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...human therapeutics (e.g. using Sleeping Beauty). Despite their diverse utility, DNA transposons have been...(V. cholerae INTEGRATE - hereafter VchINT) and encodes a Type I-F CRISPR-Cas system. The four major components...the same promoter, and a donor plasmid (pDonor) encodes the mini-transposon. pEffector and pDonor can also...: Clone spacer(s) into the CRISPR array Clone desired DNA cargo into the mini-transposon Transform plasmid...even very large genetic cargo, at one or multiple desired genomic loci. Many applications of these systems...

...individual molecules. Poor photon budgets are a widespread issue with fluorescent proteins—even the thriftiest...student searching couch cushions for spare change, desperate to extract a few more photons from a sample. Chemical... the complexity of the system. A particularly desirable property for small-molecule labeling strategies...size, brightness, and cell permeability. In past decades, further refinements to this established dye structure...spectral range. Nevertheless, these fluorophores were designed as antibody and oligonucleotide labels for fixed-cell...

...beta-lactam resistant population to the antibiotic, despite this resistance gene being encoded by a high-copy...currently undergoing a renaissance. Yosef et al. designed a two-phage CRISPR system to resensitize antibiotic-resistant...immune system, may someday be used to target the hardest-to-kill bacteria. In using phage-derived vectors...Ahmed A. Gomaa, and Rodolphe Barrangou. “A CRISPR design for next-generation antimicrobials.” Genome Biol...Research Read Our CRISPR Guide Pages Find CRISPR gRNA Design Tools ...

...DNA bases without breaking the DNA backbone, sidestepping the problem of HDR vs. NHEJ entirely. The first...editors with editing windows as small as 1-2 nucleotides. To reduce off-target effects associated with...fourth-generation base editors, BE4, reduce the undesired C→G or C→A conversions that can happen with BE3s...named ABEmax. In 2020, two papers were published describing additional ABEs evolved from ABE7.10 that have...cytosine and adenine base editing, called CABE-Ts, TadDEs, or TadCABEs, without additional fused components...editors have a narrow editing window of only a few nucleotides, so they are tightly constrained by PAM availability...

...wealth of information and includes annotated images of the brain sections describing the labeled regions, ...the human, mouse, and macaque genomes, and are designated by a letter following the enhancer ID (h, m, ...enhancers and their neuroscience research. This includes other data, tools, and resources. Luckily for ...no doubt changed drastically over the past few decades. But no matter how much it changes, it will still...

...manner. These videos can be especially useful when designing new primers to clone your sequence of interest... your cloning. Most sequence analysis software includes Golden Gate cloning tools that simulate the restriction–ligation...according to their concentration, size, and the desired insert-to-vector ratio using the equation in Figure..., positive colonies may contain plasmids with undesired ligations, so always screen multiple colonies ...see in colony PCR electrophoresis), is free of undesired mutations, and that you have the correct four-...

...1: CROPseq-multi uses two sgRNAs with internal barcodes (iBARs), multiplexed using tRNAs, within the lentiviral...separation of the multiplexed elements. The team designed sgRNAs that include an internal barcode and separated... have been made since prime editing was first described in Anzalone et al., 2019, including modifications... sparsely-labeled field of view that was more desirable than having many closely-packed labeled cells.... combination with miR124T targeting sequences, provides a simple yet valuable tool for microglia research...

...Engineering Solutions, which also includes our CRISPR knockout guide design tool, guide validation tool, and...their relative abundances.   The ICE tool also provides a Knockout score (KO-score) which represents the...the CRISPR workflow. For a multiplex sample, ICE includes visual representations of all detected edit types...Contributions, Indel Distribution and Traces - provides particular details about the indel profile of ...

...second outcome and increases the generation of the desired T-A base pair. Figure 1: Transition mutation...by base editors. Pathways that compete with the desired outcome are not pictured. Adenine base editors... SpCas9-VRER TadA-TadA* NGCG 8  Tips for designing base editing experiments in plants Choose a base...successful base editing causes the creation or destruction of any restriction enzyme (RE) recognition site... indel sometimes may also results in RE site destruction. Many thanks to our guest blogger Kutubuddin...

...Over the past decades, neuroscientists have made great strides to map brain regions and allocate specific...for this task. However, most lesioning methods destroy areas larger than intended and may unintentionally...surface. The latter reflects a state of behavioral despair, and the more time the animal spends in the passive...that were not (Hare et al., 2019). The research described here is only a small selection of the increasing...

