We narrowed to 580 results for: vit;
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TypeBlog Post... protein activity and have important roles during oxidative stress. To monitor this activity, Pedre Pérez...biosensors that detect new redox metabolites are vital for researchers to better understand the processes...assessed cFos expression. She will explore the connectivity patterns of these cells and the functional contribution... will allow us to better understand how neuron activity changes following opioid withdrawal, providing...excitatory and inhibitory neurons whose function and connectivity are poorly understood,” says Qiu. Qiu developed...both optogenetics and chemogenetics to study the activity of parvalbumin (PV)-expressing neurons, neurons...
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Enhancing CRISPR Targeting Specificity with eSpCas9, SpCas9-HF1, & HypaCas9
TypeBlog Post...retained the on-target activity of wt Cas9 while decreasing off-target activity at known off-target loci...displays comparable on-target activity and similar or decreased off-target activity across the sites tested ...effects of CRISPR/Cas9 by altering the nuclease activity of the Cas9 nuclease itself. Design and testing...Created with BioRender.com. To decrease the electropositivity of the HNH/RuvC groove, Slaymaker et al. made...combination mutants for their on-target and off-target activity at multiple genomic locations in HEK cells. Further... -
Hot Plasmids: Winter 2025
TypeBlog Post...and responsive voltage indicator with enhanced sensitivity for unitary synaptic events By Brian O’Neill ...ASAP5 selected for its improved kinetics and responsivity in screening ASAP3 variants. B) Electrical and... and optical recordings of spontaneous activity in human stem cell-derived neurons expressing ASAP5-Kv...regimes with fast activation kinetics and high responsivity near the resting membrane potential, ASAP5 is...and responsive voltage indicator with enhanced sensitivity for unitary synaptic events. Neuron, 112(22),...context-dependent responses and molecular patterns of selectivity. With 10 plasmids available as a kit, plus 21...Direct interrogation of context-dependent GPCR activity with a universal biosensor platform. Cell, 187... -
New Tools Enable CRISPRa for Neuroscience Applications
TypeBlog Post...gRNAs in vivo, they should be screened for activity in vitro and then packaged into AAV for in vivo delivery...SPH mice CRISPRa has largely been limited to in vitro settings due to the large size of Cas9 activators...developed neuron-optimized lentiviral vectors for in vitro and in vivo CRISPRa (Savell et al., 2019). This ...Bdnf is expressed early in response to neuronal activity (hence, “immediate-early”) and consequently regulates...immediate-early genes associated with neuronal activity, such as Fos, Egr3, and Arc. Thus, CRISPRa represents...preferable for individual gene activation in select in vitro and in vivo applications, the tools described above... -
Viral Vector Guides and Plasmids
TypeCollection...and envelope plasmids). When transfected into in vitro cell cultures, these plasmids direct the production... -
Hot Plasmids and Viral Preps - July 2021
TypeBlog Post...biosensor has a high sensitivity (50 nM-500nM range) and a high specificity for SLs in vitro, and they further...SpCas9 variants have improved activity on NAG PAMS and reduced activity on NGG PAMs relative to wild type...mimic aspects of organ morphology and physiology in vitro and have emerged as a powerful model for understanding... the enzymatic biosensor retains its catalytic activity. As SLs are also known to trigger seed germination... -
mTOR Pathway
TypeCollection...tumor suppressor p53 also promotes aberrant mTORC1 activity, and multiple familial cancers are caused by inactivating... -
CRISPR Plasmids - Prime Edit
TypeCollection... and two mutations in Cas9 to enhance nuclease activity; can be used in PE2–PE5 systems PE6 - small prime... -
When Fidelity Matters: A frank discussion about ligase fidelity
TypeBlog Post...phosphorylated DNA termini to 3 ́-OH DNA termini (1-4). In vitro, ligases (notably T4 DNA ligase) are critical reagents...circularization of dsDNA (6). Less commonly utilized in vitro, Taq DNA ligase will ligate only nicks (5-8). Taq... applications, there cannot be any end-joining activity, and for others, there is a need for the exclusive...or qPCR-based methods to greatly increase the sensitivity of detection (14-20). LDR has also been extended...achieved through a combination of the intrinsic selectivity of the ligase active site and careful balance... that extends earlier studies through a high-sensitivity multiplexed format (21, 27). This methodology...Bruce Wallace. "Specificity of the nick-closing activity of bacteriophage T4 DNA ligase." Gene 76.2 (1989... -
Sequencing Options for CRISPR Genotyping
TypeBlog Post...Provided 5% Inexpensive, simple Low-throughput, low sensitivity Sanger Sequencing Provided 20% (variable) Sequence-level...inappropriate for heterogenous populations, low sensitivity Targeted Amplicon Sequencing Provided 0.01%...including Digenome-seq (Kim et al. 2015) for in vitro analysis, IDLV for in vivo detection (Gabriel et...potentially toxic) Digenome Seq 0.1% Cell-free (in vitro) Works across all cell types Must be verified with... background BLESS Not reported Cell-based (in vitro) Can be used on tissue from whole animal models,...throughout the genome. However, their decreased sensitivity means that the best option moving forward may...most investigators use single cell clones for in vitro CRISPR experiments. The likelihood that a single... -
