...form or as ready-to-use lentivirus. Figure 2: dCas9-Tet1 demethylates cytosines at promoters and...pX330a dCas9-LSD1 for targeted removal of H3K4me1/2 and H3K9me2 histone modifications. Like the TALE-LSD1...approach for targeted DNA demethylation. Cell Discovery 2: . https://doi.org/10.1038/celldisc.2016.9 Find plasmids... and targeting the epigenome may make certain processes, like cellular reprogramming of iPSCs, more efficient...

...certain states of interest. For example, in Figure 2 below we present a two-input, five-state GRSM register...linking RSM inputs to endogenous gene expression (2). Our paper (1) explains how to arrange attB-attP...Science. 353, aad8559 (2016). PubMed PMID: 27463678. 2. Green, Alexander A., et al. "Toehold switches: de-novo-designed...used to better understand and engineer cellular processes that are influenced by temporally ordered biochemical...

...scientists have 1) the courage to make mistakes and 2) the integrity to recognize these mistakes quickly...She studies beta-cell function and failure in Type 2 Diabetes. Birmingham, AL, USA 33.5207°N 86.8025°W ...available to people who want to join the dialog and 2) remember that science should not be partisan: everyone...might feel disconnected from the entire scientific process.  I've spent a lot of time debating how to fix ... science in May, I marched to advocate for the process we use to learn about the world. In a way, I marched...

...’re delivering their payload to.   Figure 2: Numbers of AAV tools, by category, available from... substrates, light, or engineered ligands (Figure 2). Biosensors, which include calcium and voltage sensors...selection process to standardize our feedback and evaluation of viral vectors. This process builds on ...significant time and removes a variable from the lengthy process of validating and using an AAV in a specific experimental...its inherent bias: Addgene uses a prioritization process to decide which tools we make available. This ...us first identifying broadly useful vectors, a process that isn’t as simple as it seems. Take, for instance... and blog posts. The lessons learned from this process have shaped and supported our growth and development...

...the chromosome, ending replication.   Figure 2: Prokaryotic chromosomal replication. Adapted from...Microbiology and Molecular Biology Reviews: MMBR, 62(2), 434–464. https://doi.org/10.1128/MMBR.62.2.434-464.1998...control point of DNA replication in bacteria, so the process is carefully regulated by a mix of positive and...replication (oriC), where initiation begins. This process relies on the initiation protein DnaA. First, DnaA... versions is required to start the replication process. When enough copies of ATP-DnaA are bound, including...integration host factor (IHF) assists with the process: it bends the DNA to redistribute the DnaA from...plasmid ori, after which replication of the plasmid proceeds (and terminates) similarly to the bacterial chromosome...

...residues, which can be found throughout the antibody. 2) Endogenous cysteines, such as those that make up ...Detecting antigens by quantitative immuno-PCR. Nat Protoc 2:1918–1930. https://doi.org/10.1038/nprot.2007.267 ...yourself, there are kits available to make the process quick and easy. Before getting started, be sure...

...studies have shown that the conditions Sotos-type 2 and Marshall Smith Syndrome are due to mutations in...accurate diagnosis of NFIX-related syndromes (Sotos-type 2 and Marshall-Smith) and could be expanded to NFIA-...lines and use these lines to study how immune processes and biological pathways are altered in PMM2-CDG...

... that all look exactly alike.      Figure 2: Reagent gradient. Here, the lanes are lightly marked...guide. The gel is then removed from the membrane (2) and the membrane is cut so each lane is its own piece...in the bands, you may want to consult Tables 1 and 2 in our Technical Design of a Western Blot post to ...the antibody’s dynamic range. Specifically, the 1:2, 1:1, and No dilution samples all have approximately...conditions are 4 °C overnight or room temperature for 1–2 hours, there is some evidence that antibodies may ...high background or low signal during the imaging process, try adjusting your exposure time first, since ...

...antibiotic selection. Essentially, this gene is a 2-in-1 providing both bacterial and eukaryotic selection...Zeocin-induced DNA cleavage in human cells. Mutagenesis, 20(2), 111–114. https://doi.org/10.1093/mutage/gei016I .... During the propagation and/or transformation process, some bacteria will either never receive a copy...antibiotics listed below affect continuous cellular processes (e.g., protein synthesis), while amp only affects...

