We narrowed to 814 results for: Ran;

...which cleaves one strand of DNA upon binding the protospacer. The resulting double strand break (DSB) stimulates...two catalytic domains, HNH and RuvC, cleave DNA strands complementary and non-complementary to the gRNA... binds the target site, but does not cut either strand of DNA. Traditional Cas9 regulators In addition... then be fused to effector domains such as transcriptional activators or repressors. These fusion proteins...loci within a single cell. One might imagine transfecting cells with both the nuclease-positive and nuclease-null...Consequently, fewer genetic tools for targeted transcriptional and epigenetic regulation have been validated...inexpensively generate a potent Cas9-based transcriptional regulator without making any functional changes...

...summary per batch. Ranking guides for HDR in-depth In other tools, guides are ranked by specificity of ...genome by eye. We came up with a ranking formula that does just this: Rank = (compressed CFD score) * (gaussian...interactions (e.g., scores >80) are transformed to 100. This transformation (illustrated in the graph below...CFD scores (which range between 0 and 100) are then converted to a weight (which ranges between 0 and 1)...These values (ranging from -40 bp to 40 bp) are also converted to a weight (which ranges between 0 and...employs biological best practices. For example, it ranks guides by CFD score, and it mutates HDR donor templates...cut-to-insert distance) This gives each guide a score ranging between 0 and 1. The cutting frequency determination...

...contacts that bring apposing membranes together and can wrap the membranes into stacked structures including...dimerization.   1. Transmembrane fluorescent protein fusions Transmembrane proteins, i.e. receptors...receptors and transporters, are integrated into lipid bilayers. Fusion of an FP to a transmembrane protein significantly...Green Fluorescent Protein (GFP/EGFP) for a transcriptional reporter or as a general cytoplasmic label ...making FPs monomeric, yet many investigators remain ignorant of FP dimerization. Equally problematic, several...they cannot tumble in 720°, i.e. they are on a membrane or in an organelle, then their effective concentration...dimerizing fluorescent proteins is illustrated. 1) Membrane proteins fused to FPs that dimerize can form dynamic...

...activation of transcription in bacteria by co-opting σ54-based transcription. Transcription from σ54 promoters...enables researchers to maximize the chance of transducing a only a single gRNA per cell. The viral DNA ...genomic DNA. However, some cell types are poorly transduced by lentivirus, and thus screening with available...not possible. Primary murine T cells cannot be transduced by lentivirus and have been difficult to manipulate...Yusa Lab. This retroviral library was used to transduce T helper type 2 (Th2) cells from mice that constitutively...screening of cells that are refractory to lentiviral transduction, now enabling forward genetic screens in cells...they develop rapidly, they are small, and their transparency allows for whole organism imaging. They created...

...containing transcription factor binding sites can increase transgene expression and transduction efficiency...as the gene that they package. In a transfer plasmid, the transgene is flanked by two ITRs, and the ITRs...vectors, modifications can affect gene transfer efficiency and transgene expression (Zhou et al. 2017). For...these mutations can affect transduction efficiency and subsequent transgene expression. Replacing the ...nucleotide sequences that flank the ends of the single stranded DNA genome of AAV in the native virus. Since the... the feature its characteristic T-shape. The arrangement of the B-B’ and C-C’ palindromic sequences determines... reduce the packaging efficiency but increase transgene expression (Zhou et al., 2017). The most common...

...changing its double-strand cut to a single-stranded nick. In order to produce a double stranded break, two sgRNAs...place on the free strand of DNA and converts a C to U without producing a double strand break.   Prime... target DNA. The reverse transcriptase polymerizes DNA onto the target strand based on the sequence of...targeting dCas9 to transcriptional start sites was sufficient to repress transcription by blocking its initiation...approach is that two sgRNAs targeting opposite strands must be available in moderate proximity to each...desired edit adjoined to the sgRNA, a reverse transcriptase, and a nCas9. While this system has been shown...been exploited to localize factors such as transcriptional activators and repressors to target genes. ...

...RNA polymerase III transcript, the sgRNAs were encoded on the lentiviral minus strand in the 3’ LTR. These...HeLa cells transduced with SARS-CoV-2 nucleocapsid protein (Plasmid #141391-LV [+N]) or membrane protein ...high microglia transduction efficiency for the indicated ssAAV5s. Figure adapted from Serrano et al. 2023... with tRNAs, which are cleaved out of the RNA transcript by endogenous processing enzymes (Figure 1). ...including modifications to the Cas9 or reverse transcriptase enzymes, inhibition of host cell proteins, and...microglia are notoriously resistant to viral transduction. Through in vivo screening of human promoters...scAAV vectors have limited size compared to single-strand AAV (ssAAV). To maintain the packaging capacity...

