...the HIV-1 genome had been excised, leaving only a small portion of the LTRs joined together. Clonal Cas9...preclinical HIV-1 research that should be pursued in animal models, and it’s difficult to tell if one approach...both cases, translating the in vitro work into an animal model comes with two key hurdles. The first is ...genomes will need to be sequenced to determine the optimal gRNAs for either “shock and kill” or viral excision...to determine if CRISPR/Cas9 escape occurs in an animal model. If these escape mutations are frequent, ...

...designing your plasmid – you carefully selected the optimal promoter for your gene of interest, painstakingly...coli are mainly found in the intestinal tract of animals. There are many different naturally occurring strains...arabinose as a carbon source leuB β-isopropyl malate dehydrogenase inactivated Requires exogenous leucine...is non-functional. If a gene is deleted, it is normally noted with a Greek delta (Δ).  Additional resources...of Replication The Promoter Region – Let's Go! Mammalian Vectors Looking for More Plasmids 101 Articles...

...terminals of axons form connections to dendrites of proximal neuronal cells. In the specific example of episodic..., pressure injection or iontophoresis (applying small electric currents via inserted electrodes) may be...clearer picture of neuronal connectivity in the mammalian brain (Wickersham et al., 2007; plasmids at Addgene...Addgene’s AAV guide. AAV particles are relatively small (20 nm) in size which makes them capable of diffusing...If Cre-dependent vectors are used in transgenic animals that express Cre in only specific cell types, for... titration on the Addgene website).  Tissue Optimal Serotype CNS AAV1, AAV2, AAV4, AAV5, AAV8, ...

...bacterial origins, Tet systems function well in mammalian cells, and TRE-containing promoters can be used...OFF. The cumate switch system functions well in mammalian cells, and can be used in the repressible manner...previous post. While originally developed for use in mammalian cells, this system has also been implemented in...Bujard H. Tight control of gene expression in mammalian cells by tetracycline-responsive promoters. Proc...-switch: a system for regulated expression in mammalian cells. BMC Biotechnol 6, 43 (2006). https://doi.org...

...allow.” Wang’s library is designed to target a smaller set of genes and it includes more guides per gene...sgRNAs, and can result in a higher titer due to the smaller size of the plasmids. The Zhang lab, on the other...possible to conduct systematic genetic screens in mammalian cells,” David Sabatini, a senior author of one...). Genome-Wide Recessive Genetic Screening in Mammalian Cells with a Lentiviral CRISPR-Guide RNA Library...

...plasmids are small circular pieces of DNA that replicate independently from the host's chromosomal DNA. They... modify, or construct a plasmid. A plasmid is a small circular piece of DNA found in bacterial cells, ...designed to introduce foreign DNA into another cell. Minimally, lab-created plasmids have an origin of replication...

...Then at your preferred time points, sacrifice the animals and fix the samples. When you use AAVs, it typically...typically takes a few weeks to achieve optimal expression of XFPs. Any longer and the cells may start to...to show morphological abnormalities due to the excessive amount of XFPs expressed. Then, clear the samples...suppressing transcription. It is critical to express a minimal amount of tTA to achieve highest expression levels...

...interested in studying animals than plants, and biology teachers’ preference for using animal systems to demonstrate...challenges than does working with animals. Because Arabidopsis is small, has a short life cycle, and is ...

...and serotype did you use and what animal subject did you use? For animal subjects it is helpful to report...experimental details (e.g. titer, injection site, animal strain used). Similar to the goodwill of Addgene... Immunohistochemistry? Did you use any relevant animal treatment conditions like drugs (Chemogenetics)...

...scicomm efforts fall under informal learning; however, many others partner with formal learning to engage students...peers can and do consume science communication.  Formal education is education that is provided through...fully and freely engage or disengage at will.  Informal education is education that is provided, typically...

...basis of transduction Transductions utilize the ‘normal’ mechanism of viral infiltration into a cell. The...be integrated into the host genome? Expressed episomally long-term? Expressed short-term? Answering these...non-integrating (adenovirus and AAV) but can still express episomally long-term (AAV). If you perform a lipid-based...transposon system, CRISPR targeted strategy, etc.) and episomal expression will last ~2-7 days at moderate levels...

...or other affected people, man/animal vs nature, anthropomorphized animals, or even seemingly unrelated ...Summer SciComm Series! We hope you've enjoyed this small glimpse into the world of science communication ...these pieces aren't always appropriate for your formal scientific communications, they are fantastic fodder...

...cellular dynamics with light by modifying EB1. EB1 normally sits on microtubules to attract other proteins... can be used to visualize RNA localization in mammalian cells. These tools consist of cobalamin-binding...in vivo. These new “Riboglow” probes contain a smaller RNA tag, are less prone to photobleaching, and ...your own experiments, mCarmine is available in a mammalian expression vector (pcDNA3) and a bacterial expression...

...Latin for 'it'), as “a generic term for any extrachromsomal hereditary determinant”. His proposal, however...1960s when scientists began to study other extrachromosomal particles, particularly Resistance or R-factors... deli in 1972. Using a deli napkin for paper, a small group of scientists including Stanley Falkow, Stanley...genes from different bacterial (and eventually mammalian) species were cloned into plasmids and new cloning...

...most of this new relationship? Consider adding formality and active goal setting to your mentoring relationships... Check out Joanne's Reddit AMA Add a little formality to mentoring Just getting together and chatting...phone, etc.)? Make agendas for your meetings, take informal minutes on what was covered and learned, especially...do after I get my PhD) and break them down into smaller, actionable steps (e.g. Meet 10 PhDs with interesting...

...Pasteur, Paris) that changed genome editing in the malaria community for the better (Nat. Biotechnol., 2014...Plasmodium falciparum (the parasite that causes malaria). I attribute their success to technical expertise...thoughtful single guide RNA (sgRNA) design, and the abnormally low GC content of the Plasmodium falciparum genome...Examples: “organism agnostic”, “model organisms”, “small genome”, “large genome”, and “organism biased scoring...Ghorbal, Mehdi, et al. "Genome editing in the human malaria parasite Plasmodium falciparum using the CRISPR-Cas9...

... to test if a type of antibodies from an Andean mammal called alpaca could neutralize the SARS-CoV-2 virus...In the example above, for example, we fostered a formal scientific collaboration between two research groups...transient partnerships where a researcher makes a small but crucial contribution to someone else’s project...find the right expert and the lack of credit for small contributions. We strive to eliminate these two ...

...to deliver Cas9 and your gRNA of interest into mammalian cells. This vector was the top requested plasmid...) V2.0 - This plasmid from the Zhang lab is a mammalian vector that expresses both SpCas9 and your gRNAs...scrambled shRNA is used as a negative control for mammalian RNAi experiments.  Find the scrambled shRNA vector...2A-GFP (PX461). pSpCas9(BB)-2A-Puro (PX459) - This mammalian expression vector encodes both SpCas9 and your...

...same cellular location, all at once! Small molecules What type of small molecules can you monitor with biosensors...thing that you care about within your cellular or animal system. This ‘thing’ could be a neurotransmitter... those which can be applied in cell culture and animal models primarily. Fun fact: Almost all of the biosensors...

...genetically engineered receptors to interact with small molecules to elicit a response in a certain cell...genetically modified receptors are activated by small molecules to either activate or inhibit neuronal...to respond to specific, pharmacologically-inert, small molecules. These new receptors were termed Designer...muscarinic receptor family and are activated by small molecule ligands such as clozapine-N-oxide (CNO)...