25 Work From Home Tips for Scientists
TypeBlog Post...outdoors. Taking a walk or a bike ride is a great activity for both exercise and clearing your mind. And ...And because the outdoors is so huge, these are activities that you can easily do while staying away from... your life. At Addgene, we’ve started virtual activities like team lunches, a virtual soup lunch, and ...kids' needs. Plan ahead. Think about and set up activities that kids can do more independently. For example...meals and snacks ahead of time. Find learning activities for kids. For example, Skype a Scientist is now...research community. Set expectations for your productivity Set a routine and stick to it. Plan defined... -
15 Hot Plasmids from 2017
TypeBlog Post...off-target activity than BE3 85172 pBK-EQR-BE3 EQR-Cas9 (NGAG) Lower off-target activity than BE3 ...donors in experiments simultaneously measuring the activities of CaMKII and RhoA during induction of structural...domain similarly allows for control of enzymatic activity. In addition, proteins can be “caged” by fusion...editing. For each base editor, they observed editing activity with a minimum efficiency of ~50% and confirmed...85177 pBK-YEE-BE3 SpCas9 (NGG) Lower on-target activity than BE3; editing window ~1-2 nt 87438 pET42b-HF-BE3...pET42b-HF-BE3 HF-Cas9 (NGG) Very low off-target activity; protein purification vector 87439 pCMV-HF-BE3...pCMV-HF-BE3 HF-Cas9 (NGG) Very low off-target activity; mammalian expression vector Rees H, et al. Nature... -
Using AAV for Neuronal Tracing
TypeBlog Post...current understanding of neuroanatomy and neural connectivity, providing valuable insights for neuronal communication...methods have been developed to study neuronal connectivity. These methods are less neurotoxic and better...). When using these viruses to mark neuronal connectivity a large majority of neuronal cells are labeled...improvement resulted in a clearer picture of neuronal connectivity in the mammalian brain (Wickersham et al., 2007...tracing with AAV vectors is the Allen Mouse Brain Connectivity Atlas, a project to map neuronal connections... study can be found on the Allen Mouse Brain Connectivity Atlas website. A similar approach was used by...connections that are functionally relevant, i.e. activity-dependent neuronal tracing. Apart from mapping... -
Expanding the Targeting Scope and Editing Efficiency of Adenine Base Editors
TypeBlog Post...links the expression of gene III to base editor activity and editing speed and consists of the following...Figure 2: Phage-assisted evolution of base-editing activity. A plasmid-based genetic circuit in the bacteria...links the expression of gene III to base editor activity and editing speed. Created with BioRender.com....of TadA variants with much higher base editing activity compared to the original TadA-7.10 enzyme. After...showed increased off-target DNA and off-target RNA activity. To mitigate this drawback, the lab introduced...editor with improved Cas domain compatibility and activity. Nature Biotechnology. https://doi.org/10.1038... -
Promega Plasmid Collection
TypeCollection... measure subtle expression changes with high sensitivity. These assays are ideal for real-time monitoring... -
AAV Packaged on Request
TypeCollection...is used in a wide range of experiments, both in vitro and in vivo. It has low immunogenicity, is easy ... -
Hot Plasmids and Viral Preps - January 2021
TypeBlog Post...recombinase activity were exposed to blue light. Cre-Vvd showed substantially improved activity as compared...permitting recombinase activity. Opto-Cre-Vvd is precise, demonstrating low sensitivity to ambient light, ...gene expression tools in and of themselves, their activity can be fine-tuned and precisely controlled through...authors demonstrate that CRISPR-Casɸ is active in vitro as well as in human and plant cells. Casɸ... -
A Needle in a Base-Stack: Cas9 Structural Biology
TypeBlog Post...for in vitro cleavage, but they improve cutting efficiency and are important for robust activity in cells... helps limit off-target activity. What next? It’s not actually clear! In vitro, Cas9 holds tightly to ...cells (Jiang & Doudna, 2017). Within an interior cavity, Cas9 grips nucleotides 1-10 of the guide RNA, ...the HNH domain also allosterically controls RuvC activity through a hinge region between the two (Jiang ...finish: Figure 9: Cartoon summary of Cas9 activity. The apo enzyme is shown in the open conformation... -
The AAVantages of AAV in CRISPR Screens
TypeBlog Post... Overview of in vitro AAV CRISPR screens. Created with BioRender.com. The in vitro option requires... screens may be AAVantageous! In vivo, in vitro, ex vivo CRISPR screens can be carried out using...using a few different methods — in vivo, in vitro, and ex vivo. The general process of these screens is the...with transducing cells with the CRISPR library in vitro before transplanting them into mice. For in vivo...having an efficient and effective delivery method is vital for the best outcomes. The go-to in vivo option ... -
Viral Vectors 101: Voltage Indicators
TypeBlog Post...voltage sensitivity of the available VSDs has been successful even at the single neuron level in vitro. In...function depend on potential energy. Cellular activities like action potentials, muscle contractions, ...effective at the single cell level as they are in vitro. Factors like photobleaching, lack of red-shifted...genetically encoded component is delivered in vivo or in vitro and a small molecule is added exogenously to measure...red fluorescent voltage sensor to detect neural activity. Sci Rep 9, 15878 (2019). doi: 10.1038/s41598-...