...engineered combinations of transcriptional activators to induce gene transcription at genomic sites targeted...p65-HSF1 transcriptional activator to each genome-bound dCas9 (see Figure 1a for the transgenic construct...different untranslated or coding regions. Typically, each gene isoform has a distinct transcriptional start...investigation. Co-transduction of primary rat neurons with dCas9-VPR and sgRNAs targeting either transcript variant...activation (CRISPRa) tools for neuroscience, including transgenic mice, neuron-optimized viral vectors, and high-throughput...with the development of a Suntag-p65-HSF1 (SPH) transgenic mouse (Zhou et al., 2018). This mouse expresses...CRISPRa to selectively target these different transcriptional start sites in order to better understand isoform-specific...

...structures in RNA transcripts Effects on transcription (remember, it’s not all about translation - e.g. codon... you cannot choose among the possible codons at random and expect your sequence to express well in any...therefore may need fewer resources for the entire translation process. For example, during high growth rate... tRNAs. For example Tuller et al. found that translation efficiency is well correlated with codon bias...tRNAs, different regions of the protein may be translated at different rates. The ribosome will move quickly...the ribosome stalls, this may give the swiftly translated regions a chance to fold properly. For example...codon preferences online). Even if the gene is translated, it may not function properly. This is the result...

...small, double stranded RNA molecules that degrade target mRNA transcripts. When transiently introduced, ...fused to a transcriptional repressor to your gene of interest. It will inhibit transcription instead of...of introducing a DNA double strand break. Transient introduction of CRISPRi or inducible expression (more...LoxP vector.   Turn it off for a little while Transient suppression of expression is faster and easier...expression. Addgene has a dCas9 KRAB plasmid for transient knockouts!  Turn it on Turn it on forever Permanently...for integration at a specific genomic site, or randomly integrated (once or many times) into the genome...that, too! CRISPR activators (CRISPRa) are transcriptional activators fused to a dead Cas9. The system...

...and GFP reporters, as well as multiple transient and transgenic systems, Sahoo et al. showed that gene...commonly used 35S promoter. Transgenes are also efficiently inherited in both transgenic tobacco and Arabidopsis...tumor-inducing” or Ti plasmid, which allows it to transfer genetic material into the host plant genome. Scientists...Scientists have worked to optimize this system for gene transfer, studying the stability of modified Ti plasmids...a new and versatile binary Ti vector for both transient and stable gene expression applications in plants...borders specify the region of DNA that will be transferred to the plant, similar to viral vectors’ terminal...binary vectors are quite large (>10 kb.) Since transformation efficiency is inversely proportional to size...

...non-exclusive gene silencing pathways: transcriptional and post-transcriptional gene silencing (PTGS). In either...silence the target gene(s) either transcriptionally or post-transcriptionally. In plants, different dicer-like...where approximately 20-30 nucleotides of double-stranded RNA (dsRNA) results in downregulation, or “silencing...either pathway, short double-stranded RNA fragments (siRNAs) are produced from long dsRNA via an RNase III... through targeted mRNA degradation, blocking translation, or chromatin modifications of complementary ... Figure 1: Schematic of an RNAi-inducing transgene. Typically, the desired target of RNAi is expressed...alternative start codons, and/or the phenomenon of transcriptional adaptation (Smits et al., 2019; Sztal and Stainier...

...affiliated TniQ protein recruits transposition cofactors. The transposon proteins integrate a payload sequence... like we finish writing about one dual CRISPR-transposon system before another exciting new advance emerges...combined with the large sequence capacity of transposons open whole new worlds to explore. The newest ...called evoCAST — an evolved CRISPR-associated transposase (CAST). Back up a second. What are CASTs? CASTs...bacteria. In the course of evolution, Tn7-like transposons captured and repurposed nuclease-deficient CRISPR-Cas...Conveniently, the process doesn’t cause double-stranded breaks and can deliver large sequences and even...Cascade shown) combines with a donor template and transposon proteins (Tns) to integrate large sequences into...