...with neuron-specific expression of dCas9-VPR (Figure 2), a popular Cas9 activator developed by the Church...ubiquitous ones such as EF-1α and CAG. Figure 2: Dual lentivirus system with ubiquitous expression..., studying the genetic underpinnings of these processes has been complicated by the lack of precise genetic...undifferentiated cells at the end of the screening process. However, while the neuronal marker TUJ1 is useful...

...green squares, purple circles and blue triangles. 2) An antibody immobilized on a bead binds specifically...protein G, a novel IgG-binding reagent. J Immunol 133(2):969-974. Hjelm H, Hjelm K, Sjöquist J (1972) Protein...to immunoprecipitation and simply refers to the process of anchoring an antibody, often called the capture...protein interactions, and limit high throughput procedures. However, agarose beads are often a more cost-effective...

... reduce the background staining (Table 2).      Table 2: Secondary antibody selections for a four-antibody...have been cross-adsorbed. The cross-adsorption process filters out off-target antibodies increasing specificity...

...wheat germ, HeLa, and Leishmania (Gagoski, et al 2015[2]). The proteins encoded in this set range in size ...to generate Illumina sequencing libraries using a 2-step PCR-based protocol and are compatible with single-end...decrease false positives observed in the cloning process, making it easier to produce the constructs you...novel and powerful pyranopyridine derivatives." Proceedings of the National Academy of Sciences 113.13 (2016...-time observation of metabolite production." Proceedings of the National Academy of Sciences 113.9 (2016...pCri system to simplify the protein purification process by making it easy for a researcher to quickly generate...on the system Easing the Protein Purification Process with pCri Cell-free protein expression kit Cell-free...

...break), increasing editing efficiencies 2-3-fold. However, the process of double nicking increases indel formation... editing as efficient as possible. Figure 2: Decision tree for selecting a PE6 variant. PE = prime...using paired prime editing. Nature Biotechnology, 40(2), 218–226. https://doi.org/10.1038/s41587-021-01025...the template and primer binding site, increase processivity, or improve thermostability. What came out on...repair (MMR) complex, to temporarily inhibit this process and promote copying of the edited strand to the...to the numerous innovations that streamline the process of prime editing, the Liu lab contributed an improved...

...whether the results include knockout data.    Figure 2: Search options in the Antibody Data Hub. Data reports...antibody worked in your application. The submission process is easy, with simple, descriptive fields, drop-..., and can be saved at any point throughout the process, so you can work through it at your own pace. To...

...gene activation, with some genes experiencing around 2-fold activation levels. Although other methods have.... This method has been shown to activate genes at 2- to 3-fold higher than SAM, SunTag, and VPR (Zhou ...activation across multiple loci was an arduous process. When using zinc finger proteins or TALE proteins...

... digest and compare it to some reliable controls. 2. Viral stocks may be sensitive to freeze thaw cycles...PMID: 24003179. PubMed Central PMCID: PMC3760253. 2. Rohwer, Forest. "Global phage diversity." Cell 113.2...volume. Always remove packaging cell debris before proceeding to ultracentrifugation to avoid contaminating...

...estimate transformation efficiency (Figure 2).    Figure 2: Example of a serial dilution to estimate... reduce recombination during the amplification process. In liquid culture, all of the bacteria can grow...

... PITCh vector via In-Fusion or SLIC cloning. Step 2: Design an insertion locus-specific gRNA The gRNA ...Find the plasmids from this publication at Addgene. 2. Dalvai, Mathieu, et al. (2015). “A Scalable Genome-Editing...MMEJ doesn’t get a lot of press. However, this process accounts for a percentage of the mutations seen...

...from GFP and the mutation definitions found in Table 2 may not apply. Table 1: The specific mutations comprising..., V163A EBFP F64L, S65T, Y66H, Y145F   Table 2: Functional role of specific mutations in GFP derivatives... enabling the study of dynamic, physiological processes. Re-engineering GFP to increase its range of color...