We narrowed to 912 results for: IRE;

...choice. This step is sometimes also referred to as “directed differentiation.” Specific cell media, supplements...transdifferentiation adult somatic cells are converted directly into a different cell type, bypassing the lengthy... carrying the transdifferentiation factors are directly injected into the organ of interest for in vivo...Highly efficient reprogramming to pluripotency and directed differentiation of human cells with synthetic ....N. Davis-Dusenbery, and K.C. Eggan, SnapShot: directed differentiation of pluripotent stem cells. Cell...PubMed PMID: 19255591. 17. Spence, J.R., et al., Directed differentiation of human pluripotent stem cells...PMCID: PMC3192723. 21. Grigoriadis, A.E., et al., Directed differentiation of hematopoietic precursors and...

...is located directly upstream of the fluorescent protein genes, Cre recombination is required before fluorescence... (XFP) of different colors to label an organ or entire organism. Because each cell can have multiple copies...transgene that will recombine independently, cells may acquire one of a variety of colors when a combination of...expressed. Each cell remains the same color for its entire lifetime and daughter cells retain the same color...tandem with a single promoter, but only the gene directly following the promoter is expressed. Cre-mediated...downstream in the construct. As RFP and OFP are directly downstream of the promoter in Brainbow 1.0 and...expressed in a single cell maintains its color over the entire length of the cell, even axons of up to 100 microns...

... professors how they hire scientists for their labs. Tom Ellis selects new hires from a pool of CVs collected...three lab heads how they go about selecting new hires. This is followed by some perhaps surprising, yet...for both parties, he will invite the prospective hire to the lab to see how they fit in with the team....are dedicated and interested in science. “Postdoc hires are simpler because of their documented history ...with their interests.” George Church selects new hires initially based on those who notice and embrace ...technology development, etc). Second, the potential hire visits the lab for a full day, gives a 1 hour talk... typically offer more flexibility in choosing direction of a project. If you expect more 1:1 time with...

...genes. However, they are different viruses and require different isoforms of these packaging components... , pol , and env . The LTRs flank all of these required genes, and anything in between will be integrated...genome. To produce gamma-retroviral vectors, these required components have been separated into distinct plasmids...HEK293T Gamma-retroviral vectors can be packaged directly in human embryonic kidney 293T (HEK293T) cells...therefore, reduces the number of plasmids that are required in the transfection step of the viral packaging... gene therapy applications to more specifically direct delivery of gene therapies. Common Uses of Gamma-Retroviral...

...CRISPR/Cas9-introduced breaks are repaired. The following factors are required for NHEJ repair regardless of...pair of DNA ends together, forming a paired end complex The paired end complex then ligates compatible ...Non-Homologous End Joining (NHEJ) and Homology Directed Repair (HDR) – and how these pathways work, as...a higher capacity for repair, as there is no requirement for a repair template (sister chromatid or homologue...ends together. Similarly, it may be possible to direct insertion of an exogenous DNA fragment at a Cas9... structure of broken ends impacts how they are repaired. He works in Dale’s lab.        Dale Ramsden ...Blog Posts from Addgene: CRISPR 101: Homology Directed Repair CRISPR Protocol for Genomic Deletions...

...applications that require minimal cellular perturbation. Also, this method requires live, intact cells...attB-attP pair is aligned (sites facing the same direction) on a register, then the result of their recombination...attB-attP pair is anti-aligned (sites facing opposite directions) on a register, then the result of their recombination...sequencing is simple and the most accurate method as it directly reads the state of a register. However, each Sanger...difficult to isolate single registers or if it requires reading so many registers that Sanger sequencing...backbone) contains the register. Depending on the desired application, the inducible promoters we used to... be replaced by sensors that correspond to the desired inputs. Such sensors do not necessarily have to...

... allow for further purification, if desired, but it's not required for their use. Each RANbody is detectable...are a non-renewable reagent - Their production requires animal sacrifice and because they are an animal... for a summary. HRP HRP tagged RANbodies don’t require a secondary antibody for detection. Instead they...multi-labeling immunohistochemical protocols where either a directly labeled primary or a primary/labeled secondary...While Spaghetti Monster RANbody detection with a directly labeled anti-HA or -MYC antibody is less sensitive...synaptic partners (GRASP) assays No antibody required for detection RANbody is detected enzymatically...secondary Less sensitive than HRP-RANbody staining Requires secondary antibody staining HA Tagged GFP, RFP...