...signal downstream of the transcription start site will terminate transcription of most of the ncRNA (Figure... coming, right? Non-coding RNAs (ncRNAs) are transcribed but do not encode proteins. They play important...Using a single guide RNA (sgRNA) to create a double-strand break works well because very small deletions and...sequence can completely disrupt expression of the transcribed ncRNA (Jiang et al. 2014).  Simple knock-out ...lncRNA regulatory elements that are crucial for transcription and downstream function. Rather than attempting...the lncRNA locus, scientists can mutate the transcription start site, splice sites, exons, and/or promoters...the removal of exons. Disrupting promoter or transcription start site function would also help to reveal...

...the zeptomolar range while One-step-SHERLOCK has a sensitivity in the femto-aptomolar range. For more details...are designed, each binding to one strand of the reverse-transcribed target sequence that are present close...reverse transcription. This is then amplified by PCR using Taq Polymerase and then transcribed to provide...LED and an orange filter to detect fluorescence upon cleavage. Highlights: Reverse-transcribed RNA samples... Suresh from Iowa State University. Since its appearance, SARS-CoV-2 has spread to almost every part of... milliliter can be detected – in the zeptomolar range Time: ~1 hour Equipment required: 37°C water bath...amplified. RPA reactions also contain reverse transcriptase to produce cDNA. Isothermal amplification is...

...proteins to process a double-stranded intermediate into the single-stranded genome that's then packaged...viral replication; and 3) the transfer plasmid which encodes a transgene of interest that’s packaged into...XmaI restriction sites). Another way is to transform AAV transfer plasmids into bacterial strains, like NEB...Overview: The first step in packaging AAV is transfecting HEK293 cells with AAV packaging plasmids. The... into the virus. Two to five days after transfection, the AAV-containing cells and media are harvested...more details. Pro tips I’m not going to lie, AAV transfer plasmids are kind of a pain to work with. These...structures that are prone to deletion from the transfer plasmid. The good news is there’s a few simple...

...Transcription factors (TFs) act as the gate keepers for gene expression, turning transcription on and... regulatory network. The transcription atlas and library The transcription factor library The human...lentiviral particles and used to transduce human embryonic stem cells. Transduced stem cells were differentiated...approximately 1,800 TFs, with >3,500 unique isoforms transcribed from these sites. The Zhang lab generated a comprehensive... contribute to differentiation, the Zhang lab transduced human embryonic stem cells with the viral TF ...differentiation outcomes as well as global transcriptional changes. Using this road map, and what was ... uncharacterized TFs is a useful finding. Transcriptional program changes from this screen successfully...

...the transient cotransfection of the mARG plamsids into HEK293T cells using a chemical transfection reagent... These plasmids can be used directly in transient transfection in rtTA+ cell lines (such as HEK293-TETOn...into the genome by cotransfecting them with a plasmid expressing PiggyBac transposase. In rtTA-cell lines...engineering a cell line, cotransfect the mARG cassettes 2:1 (A:N-V) with PiggyBac transposase and outgrow the ... of replication (p15A) worked best under a wide range of inducer concentrations (0.1-1% L-arabinose) without...proteins. Excess gvpA plasmid added into the transfection mixture in HEK293T cells gave drastic improvement...improvement in expression. Stabilizing the gvpA transcript with WPRE reduced the number of copies of gvpA ...

...is codon usage important? Translation and protein synthesis relies on transfer RNAs (tRNAs). tRNAs bind...rate of translation and thus protein expression. In fact studies have shown that translational efficiency...on an organisms codon usage bias to increase translational efficiency and thus protein expression without...mammalian cells. This was the first codon optimized membrane protein expressed in mammalian cells. In this ...conformation, folding and stability, change post-translational modification sites and even affect protein function...Mauro and Chappell, 2014). Different rates of translation by different tRNAs, including those that exhibit...actually be critical for determining the rate of translation. The ribosome may slow and pause during elongation...

...concentration of FLEx vectors guarantees that most cells are transfected with all labeling plasmids and...PMCID: PMC2593125. 2. Buchholz, F, P O Angrand, and A. Francis Stewart. 1998. “Improved Properties of ... sites can lead to excision, inversion, and translocation of DNA in a similar fashion as between loxP ... has a temperature optimum of 37 °C (Buchholz, Angrand, and Stewart 1998). FLPe’s performance was further...in FLPO, which shows similar efficiency to Cre (Kranz et al. 2010). These differences in recombinase efficiency...-LDB domains of the fusions allowing Cre/FLP translocate to the nucleus where they exert their recombinase...brain? Cells of a specific brain area can be transfected by in utero electroporation. To achieve sparse...

