We narrowed to 1,073 results for: TOR;

...in epidermal growth factor receptor (EGFR) with the ClonTracer library and monitored population dynamics...paths to become resistant to epidermal growth factor receptor inhibition." Nature medicine (2016). PubMed...Library, deposited by Carrie Bhang, a research investigator in the In Vivo Pharmacology group at Novartis...mammalian cells through lentiviral infection and to monitor their abundance and clonal dynamics over time using...drug resistance, Bhang says “the [drug] resistance story panned out quickly,” and was summarized in a recent.... The library consists of a pool of lentiviral vectors with roughly 73 million semi-random, 30 bp DNA ... abundance of each barcoded clone can then be monitored over time by sequencing the barcodes in the population...

...recombination step, the effector is expressed. In cells where the stop cassette remains, the effector is not expressed...inactivate any read-through transcripts of the effector gene. Effector genes, any gene that you want to control...organisms by replacing the transcription factor binding site and effector in the construct. In Drosophila, the...T5 neurons by placing the calcium indicator GCaMP6f in the effector position, as well as to optogenetically...CsChrimson) attached to a fluorescent indicator (tdTomato), as the effector. They also expanded the SPARC system...A predictable number of cells will express the effector or retain the stop cassette following recombination...Drosophila (Fig. 1):  GAL4-UAS, a transcription factor binding site. GAL4 expressed in the fly binds to...

...G base editor (ABE) followed based on similar principles (Gaudelli et al., 2017). These editors did not... universities, private companies, and federal regulators. But the speed is impressive on a larger scale...2013; reviewed in Lander, 2016). The first base editor was reported three years later in 2016 (Komor et...access were all challenges for these early base editors. Further innovations were needed! Minimizing off-target...important addition. Improved activity of adenine base editors The first ABE was about 50% efficient in human ...mean better chances of identifying a successful editor. For this application, ABE8e turned out to be most...Targeting the right PAM Expanding PAM preferences Base editors can only access a short sequence, between four ...

...of pExpression vectors) plasmids. The type IIS restriction sites used in these vectors are of low abundance...of the many useful kits you can order from the repository: FX Cloning Kit BIOFAB Plasmid Set pCri System...to produce the constructs you need. Expression vectors within this kit contain a variety of promoters ...insects, and even Xenopus oocytes. The expression vectors additionally contain various combinations of protein...blocker isolated from a designed repeat protein combinatorial library by in vivo functional screen." Scientific...Series Confronted with the problem of creating vectors for predictable protein expression in bacteria,...this so-called bicistronic design (BCD). The 49 vectors in this kit can be used to drive production of ...

...genes with minimal effort. The versatility of Cas effectors allows for a wide variety of screens, including...packaging capacity and stable expression of lentiviral vectors make them an attractive option. However, safety... particularly in vivo ones. In those cases, AAV vector screens may be AAVantageous!       In vivo, in ...delivery method for in vivo CRISPR screens. AAV vectors can be made at high titers equal to those achieved... customization that is unmatched by other viral vector delivery methods. Lentivirus has limited packaging...packaging options, with most lentiviral vectors being packaged using VSV-G. In contrast, AAV can be packaged...psuedotypes to further improve the tropism of AAV vectors, enhancing their affinity for specific cell and...

...controlled by still other regulatory factors: enhancers, boundary elements, insulators, and silencers; however...tetracycline operators. Transcription can be turned on or off depending on what tet transactivator is used.... controlled by lac operator. Can be induced by IPTG. Commonly found in pET vectors. Very tightly regulated...the promoter (RNA polymerase binding site) and operators (response elements). Promoters are about 100 to...binding of the RNA polymerase and transcription factors, therefore promoters play a large role in determining...we will discuss these in a future post on viral vector parts. Promoter specificity Aside from choosing...Strong mammalian expression from human elongation factor 1 alpha Constitutive  Tends to give consistent ...

...I’ve learned the value in finding mentors wherever I go. This story by Dr. Erika Moore in Science Careers....351.6276.994 Moore E (2021) One mentor isn’t enough. Here’s how I built a network of mentors. Science. https://doi.org...undergraduate years. We each have a unique “origin story,” but taking the first step into the world of academic...lot of help from my peers, professors, and other mentors, and four years after I started college, I will...postdoc” is, and that “PI” stood for principal investigator. In basic science (as opposed to translational...in a lab. Position Common tasks Principle investigator (PI) Runs the lab Writes a lot of grants Varying...teaches graduate and/or undergraduate classes Postdoctoral researcher (postdoc) Getting more scientific...

...homology to the vector or neighboring fragment. Then you would amplify the fragments and vector by PCR, verify...linearized vector, the Cas9 enzyme along with a specific gRNA was used to cleave the 22kb vector. When followed...and then assembled into a vector using Gibson assembly. In this case the vector was PCR amplified to contain...be used in basic cloning of an insert into your vector of choice as shown in the figure below. To start...you want to insert your gene of interest into a vector with a large tag at the N-terminus, but you don...don’t have the tag already included in the vector you want to use. If you inserted these two pieces of ...gene of interest and the tag sequences into the vector in one step without scars as depicted below. First...

...plasmids are formed. Viral vectors are prone to recombination In lentiviral vectors, recombination often occurs...problems that I help troubleshoot involves viral vectors and plasmids with repeating sequences. Repeating.... Over time, bacteria containing the recombined vector backbone will also outgrow bacteria that contain...into two smaller plasmids. The ~3.7 kb recombined vector backbone contains the antibiotic resistance gene...samples? Plasmid recombination can affect viral vectors (LTRs and ITRs), plasmids with multiple polyA sites...recommendations from the depositing lab. If your vector is large (>10 kb) or has a low copy number, try...that in Stbl3 bacterial cultures with lentiviral vectors, the flat white colonies contained the full plasmid...

...initial reversal drive cannot restore the exact original sequence, it can restore the original protein-coding...are eliminated by natural selection, gene drives distort inheritance in their favor and consequently can...carrying a synthetic gene drive were to escape the laboratory, the drive could eventually spread through the...mistake would necessarily extend far beyond the laboratory and seriously damage public trust in scientists...gene drive is also the easiest: don't use a DNA vector that encodes both Cas9 and sgRNA. What if I want... can spread through populations of transgenic laboratory organisms but not wild organisms. For example...synthetic sequence will only spread in transgenic laboratory populations with that sequence2. Both methods...