...fluorescent proteins (FP) require external light activation, you can’t use fluorescence to monitor processes...light used for excitation can cause unintended activation of the optogenetic system. Biosensors based on...external light source. Therefore, especially thick samples may not be appropriate for use with Nano-lantern...Resources on the Addgene Blog Learn about Photoactivatable Fluorescent Proteins Check Out Fluorescent...

...structure can readily fold into transcriptionally active double-stranded form through intra-molecular annealing...expression patterns of AAV8 vectors with broadly active promoters from subretinal injections of neonatal...episomal and does not divide and replicate in the same way the host DNA does. Because of this, fast dividing...McCarty, D. M., Paul E. Monahan, and Richard J. Samulski. "Self-complementary recombinant adeno-associated...

...recombinant adeno-associated viruses for Cre activated and inactivated transgene expression in neurons.  PubMed...ability to conditionally turn off one gene and activate another. In the field of mouse genetics, FLEx ...between the sites. If a DNA sequence is floxed in the same orientation, the SSR will excise the DNA fragment...reporter with one loxP site and one lox511 site in the same orientation, and placing the additional loxP and...

...scientists and non-scientists alike, need to stay active and involved if we wish to make positive change...chair of the postdoctoral engagement committee and active member of the university's Postdoctoral Association...this year I had never taken part in any public activism. One of my favorite signs from the march read ...Deanna McLafferty is an editorial assistant and activist. Washington, DC, USA 38.9072°N 77.0369°W – Mark...campus speakers thoughtfully reminded us to use the same critical lens on our scientific community as we ...well-publicized, we might encounter hostile anti-science activists. However, I must say that I was supremely proud... nor would you mention my small error and at the same time pass over in silence his very large one” (http...

...subsequently destroyed by Argonaute-catalyzed nuclease activity or their translation disrupted through physical...the appropriate plasmid and dsRNA expression is activated through the  induction of T7 polymerase encoded...Students can produce DNA templates via PCR off of the same plasmid and use these to create riboprobes for in...WISH protocol involves preparation of the tissue sample, several washes, and temperature changes to visualize...

...could in theory promote off-target binding. The activity of SpRY was validated in double stranded cleavage...assessment of the PAM compatibility and editing activities of Cas9 variants. Nucleic Acids Res. 2021 Sep...& Walton et al). Additionally, SpG Cas9 with the same PAM preference of NG was isolated by the Kleinstiver...several bps away from a desired site does not have the same devastating impact on efficiency as 10-15 bps away...

... derailing the show. The moderator must be very active at all times keep things moving and to prevent ...interesting to the audience. It is a bit of an art, but active moderation gets easier with practice. Up the audience...answers uninteresting, the panel members all say the same thing and one panel member dominates the discussion... for an intimate event, put the panelists on the same level as the audience. In my experience the best...

... the known pathway that were essential for the activity of different ligands. Such knowledge could be ... to other nucleases as well as inhibitory and activating approaches.  Many thanks to our guest blogger...G., et al. "Optimized sgRNA design to maximize activity and minimize off-target effects of CRISPR-Cas9...validation screen using selected gRNAs extracted from the same or another library. These different approaches could...Listed you can also easily extract gRNAs for the same gene from several libraries, and identify those ...

...enough regulatory elements, CRISPRs, repressors, activators. We’ve been putting together very large toolboxes...the amount of transcriptional and translational activity a cell has and control how much it taxes the resources...problems. You couldn’t do it without repeating the same guide RNA twice. The second problem was the shape...

...interest wanes off such that attendees are not actively participating, it may be time to consider ending...clubs where participants are passive rather than active and the meeting is often driven by one or two participants...itself A journal club should aim to have many of the same characteristics as a successful meeting: a clear...through the paper figure by figure. Some follow the same figure by figure organization with a different person...stay abreast of current trends in the literature, sampling a breadth of work from a field (perhaps even two...outside of one's own field in a structured and interactive way. Like any meeting, to be successful in serving...