...Importantly, they allow transient delivery, enabling transgene-free genome editing (no transgene integration), ..." (one for each strand), while TnpB, like Cas12a, uses just one to cleave both strands. Similar to how...lab recently harnessed TnpB from Deinococcus radiodurans (Dra2TnpB) to create a genome editing system ...also need a short sequence motif called a TAM (transposon-associated motif), which is positioned like Cas12a...species. For instance, the TnpB from Deinococcus radiodurans (Dra2TnpB) recognizes the TAM sequence 5′-TTGAT...parts: a structural scaffold derived from the transposon's right-end element, and a 20-nucleotide (nt) ...downstream of the TAM, creating staggered double-strand breaks like those created by Cas12a. Applications...

...neurons! Neurons are really cool because they can transmit information, for example, from one of the 180 ...associated virus (AAV) shows relatively low retrograde transport efficiency in most serotypes. Some viruses (e....e.g., rabies) naturally exhibit retrograde transport and can therefore perform retrograde tracing, but...efficiently infects neurons, directs high-level transgene expression, and has retrograde functionality to...genetically controlled function. By using AAVrg in Cre transgenic mouse lines, you can access and manipulate an...from the hippocampus to Broca’s area would be transduced (Fig 3B). These specific neurons could then be...requires high titers for efficient retrograde transport. For recombinase-dependent vectors, this can cause...

...through transformation, so the cloned vector must be ligated. But in LIC, longer single-stranded overhangs...T4 polymerase (left branch); by annealing two separate PCR products (middle branch); or by incomplete ...step to retain products with single-stranded overhangs (right branch). The mixed PCR and incomplete PCR...overlapping regions of single-stranded DNA. But as long as the double-stranded region is long enough to hold...nicks easily repairable by the bacteria after transformation. LIC is a reliable cloning method, but it is... sequences (20–60 bp) on either end and single-stranded 5′ overhangs… but it doesn’t really matter if ...recombination intermediate to bacteria, so it can be transformed as-is into cells. The cells then get to work ...

...the opposite strand). ABEs make an A to G change (or a T to C change on the opposite strand). Both are ...place on the free strand of DNA and converts a C to U without producing a double strand break. UGI = uracil...Repair CRISPR 101: Cytosine Transversion Editors CRISPR 101: Any Base Transversion Editors Learn about base...repair (HDR), the pathway for resolving DNA double-strand breaks (DSBs) preferred by researchers. Compared...are base transition editors: converting one purine base to the other or one pyrimidine base to the other...other. Figure 1: Base transition editors catalyze purine-to-purine or pyrimidine-to-pyrimidine base... indicate the change introduced to the opposite strand following base editing and DNA repair. Black arrows...

...therapies and in vivo laboratory experiments ranging from transient gene expression in a particular cell type... Figure 2: DNA rescue and transduction of a conventional single-stranded AAV (ssAAV) and a self-complementary...genome conversion in the transduced cell nucleus, the single-to-double stranded conversion of the DNA goes...structure can readily fold into transcriptionally active double-stranded form through intra-molecular annealing...Adeno-associated virus (AAV) is a single stranded, Parvoviridae DNA virus, packaged in a non-enveloped...hijack host cell machinery, and synthesize a second strand of DNA before expression of their cargo can begin...integrate into the host genome and therefore the transduced gene is episomal and does not divide and replicate...

...for broad or narrow range of microbial species. These also contain the RP4 transfer origin as well as a..., the ability to detect mRNA transcripts of toxins and other transcripts of interest is a benefit of this...the organisms present while sequencing of all transcripts gives clues into what the microbes are doing....followed by a reporter gene. The hairpin blocks translation by sequestering the RBS, but binding of the “...microbial community) reveals the RBS and allows translation of the reporter gene resulting in GFP signal....lacks genetic tools is the bee microbiome. Nancy Moran’s and Jeffrey Barrick’s labs at The University of... RSF1010 origin, an IncQ origin with broad host-range (Meyer, 2009). The vector retains the oriT for delivery...

...inducible gene expression. Transgenic and knockin mice As the name suggests, transgenic and knockin mice contain...their genome. While transgenic mice have the new gene construct incorporated randomly into the genome, knockin...Conversely, outbred strains were originally derived from random crosses between mice of uncharacterized backgrounds...several types of genetically engineered mice: transgenic mice, knockout mice, and mice with conditional...construct inserted at a specific site. To generate a transgenic mouse, microinjection of DNA or infection with...Non-homologous recombination incorporates this DNA randomly into the mouse genome. In contrast, knockin strains...animal for gestation. Nucleases that induce double-stranded DNA breaks, such as ZFN and TALEN, can used to...

