...how to prioritize but I  just added the “most important” tasks my schedule. This was bad for 2 reasons...expectations, answer any questions, and (most importantly) decide on a timeline. My experience is that ...your team’s success depend on it.  Equally as important is to be able to actively listen - this can be...for the best. Positive feedback is equally as important so don’t forget to recognize someone’s hard work...managers and I've learned from both. The former will empower you, keep you motivated and help you grow in your...

...generations. Figure 1: Mobile-CRISPRi's modular components. Image from Peters et al., 2019 with permission... Top: A Tn7 transposon containing the CRISPRi components and a plasmid with the Tn7 transposition genes...ability to synthesize diaminopimelic acid (DAP), a component of the cell wall. Once in the recipient cell, ...Bottom: An ICE element containing the CRISPRi components is transferred to recipient bacteria by bi-parental...experiments where maintaining library diversity is important, while lower transfer rates are only suited for...

...post was updated on Aug 20, 2020. What genes are important in your phenotype of interest? Many scientists...not entirely known. Identifying which genes are important for a phenotype can lead to a wealth of additional... and versatility. CRISPR/Cas9 consists of two components: the non-specific endonuclease (Cas9) and a single...library is a complex mixture. Pooled libraries are composed of many different gRNA-containing plasmids, each...strong evidence that a particular gene is actually important for a given phenotype. Each hit from the screen...

...experiment requires delivery of, at minimum, two components: the Cas9 protein and a guide RNA (gRNA) targeting... by commercial vendors such as IDT. These two components are then incubated together to form the RNP. ... account with these types of experiments. One important consideration is that the Cas9-gRNA RNP will be...scientists with an additional means to deliver CRISPR components to their experimental systems of interest. Using...it is likely that the use of RNPs will play an important role in its continued advancement. Andrew Hempstead...

... a Type I-F CRISPR-Cas system. The four major components required for RNA-guided DNA integration are (...sequences (R and L ends).  Figure 1: (A) Components involved in each type of CRISPR-Transposon system... proteins (TnsB and C). While requiring fewer components for RNA-guided DNA integration, Type V-K systems... promoters were used to express the necessary components in E. coli (Klompe et al., 2019). Recently, the...simple to use, users should keep aware of a few important details. For transposons to be specifically recognized...

...of the vector components are HIV-based? A7: No single plasmid contains all the components necessary to ...The components are divided as follows: Transfer vectors contain minimal cis-acting HIV components: LTRs...LTRs, PPT, RRE, and psi packaging signal. Viral components typically total <1.5kb, which is rarely more ...

...of production have also made aptamers an important component of high throughput assays with microarrays...diagnosis. As biosensors, aptamers hold a few important advantages over their amino acid-based counterparts...laser cleavage and an RNA aptamer that bound the compound malachite green (MG) to create a non-protein based... 4-hydroxybenzylidene imidazolinone (HBI)-like compound allowing it to fluoresce, the Jaffrey lab asked...

...published data. The importance of data sharing The reason this topic is important to my lab is three-fold...their own data? A distinct (perhaps similarly impoverished lab) might apply different or novel tools and...work this may entail.  So if access to data is important, what data should be included? We should recognize...deliver on well-meaning but impractical requirements imposed by publishers and funders. Clearly, it is in our...

...likely not a coincidence that the AAV genome is composed of single stranded DNA, which is particularly ...with G418-selected clones. However, there are important disadvantages to the AAV-based approach that must...systems. Ohmori et al 2017 use a dual vector system composed of SaCas9 and a gRNA in one AAV and a repair template...researchers have devised methods to deliver CRISPR components as ribonucleoproteins (RNPs) that can be supplied...2017 recently showed that by delivering CRISPR components as RNPs and delivering two halves of a repair...

...explaining my approach to conflict resolution, it’s important to recognize that there are different types of...or goal. Resolving these types of conflicts is important for the individuals involved, and also for the...That can mean a willingness to make yourself temporarily uncomfortable for the sake of a common good. ...with whom you strongly disagree, is a critical component of conflict resolution. Having empathy does not...transferring this skill to another job, it will be important to retain the stories and examples that have guided...