... for exclusively less-than comparisons. Paired and unpaired t-tests Samples that come from the same subject...You’ve acquired your long-awaited data – and it looks great! There are some exciting trends in your experimental...certain information about. The assumptions and requirements that need to be met for a parametric test are... - Quantitative If your data doesn’t fit the requirements above, a non-parametric test may be the way ...only care if one group is different in a certain direction (i.e., you just want to measure if a sample is...treatment, for example - should be analyzed with a paired t-test, because they eliminate subject-to-subject... on the other hand, should be analyzed with an unpaired t-test, which does account for subject-to-subject...

...component required (e.g. dsODN), doesn't require high read counts (fast multiplexing) Requires large cell...genome for all DSBs, doesn't require high read counts, fast multiplexing Requires delivery of dsODN (potentially...length of the gRNA given the stricter sequence requirement at the terminal 3’ PAM site (Fu et al. 2013; ...by sequencing (GUIDE-seq) (Tsai et al. 2015) and direct in situ breaks labeling, enrichment on streptavidin...BLESS detect double-stranded breaks and do not require high sequencing read counts making them fast and...contains both the rare off target event and the desired edit is low. Therefore, unbiased sequencing may...selected. Conversely, translational research may require the rigor of both forms of off-target analysis ...

...Klompe et al., 2019; Strecker et al., 2019), which direct RNA-guided transposition. By natively combining...I-F CRISPR-Cas system. The four major components required for RNA-guided DNA integration are (Fig. 1A, top...transposition in type I-F INTEGRATE. The requirement for multiple Cas proteins to form the Cascade...targeting (Halpin-Healy et al., 2020). Genomic targets require an upstream 2-bp PAM sequence for recognition by...instead of three Cas proteins (Fig. 1A, bottom), and require only two transposase proteins (TnsB and C). While...effector plasmid (pEffector) expresses all seven required genes and the CRISPR RNA under the same promoter...: Clone spacer(s) into the CRISPR array Clone desired DNA cargo into the mini-transposon Transform plasmid...

...editing systems; first, RNA editing doesn't require homology-directed repair (HDR) machinery, and could thus...in non-dividing cells. Cas13 enzymes also don’t require a PAM sequence at the target locus, making them... sequence, but orthologs like LwaCas13a do not require a specific PFS. Cas13 enzymes do not contain the...domains responsible for DNA cleavage, so they cannot directly edit the genome. A Cas13-based RNA editing system...enzyme for further studies. PspCas13b does not require a PFS, and it is sensitive to mismatches in target...the lowest amount of off-target editing. In one direct transcriptome-wide comparison, REPAIRv2 reduced...activity. They then performed sixteen rounds of directed evolution across the full ADAR2DD to further optimize...

...fluorescent donor to your protein is to hardwire the molecule directly into the protein’s peptide sequence...(the donor) is excited, it can transfer energy directly via resonance to the second fluorophore (the acceptor...translation (Ermolenko et al., 2007). All that is required is that the proteins/nucleic acids/lipids of interest...delivered to the right place? FRET experiments require targeting the donor and acceptor fluorophores to...experimental design is that the targeting specificity requirements are different for the donor and acceptor fluorophores...codon (often amber i.e. UAG) engineered into a desired site. During translation, the novel tRNA/unnatural...overhaul of the cell’s native tRNA synthetases is required and thus far, this method has proven most effective...

... ori and requires the host proteins DnaA and IHF to initiate replication. But pSC101 requires a third ...RepA binds to directly repeated sequences called iterons located near the ori and directly interacts with...said to be under relaxed control, as they only require the host's replication machinery for elongation...have enough nutrients available to complete the entire round of replication. Initiation is the primary...versions, but binding of the low-affinity versions is required to start the replication process. When enough ...creates two replication forks that move in opposite directions away from the ori. In the elongation phase, a...on host-encoded proteins, it can be regulated indirectly via the same mechanisms that affect DnaA and ...

