... CRISPR-Cas technology is constantly evolving. Variants of Cas proteins can be used for genome editing...In this study, the labs found that anti-CRISPR prevents Cas9 activity in HEK293 by reducing editing from... AcrIIC2 interactions with Cas9 interferes and prevents the correct assembly of the crRNA-Cas complex ...binds to the HNH endonuclease domain of Cas9 and prevents target DNA cleavage (Harrington et al., 2017)....   Five ways to anti-CRISPR proteins in your experiments 1. Reduce off-target effects Prolonged Cas activity...shut off Cas activity? Using inhibitor timing experiments, the Doudna lab found that at least 50% of on-target...vectors. Having a way to shut off CRISPR-Cas systems prevents the off-target effects and cytotoxicity that result...

...T-cells were transferred back into the infants. Almost all patients have achieved persistent immunological...To date, gene therapy has been used to treat 10 infants with this disease. To do so, their T-cells were...administration of rAAV2 carrying the human gene RPE65, patients with clinical diagnosis of LCA and mutations in... in RPE65 (n=3), showed significant improvements in vision over a period of 1 to 3 years – the treatment...Anti-AAV antibodies developed by the immune system – patients’ own immune systems can develop antibodies that...delivery tools that are under consideration, recent advancements in various AAV technologies have vastly improved...

...the following four components into 293T cells. Figure 1: The key components of Nanoblades. Image... deliver Cas9 variants, such as a transcriptional activator dCas9-VPR, if these variants are fused to ...CRISPR applications. Let’s take a look at the key components of Nanoblades, how their editing efficiency and...for HDR editing and mouse-model generation. Key components of Nanoblades Nanoblades are a Cas9 RNP delivery...transgenic mice by microinjecting zygotes with CRISPR components. However, this sometimes requires intracellular...

...mortality determined by nutrition during parents' and grandparents' slow growth period." European Journal... histones, the more compacted it becomes. This prevents the cellular machinery from physically interacting...were experienced in a period of life when the grandparents’ gametes were developing, an epigenetic change...intellectual and developmental disability, jerky movements, and spontaneous laughter or smiling, whereas ...enabling faster diagnosis and treatment and improving patients lives. References 1. Ong-Abdullah, Meilina, et...Biosensors Read Our Tips for Using FRET in Your Experiments Post Read Other Blog Posts about Fluorescent ...

...transposase (CAST) that can insert large fragments of DNA. The V-F variants, which are among the smallest Cas...on main function and composition; subtypes and variants are typically based on species of origin. There...currently two classes, six types, 33 subtypes, and 17 variants. Figure 1: Overview of the CRISPR classification...mechanisms, with the biggest differences being the components present in the Cascade complex. Figure... Cas2, which are critical for inserting short fragments of foreign DNA into the CRISPR array. Lastly, ...CRISPR–Cas systems: a burst of class 2 and derived variants. Nature Reviews Microbiology, 18(2), 67–83. https...Insights from CRISPR Loci in Integrative Conjugative Elements of Acidithiobacillia. The CRISPR Journal, 4(5)...

... to teach students about basic plasmid biology Use or CRISPR 101 eBook to introduce students to this amazing...an open-access journal that enables high school students to publish peer-reviewed scientific research. ... and hear from a fantastic team of high school students who recently submitted to the journal with funding...Investigators. Chapter 2 - Getting high school students and teachers interested in JEI (the importance...audience and their current readership. Encouraging students to do science anywhere.  Chapter 4 - JEI's mission...

...are used to repeating experiments to confirm their findings. But no scientist wants to repeat an experiment...errors increases. In addition to human error, fragments of DNA from the helper plasmids or the cell genome...serotype of the packaged DNA and look for potential contaminants. The analysis is a 2-step process using Geneious...software looks for and quantifies the presence of recombinants in recombinase-dependent sequences. Recombinase-dependent...powerful tool that can be used to identify DNA contaminants in AAV preparations and provide information ... AAV vectors for your research. Please use the comments section below to let us know if you have any specific...

...technology that allows precise modifications of DNA (replacements, small insertions, and deletions) without introducing...based on patterns observed from high-throughput experiments, where we can evaluate tens of thousands of designs...Figure 1). Through this method, we found key determinants of prime editing efficiency such as the length...   How to apply PRIDICT in prime editing experiments Prediction models only become useful once they...PRIDICT2.0) can greatly facilitate prime editing experiments, thus saving you time, sweat, and money on extensive...especially shines once large-scale prime editing experiments come into play. Nicolas Mathis received his ...efficiencies using sequence features and DNA repair determinants. Nature Biotechnology 2023, 1–11. https://doi.org...

...Thankfully, there is a tool to measure these electrical events! Voltage indicators are voltmeters for cells! In...chemigenetic sensors. These tools have two main elements: a voltage-sensing component and a fluorescent...voltage sensor with a genetic component. The VSD components have historically come from a variety of sources...squirts. This design structurally separates the two components of the indicator, which can make the kinetics...types of sensors can have a variety of different components, but all contain a FRET pair, a genetic component...Viral vectors are mechanisms by which genetic components can be delivered to cells both transiently and...covered. Happy voltage sensing!   Figure 3: Components of the types of voltage indicators reviewed. ...