...generally are not transmitted to adjacent neurons via synapses (i.e., transmitted transsynaptically). This is ...retrograde transmission (Etessami et al., 2000, Wickersham et al., 2007). Thus, RABV transsynaptic spread ...replicate. Modified Rabies Virus: Converting a transsynaptic neuronal tracer into a monosynaptic (first order...subsequent cycles of viral replication and retrograde transmission occur, spreading the virus through higher and...higher order neural connections. While this transsynaptic spread is useful for neuronal tracing, it has...-dependent TVA to Cre-expressing neurons in a transgenic mouse line. By doing this, RABV expressing the...necessarily express EnvA unless it is supplied in trans. Monosynaptic RABV spread can be achieved by modifying...

.... They can be targeted to the plasma membrane to measure membrane potential and can also be directed to... the membrane), the voltage sensing component (i.e., rhodopsin naturally resides in the membrane), or ... of voltage gated ion channels all depend on transmitting and sensing electrical potential. Thankfully... want to monitor the voltage change across the membrane of cardiac cells. Similarly, neuronal signaling...propagated through multiple cells to do things like translate perceived signals to physical actions. Understanding...rhodopsin proteins were initially evolved for ion transport, not fluorescence, they are not nearly as bright...sensors contained a FP that localized to the plasma membrane due to the addition of several targeting domains...

...discovered in positive single-stranded RNA viruses to allow for the efficient translation of multiple genes from...Elements Translation in eukaryotes usually begins at the 5’ cap so that only a single translation event ...scientists use a multitude of techniques including co-transfection of two or more plasmids, the use of multiple...Unlike promoters which will create unique mRNA transcripts for each gene that is expressed, multicistronic...your gene as they are both derived from the same transcript. Of course multicistronic vectors do not have...from a single transcript. The two strategies most widely incorporated into plasmids for research purposes...Entry Site (IRES) to allow for initiation of translation from an internal region of the mRNA. In the ...

...recent paper, "Engineering Complex Synthetic Transcriptional Programs with CRISPR RNA Scaffolds." Adam has...visualize the concepts discussed in the paper. The transcriptional control of multiple loci is deftly coordinated...cerevisiae with few techniques that are easily transferrable across host species or metabolic pathways. A...CRISPR-based RNA scaffolds to mimic natural transcriptional programs on multiple genes. Modifying the ...synthetic RNA scaffolds that can recruit various transcriptional regulators (Zalatan et al. 2015).  These scaffolds...nucleotide sequence then subsequently recruit a transcriptional regulator to modulate expression at that locus...expression levels dependent upon the type of transcriptional regulator or the number of designed scaffold...

...:     Cells must be amenable to transfection or viral transduction     Appropriate promoters must be...expression     The requirement for Cas9 transcription and translation delays editing What are Cas9-gRNA ... studies in cells that are difficult to transfect or transduce. While using Cas9-gRNA RNPs may require...site of interest. This is commonly performed by transfecting cells with a plasmid, such as PX459, which encodes... often be used in cells that are difficult to transfect, such as primary cells. Using RNPs can also alleviate... are detectable at high levels shortly after transfection, and are quickly cleared from the cell via protein...from Alex Schier’s Lab. gRNAs can be in vitro transcribed from ssDNA, which can be generated by commercial...

...contains 11 β-strands and one central ɑ-helix that can be split into 3 parts: (1) β-strands 1-9 (β1-9) and... and the one ɑ-helix, (2) β-strand 10 (β10), and (3) β-strand 11 (β11). Each part contains components ...developed, such as the Forster resonance energy transfer (FRET)-based executioner caspase reporters. In...need a protease reporter that has a large dynamic range and high brightness. The Shu Lab from UCSF designed...However, the lab wanted to increase the fluorescence range to better study the spatio-temporal dynamics of ...GFP (GFP1-9 and GFP10-11) by flipping a single β-strand of GFP, hence the name FlipGFP. FlipGFP increases...from β1-9 they will rapidly recombine with β1-9 strands nearby resulting in fluorescence within tens of...