... defined as microbial survival despite exposure to antibiotics designed to kill them or to impede their...that the term ‘antibiotic’ can also be used to describe anti-parasitic drugs and anti-fungal drugs, because...different reasons. Exposure to antibacterial drugs provides a selection pressure, killing off bacteria that...equipment, etc. Virtually any preventive measure designed to help keep you from getting sick can be helpful...

...They also host AMA forums where users can ask designated scientists anything about a particular topic....calendar invite over to the r/Science team with a description of our AMA and Joanne’s preferred time to start...was writing all day and straight on through her designated two hours for answering questions. She was able...toxic advisors, fruitless job searches, and even despair at having chosen an academic past. I hope I was...receive via email. The cool features are too many to describe here, but I have over 2,000 notes which Evernote...

...data from your FRET experiments. You may have to design several different constructs to find one that could...studies. These parameters are less important when designing a biosensor, as the distance between the two fluorophores...gene of interest: Plasmid Color Expression Description pPROEX Aqua Cyan Bacterial Expresses Aquamarine... straightforward and quantitative, but it is destructive and cannot be used for dynamic measurements. .... Extra care should be taken so as not to destroy the donor molecule. Fluorescence lifetime imaging microscopy...

...insert into RNA. In practice, the term "promoter" describes the combination of the promoter (RNA polymerase...   Primarily used for RNA     transcript     Description Expression Additional considerations CMV General... phosphoglycerate kinase gene.  Constitutive  Widespread expression, but may vary by cell type. Tends ...promoters     Promoter     Primarily used for Description Expression Additional considerations T7 in ...identify the right tissue-specific promoters as described by researchers at the NIH here.  Note: A. Max...

...when it comes to fluorophore selection for panel design. Find streptavidin plasmids and biotin plasmids... this system is increased flexibility for panel design. Another perk is that one primary antibody can ...for example, a previously induced gene knockout. Besides amine- and DNA-binding dyes, nucleoside analogs...walk through the topics we covered in this post. Besides direct labeling, indirect labeling can be used ...surface, dump gating is a useful way to get rid of undesired cell populations. Dump gating uses the same color...

...recombinant antibody. They are ~25 kDa single polypeptides that contain the variable light chain (VL) and...lethal to animals.  In the clinic, scFvs’ size also provides advantages over antibodies (Ahmed et al., 2012... bond between the VL and VH domains. This bond provides stability and solubility for the scFv (Gaciarz...clinic (Bates and Power., 2019).Many of the scFvs designed for oncology applications are reformatted as bispecific...Chantima W, Sangma C, Poomputsa K, Dharakul T (2015) Design and Generation of Humanized Single-chain Fv Derived...

...required, through mitochondrial transfer. In the two decades since this transfer was first proposed, researchers..., 2025). These new sensors use quite different designs, leading to unique advantages. While most red GECIs...two-photon microscopy, and fiber photometry. Plus, both designs offer promising templates for future improvements...al. (2025) under CC BY license. The paper includes other great feats of materials engineering as ... relative simplicity as a gene editing system. Despite that simplicity, its accessibility outside of well-equipped...

...protein analyses using ELISA. Peptides 72:4–15 . https://doi.org/10.1016/j.peptides.2015.04.012 Burnette WN ... Figure 2: The western blot dry apparatus includes a stack of filter paper, a copper anode/iblot ...blocking buffers, and incubation times. If the vendor includes publications that cited the antibody, consider...abundance In addition, the thickness of the band provides information about the relative abundance of the...

...identify possible gene function. In this post, I will describe a useful and cost-effective method that our lab...and probe with live cells The following is a description of a technique published by Simpson, Sacher, ...ends) and ligate into a bluntly digested vector designed for protein overexpression in E. coli, such as...the original plate to identify which phage gene encodes that protein! First, block your membranes with ... can lead to high background that obscures the desired signal. We have successfully corrected this by ...

... the work, shared the credit, and we both won accolades that would launch our careers in science. Fast...helped assemble a crack team of Addgenies that includes: Michelle, the Senior Scientist who coordinates... a happier person. What does a good team need? Besides that all-important sense of purpose, there are ...Slack constantly. Whether your teammates are two desks away or on the other side of the world, communication...self-interest ahead of the team’s goals, then that team is destined to do poorly. To some extent, it doesn’t even ...