...is necessary to be direct and clear. There is place in human discourse for indirect language, but it is...Giving timely and useful feedback is an absolutely required aspect of being a successful manager. Great bosses...over, what would you do differently? Beware of indirect language You must be honest about performance ... should be aware that they are at risk of being fired and should be looking for a new position. One of... mistakes I see is managers who hint at or use indirect language to talk about real performance issues...issues. Watch this little video to learn more about indirect language from Steven Pinker. As a manager you ...Scientists. Subscribe to the Addgene Blog to follow the entire series as it is published. For more reading try...

...practices, their opinions towards ELNs, and their desired functionalities i.e. main priorities of different..., and the fact that long-term use is likely to require on-going maintenance and support.” Besides that...it as very important. One comment reflected the desire for a flexible generic solution, rather than an...switching from paper to an electronic lab notebook requires time and knowledge. In a recent article, we translated...reporting on a project – printing your data out can be required every once in a while, so it is good to make sure...your lab need to meet any specific compliance requirements – e.g. 21 CFR part 11, HIPAA? Answering this... that take any ownership over your data or have direct access to it. Your data should be kept private ...

... for a site-directed mutagenesis PCR that uses mutagenic primers to introduce all desired codon changes...generation of a plasmid library that expresses all desired variants of a protein. Applying a selective pressure...phage-derived uracil labeled template, this method requires further DNA shuffling to combine pairs of mutations...of your liking. This method is similar to site-directed mutagenesis (SDM) PCR, but with a twist since ...the plasmid. Again, if they are in the opposite direction, both strands of the plasmid will be nicked during...of your plasmid. If the sites are in opposite directions, you just need to use a plasmid with BbvCI restriction... the gene or use custom gene synthesis or site directed mutagenesis to remove the BbvCI site(s). Overall...

...but this approach would require robust stem cell editing to maintain the desired phenotype. Precision genome...skipping. Addgene depositor Charles Gersbach used paired zinc finger nucleases to remove exon 51 in DMD ...For such a therapy to be successful, multiple requirements must be met. First, CRISPR would need to be ...especially well suited for genome editing. The homology-directed repair (HDR) pathway is downregulated in mature...is maintained for at least six months. Future directions and obstacles to clinical CRISPR editing Given...NHEJ-mediated editing, but most other diseases will require HDR-mediated precision editing tailored to smaller...:10.1126/science.aae0169 2. Long C, Amoasii L, Mireault AA, McAnally JR, Li H, Sanchez-Ortiz E, Bhattacharyya...

...the requirement for digestive motility to not be disturbed, the brain fibers could not be directly repurposed...constantly influenced by signals it receives from the entire body, making it difficult to probe due to the lack...monitor multiple systems. Ideally this tool would be wireless, able to monitor outcomes of interest, and stimulate...blocks of polymers are heated up and pulled to a desired thickness. The fibers are drawn with microelectronics...the fiber horizon? The brain interfaces with the entire body, so the obvious next applications are to expand.... P. (2020). Modular Optoelectronic System for Wireless, Programmable Neuromodulation During Free Behavior... Multifunctional microelectronic fibers enable wireless modulation of gut and brain neural circuits. Nature...

...appropriate plasmids, and code the entire page. That would be entirely too much work for one person, it ...Clearly articulate the desired outcomes. Begin with the end in mind. Specify desired results. Are you delegating...and are experts in each of the individual tasks required to develop the web page. The team manager must... expert) the end result is high quality and the entire process is completed more efficiently. Listen to...criteria for delegating tasks appropriately: Are the desired goals and outcomes clear? If you don’t have clear...results. What does success look like? Agree on desired outcomes. Concern yourself with what is accomplished...

...absolute measure of target abundance, does not require standard curves, and can theoretically detect a...what kind of assays can be used to sniff out your desired mutations. The assays you will use aren’t that ...either a donor sequence integrated through homology-directed repair (HDR) (1), or indel mutations resulting...non-homologous end joining (NHEJ) (2-5), depending on your desired genome edit. Since exploitation of NHEJ to generate... and a NHEJ/ “drop-off” probe designed to bind directly at the predicted double strand break site (Figure...quantification. Comparatively, mismatch assays require between 200 and 500 ng of purified PCR product.... those with the bi-allelic mutations typically desired when screening clones for complete functional knockout...