...this article, we’ll go through some of the most important considerations to make when planning and performing...same sample. For this strategy to work, it is important to ensure that you are using secondaries with ...primary, so we’ll start there. Some of the most important considerations are to make sure your secondary...publications or as a review on a company website. It is important to review the data and protocols available for...regardless of how “risky” they may be, it is always important to verify their performance in your hands and ...

... in advance Before validating your edit, it’s important to know a few things about your target cells and...goal is to generate a clonal cell line, so it’s important to verify that you won’t need to screen hundreds...method for knockouts is Tracking of Indels by Decomposition (TIDE) (Brinkman et al., NAR). This technique...trace files of WT and edited cell lines. Then decomposition analysis is performed yielding a graph of edits...assessment of genome editing by sequence trace decomposition. (2014) Nucleic Acids Research Volume 42, Issue...

...proteomics approach to monitor the composition of spatiotemporally defined protein complexes." Nature ...calcium uniporter (Martell et al., 2012). It is important to note that the biotin-phenol reagent can be ... of exogenous biotin allowing for more precise temporal control.  Find plasmids for TurboID and miniturbo...RapID consists of this mutant BirA* and an RNA component that contains an RNA motif of interest. Any proteins...

...screening out irrelevant candidates, and, more importantly, it’ll help you define salary costs. 4. Ascribe...BEFORE searching for a faculty position. This is important to ensure that you’re looking at institutes that...make these decisions in the following order of importance: 1. First make decisions based on experimental...and availability of the equipment is of utmost importance. Conducting a successful lab start up negotiation...resource budget, and know how to decide which components are the most critical to your research, negotiations...

...rates By Mike LacyGlutamate is one of the most important and abundant neurotransmitters, but it is challenging...targeting Glypicans By Ashley Waldron Glypicans are important modulators of cell signaling pathways during embryonic... domain structure. Glypicans are proteoglycans composed of a protein core, 2–4 glycosaminoglycans (GAG...swapping the homology arms for new targets. And, importantly, they found that moving the RNA untranslated ...fully understand how genes are regulated, it's important to know what proteins interact with each other...

...should I take to get to work?”; we often make important life decisions without much logic - even choosing... of work and communication before making this important decision. I’ll highlight some of these questions... if it is what is best for you. Some of these important questions will be easy to ask the lab head, and...rise of movements such as the Future of Research Symposium and #MeTooSTEM, academic bullies are hard to stop—they...information Overwork – including undue pressure, impossible deadlines, unnecessary disruptions Destabilization...

...1]. Like other transposons, piggyBac™ has two components, a transposon and a transposase. The piggyBac...useful for genetic engineering approaches. Most importantly for this technology, however, is that the transposase...should be tested in the cell line of interest. Importantly, by varying these ratios, you can either maximize...transposon sequences.  The use of PBx is particularly important to avoid any possible random re-integration from...lines, most likely due to DNA toxicity. More importantly, unlike Cre recombinase, the PBx is highly specific...

...review why validation is so important. Antibodies are a critical component of the researcher toolkit due...or plan your own validation experiments, it’s important to know what you are aiming to achieve. In general...as whole cell lysate or tissue sections. It is important to confirm that your antibody is not binding to...proteins bound to the target protein, so it’s important to understand if protein:protein interactions ...

...I have an important question: How much coffee does it take to fuel more than 30,000 neuroscientists over...needed to keep up with all the awesome talks, symposiums, poster sessions, exhibit booths, happy hour ...epileptic seizures in a mouse model. I sat in on a symposium discussing progress made in reprogramming astrocytes...scientific community. All of this speaks to the importance of scientific sharing to the neuroscience community...

...reason it is virtually impossible to determine specific protein turnover or temporal expression using standard...localization, or leading to protein aggregation. Another important parameter to consider is the contrast ratio- a...intensity, possible excitation lasers) it is also important to optimize your activation protocol for your ...photoactivatable fluorescent proteins Some of the most important applications for PA-FPs involve enhancing optical...