response to epithelial cell TLR engagement...

...Function Antigen presenting cells (APCs) that would normally alert the rest of the immune system to the presence...and IL-10 that suppress APCs. T cells that would normally attack the cancer cells are also suppressed by...activity of CTLs and NK cells. These cells would normally kill cancer cells but are directed not to through...

...choice, and size of the target brain region, the optimal viral titer must be determined empirically (this... weeks over 8 weeks) in order to determine the optimal time period between viral injection and commencement... of neural circuits underlying brain disease in animal models. Nature reviews Neuroscience. 2012;13(4)...

...5], Sendai virus [6], plasmid electroporation (episomal) [7, 8] and mRNA transfection [9]. Many of the...differentiation.” Specific cell media, supplements, bioactive small molecules, and growth factors are used to control... While natural transdifferentiation is rare in mammals, an example is observed in the pancreas when excess...

...lentiviral vector to be used in mammalian cells, check Lentiviral and Mammalian Expression). This information...differing in the gene/insert. Pooled libraries are normally supplied as liquid DNA with all of the plasmids...

...Differential cytosine editing between bacteria and mammalian cells Researchers in the Xueli Zhang lab were ...seemed to follow the same initial mechanism as in mammalian cells — by excising the freshly edited U bases...found that C → G editing was often accompanied by small deletions in their target loci (between 4.0% to ...

...gave to me: a miniature CRISPR-Cas system for mammalian use. On the second day of CRISPR, Addgene gave...Addgene gave to me: MUCICAT to perform multicopy chromosomal integration.  On the tenth day of CRISPR, Addgene...

...which are used to deliver Cas9 and other genes to mammalian cells. For this reason, the top requested plasmid...envelope protein under the control of the powerful mammalian CMV promoter. Envelope proteins are a key component...

...editing and multiplex prime editing (requiring minimal payloads!) Uncovering Ancestral Bias in CRISPR ...Miles on everything from AAV prep to targeting to animal models. It was one of our favorite posts from 2022...