...vectors in the brain and peripheral nervous system presents a special challenge to scientists. What works ...neurons (Figure 1). However, this one set of experiments didn’t provide any information on other important... of the virus to adjacent areas impacted the experiments we wanted to perform. It required a great deal...community concerns AAV vectors are complex biological reagents; many factors, like storage buffers and conditions...conducted to improve our Addgene Data Hub, 76% of respondents indicated they wanted to see more negative data...data they cannot explain, perhaps out of fear it points to flaws or gaps in their technical ability. But...and use this data to guide them in their own experiments. By providing DOIs, this information can be cited...

...lentiviral vectors! Lentiviral vector history and components In the early 1990s, researchers developed viral...and Pol, increasing the number of recombination events necessary to create a replication-competent virus...trial with a lentiviral vector. CD4+ T-cells from patients with HIV-1 were transduced with a lentiviral vector...sequence against the HIV-1 envelope. As of 2013, 65 patients had been treated with this therapy, which lowered...beta-thalassemia. Lentiviral vectors are promising agents for cancer immunotherapy, but this research is ... vectors contain a deletion in the 3’ LTR that prevents aberrant activation of nearby genes. These safer...experiment. Let us know how you use lentivirus in the comments section below! References 1. In vivo gene delivery...

... coli strains producing different pigments, and tasked the students to separate out colonies from each...second quadrant. The process continues until all quadrants are used, hopefully revealing single, isolated...incubation. The challenge is usually successful for the students. “They always get a few isolated colonies. I should...chromoprotein-expressing strains are easily amenable to other experiments to teach other aspects of molecular biology. ... do with the plasmids from Addgene is have the students transform them into different strains of E. coli...

...are going to be there – undergraduate students, rotation students, joining a new lab at any career stage... help clean up and/or make necessary reagents for the experiments. Unlike in lab courses, where everything...double-check you remember where the equipment, reagents, and samples are. Do: Ask your bench mentor if...everything is magically made up by helpful TAs or assistants, and sitting out on the bench waiting for you...with…now”. Ask for feedback so you can make adjustments. This could look like “What are some things I...

...minutes. While users can’t tangibly see these improvements, you are benefited with reassurance your plasmids...on stories during short periods of time called sprints. The team is mindful of the number of stories we...without impacting the overall work. Each of our sprints is themed and the stories in that sprint relate.... Third party libraries are reusable software components that can be incorporated into software. You can... can think of these libraries as ingredients to a recipe. It’s unrealistic to plant, grow, and harvest...cake depends on how you combine it with other ingredients.  For example, we use a third party library to...over again for every single software today that prints labels. These libraries in turn need to stay up...

...option, try the Biospace career events page. Biospace holds career events all over the country; sometimes... thereof) One of the biggest challenges facing students today is that many graduate and post-doctoral ... development training. Further, even for those students and researchers who do take the initiative to ...obviously important experience, but far too many students and young professionals will never attend a job...which are larger meetings offering resources for students seeking employment within industry or academia...using ResearchGate to see where other graduate students and post-docs from your institution have gone,...

...feeling out of their lab, but for many graduate students and postdocs, the people you see in your bay for... a lab. You formulate hypotheses and conduct experiments. You transfer liquids between tubes, look through...office transition? I’ve already provided a few counterpoints to some of the causes of culture shock above...it’s like to be properly onboarded. Most work environments outside of academia provide a formal training...probably because you don't have to worry about experiments taking longer than expected or dealing with unexpected... bring real scientists into schools to provide students with hands-on lab opportunities. You can get your...are there once you start looking for them. Acknowledgements Big thank you to Jessica Welch, Mike Lemieux...

.... This enhanced movement is thought to reflect elements of drug addiction and craving (‘seeking drugs’...approach against drug addiction. Anxiety tests Rodents have a natural preference to stay in dark areas...ones (risky). They also prefer to explore novel environments by initially moving along a wall (protected)...Taking advantage of these characteristic traits, rodents can be used to study anxiety levels in general,... driven by their motivation to explore novel environments, rather than being inhibited by fear/anxiety...test the efficacy of antidepressant treatment in rodents. In this test, an animal is placed into a tank ... of microRNA124a in the hippocampus evokes antidepressants-like effects in rats." Psychoneuroendocrinology...

...it's time to take a look at some of its parts. Elements such as the antibiotic resistance gene and the...polymerase binding site) and operators (response elements). Promoters are about 100 to 1000 base pairs long...RNAP can recognize and bind to specific promoter elements. This means that the promoter present in your ...various eukaryotic cell types (mammalian, yeast, plants, etc) require unique promoters and there is very... other regulatory factors: enhancers, boundary elements, insulators, and silencers; however, some "leakiness...typically include some combination of other promoter elements, and tend to be more tightly regulated. Common...ubiquitin gene Constitutive  Gives high expression in plants. H1 small RNA expression shRNA From the human...

...genome editing in plants While there have been many Cas nucleases developed for plants, it has been difficult...Encyclopedia of DNA Elements (ENCODE), a consortium aimed to identify functional elements in the human genome...specificity of multiple wild-type and engineered variants of Cas nucleases. In doing so, they also created... toolkit for Cas-mediated genome engineering in plants. Read the paper in bioRxiv Find the plant genome...

...and Harvard. Students also partner with independent career organizations to get the grants and programs...recently appeared in eLife: "A Fair Deal for PhD Students and Postdocs" by Dr. Henry Bourne, Professor Emeritus...proactive role in their non-bench training. Grad students at Washington University founded a successful ... enough. There are still university research departments that have no formal career resources for science...science trainees at all. For grad students and postdocs interested in taking matters into their own hands...