...site of incorporation (Figure 1d). Fluorogen activating proteins (FAPs) – These modified antibody fragments...William Perkin. His discovery set off a flurry of activity and the majority of the classic small molecule...removing excess dye from the sample, which is particularly important for samples where washing is difficult...small-molecules: red-shifted spectra, ion sensitivity, photoactivation, etc. These impressive advances lead to an ...reason is something called “the photon budget”: each sample has a limited number of fluorophores and each fluorophore...of information one can extract from a biological sample is wholly dependent on the photon budget and pushing... desperate to extract a few more photons from a sample. Chemical fluorophores can be substantially brighter...

...directly, albeit weakly, to K-Ras and inhibit its activation. The next step may be to follow up on these promising...molecules that bind to K‐Ras and inhibit Sos‐mediated activation." Angewandte Chemie 124.25 (2012): 6244-6247....brake pedal that does not work. Both cases yield the same result – uncontrolled movement of the vehicle or...transcript most commonly found across all TCGA tumor samples is represented, making the results of any such ...screenshot below). Currently, the page includes interactive maps for the Ras, mTor, and p53 pathways. The...

...only be aware of the potential for bias but to actively look for and consider bias as a confounding factor...s imperative for scientists to be aware of and actively use tools designed to help reduce or eliminate...sequences of guides across 8 CRISPR libraries. Samples are divided into those of African ancestry(orange... on these and any other CRISPR screens using the same reference genomes and libraries. Further complicating...

...you up to speed easier and faster when you’re an active participant in the training. In this blog we will...assistance at some point during the procedure. Be an active partner Everyone’s learning style is different,... a saying that if the sample prep is poor, then the data will be poor. The same is true for training in...you remember where the equipment, reagents, and samples are. Do: Ask your bench mentor if they have time...

...around 1,000 active makerspaces around the world. Hackerspaces.org maintains a list of active spaces near...data is freely available, and the collection of samples and sequencing is being organized and funded by...

...PE1 variants with RT mutations known to increase activity, enhance binding between the template and primer...prime editor sequence to improve expression and activity In addition to the numerous innovations that streamline...mutations in Cas9 previously shown to improve nuclease activity (Chen et al., 2021). In a system as complex as...R221K N394K Cas9 mutations Improve Cas9 nuclease activity ~ ~ ~ ~ ✓ ✓ ✓ Evolved reverse...likely because both innovations aim to solve the same problem. Both approaches are effective, but users...

...nucleases (such as Cas3) or have its own nuclease activity. Instead, it forms a novel co-complex with the... (Vo et al., 2020). VchINT was also shown to be active in various gram-negative bacterial hosts of interest...seven required genes and the CRISPR RNA under the same promoter, and a donor plasmid (pDonor) encodes the...immunity, whereby multiple proximal insertions with the same transposon exhibit reduced efficiencies (Vo et al...

...Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening - An excellent protocol paper by the...Genome-scale CRISPR-Cas9 knockout and transcriptional activation screening. Nat Protoc. 2017 Apr;12(4):828-863...plasmids, all of which need to be represented in the same proportion after amplification as they were prior...visualization) that confirm diversity of the library sample, and any other information or scripts provided ...

...modification with Homology Directed Repair (HDR), CRISPR activation or repression, transgenic mouse line creation...deliver Cas9 variants, such as a transcriptional activator dCas9-VPR, if these variants are fused to the ...detected in cells electroporated with RNPs using the same guide. Nanoblades also had lower off-target effects...

...Zhang, Y., & Ke, A. (2024). Exploiting activation and inactivation mechanisms in type I-C CRISPR-Cas3 for...called CasPEDIA, with information about enzyme activity, experimental conditions, and more. Type II Type...enzyme, with adaptations for increased fidelity, activation/repression, high precision edits, and more. ...data. In some cases, CRISPR systems that are in the same class and type don’t share sequence similarity but...between two types, the Cas effector may have the same name but be completely different! They are merely...but it’s suspected that they don’t carry out the same types of adaptive immune functions that other CRISPR... 2000–2012. https://doi.org/10.1093/nar/gkz1197 Samai, P., Pyenson, N., Jiang, W., Goldberg, G. W., Hatoum-Aslan...