...humanized wt Cas9, dCas9 (transcriptional repressor), and dCas9-VP64 (transcriptional activator)-containing...Multiplexing from a single transcript You can also multiplex gRNAs via a polycistronic transcript. Rather than being...being transcribed from different promoters, the gRNAs are transcribed together and are flanked by specific...simultaneously. Vectors for both transient expression and Agrobacterium-mediated transformation are available. For ...system for processing a continuous multi-gRNA transcript, each gRNA must be expressed from its own promoter.... But that doesn’t mean you have to clone and transfect multiple promoter-gRNA constructs in order to ... Cas9, D10A nickase mutant (Cas9n), dCas9 (transcriptional repression), and Fok1-dCas9 (dimeric nuclease...

...but also a biological transposon system. Sleeping Beauty is a synthetic transposon system that can be used...reconstructed a transposase system based on the naturally mutated and non-functional transposase gene found...resurrecting this transposon system? Scientists can use Sleeping Beauty transposons as a non-viral system...Addgene recently moved to our brand new location, with more space to grow and help scientists share resources...you can use to study these genes! Groucho, a transcriptional co-repressor The first conference room, that...Addgenies, was named after the protein Groucho, a transcriptional co-repressor important in development. Groucho.... It binds to a specific set of DNA-binding transcription factors to repress the expression of genes primarily...

...required for hok translation. Here, Mok and Hok translation is coupled. Translation from the Shine-Dalgarno...more stable than the methyltransferase. When a plasmid is lost, the methyltransferase is degraded and/or...systems have been used as biological tools for a wide range of applications in molecular biology or in vivo ...ccdB gene die off and a higher percentage of transformants contain the ccdB-less construct. Other toxin-antitoxin...composed of restriction endonucleases and methyltransferases. Restriction endonucleases cleave DNA at ...specific nucleotide sequences. To counter this, methyltransferases add methyl groups to bases at specific nucleotide...or diluted as the cell grows resulting in the appearance of unmethylated sites in the chromosome. The ...

...opportunity to discover and evolve your transferable skills. Transferable skills are the non-technical skills...employee who's coming directly from grad school. The transition has been a bit rough for her (3 weeks in and ...there are any resources that can help with the transition.” The lab-to-desk-job culture shock is a real...scientist colleagues have as well. Making the transition from the bench can be jarring for the unprepared...resources out there for scientists making this transition. In this installment of the Addgene career guide...brief period of time, the only, person to know a brand new fact is often enough to keep a scientist going...formulate hypotheses and conduct experiments. You transfer liquids between tubes, look through microscopes...

...different ligands, including hormones, neurotransmitters, photons or odorants, to elicit downstream signaling...seven-(pass)-transmembrane domain receptors, due to the seven alpha helices that transverse the cellular...cell membrane. Heterotrimeric G proteins are important molecular switches in signal transduction pathways...fusion to a receptor-transcription factor fusion. TEV protease cleave the transcription factor from the receptor...initiating signaling pathways in inflammation, and neurotransmission. These receptors can be activated by an array...cellular membrane. The receptor recognizes the appropriate ligand to activate the G proteins. It is estimated...parallel receptorome expression and screening via transcriptional output - TANGO) (Kroeze et al., 2015). PRESTO-TANGO...

...increases when target DNA strand separation is stabilized (2,3). Stable strand separation is maintained...and therefore destabilize basal strand separation. This decreased strand separation combined with the weak... Figure 1: Strand separation and target DNA binding to the Cas9 nuclease. Stable strand separation is...the non-target strand and the HNH/RuvC groove, and interactions between the target strand and the gRNA....interactions : 1. Interactions between the non-target strand and a positively charged groove formed by the Cas9...nuclease domains 2. Interactions between the target strand and the gRNA (see figure below) Because off-target...by a technique for detecting genome-wide double strand breaks (BLESS), revealed that these mutants do ...

...new class of sensors for neuropeptides and neurotransmitters. Motivated to develop a biosensor for oxytocin...devised a “Modular fluorescence unit fused with TRansmembrane region-to-IntrAcellular loop linkers” (MTRIA...sensors based on GPCRs recognizing various neurotransmitters, neuropeptides and other ligands - almost ...kinetics and localization for imaging synaptic transmission. bioRxiv 2022.02.13.480251. doi: 10.1101/2022.02.13.480251...2022.02.13.480251 JEDI-2P voltage sensor The François St-Pierre lab and colleagues have developed a new...they observe fluorescence localizes to the plasma membrane only when the RTK is activated and the ITAM phosphorylated...relies on a change in localization to the cell membrane and not a direct change in the fluorescent properties...