...the approaches that can be used to determine selectivity of an antibody for flow cytometry experiments...well as a pragmatic approach to determining the selectivity of antibodies for other sample types and for ...For these reasons, it is essential to test the selectivity of your antibody. But testing is not one size...your experiment. An antibody that demonstrates selectivity for cell surface labeling may become less selective...knockdown to enable robust assessment of antibody selectivity. Additionally, it can be a real challenge to ...protein level at the timepoint studied Off-target activities of the RNAi Correctly interpreting your data...protein expression makes confirmation of antibody selectivity even more challenging. Correlation with RNA or...

...balance maximizing on-target activity while minimizing off-target activity, which sounds obvious but can...100nt window downstream of the TSS gives the most activity. Thus, a given gene will only have a dozen or ...the method of producing the guide (synthetic, in vitro transcription, or lentiviral delivery) can affect...unwise to make conclusions on the basis of the activity of a single gRNA, and thus diversity of gRNAs ...approaches. Avoiding off-target effects The off-target activity of gRNAs is important to consider. While the basic...mismatches that can nevertheless still lead to activity has been established, and can be used to identify... indicates that the consequences of off-target activity, at least for the experimental conditions used...

... receptor) NR3C2 Plasmids NR3C2, MCR, MLR, MR, NR3C2VIT nuclear receptor subfamily 3, group C, member ...

...CasPEDIA feature. Activity features: CasID CasPEDIA will display both the cis and trans activity of each entry...enzyme: Nuclease properties (CasID): Cis/trans activity, PAM requirement, gRNA, and multiplex ability ...strands of its dsDNA target in cis and has no trans activity. AsCas12a, on the other hand, generates staggered...manually enter details to refine your search (trans activity, multiplex ability, gRNA design, etc.). With this... an evolving database, and all researchers are invited to contribute to it! The CasPEDIA Consortium will...update entries as research progresses, but you are invited to reach out to suggest new entries or even new...

...partner labs. Learn More ID Recombinant Antibody Reactivity Recommended Applications PI Don’t See What You...

... 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...

...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...

...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...

...tumor suppressor p53 also promotes aberrant mTORC1 activity, and multiple familial cancers are caused by inactivating...

... and two mutations in Cas9 to enhance nuclease activity; can be used in PE2–PE5 systems PE6 - small prime...

...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...

...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...

...is used in a wide range of experiments, both in vitro and in vivo. It has low immunogenicity, is easy ...

...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...

...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...

...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...

...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...

...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...

...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...

...RNAs regardless of homology to the crRNA. This activity provides a stark contrast to Cas9 and Cpf1, which...