...stranded RNA (ssRNA), while some exist as dsRNA. Still others use dsDNA to store their genetic information... made it into agriculture as species-specific pesticides and insecticides, an advancement over previous...sequence. All of the R-loops described above play distinct roles in promoting cellular processes and functions...While not perfect, these sensing systems work to distinguish between RNA of cellular vs. viral origin and ... each one acting on specific substrates and in distinct cell cycles. When RNA:DNA hybrids levels are allowed...appreciate double stranded isn’t just for DNA. Untwisting RNA is important for therapeutic delivery, viruses...

... gene expression. On the other hand, using a constitutive promoter in the same background resulted in ...over-represented compared to those further away. While constitutive promoters drove maximal gene expression near...during shifts in growth rate as compared to a constitutive promoter. To simulate the increase in plasmid...When they increased concentrations of IPTG, a constitutive promoter upstream of GFP increased expression...stabilized promoter while gene expression from the constitutive promoter did not. Similar levels of expression... al. placed the genes under the control of a constitutive promoter and then moved them into the genome...

... of interest in living cells. Challenges with existing RNA-protein interaction methods Not in a native...time point, but one minute of labeling with BASU still generated a >30-fold enrichment compared to E. coli...based proximity labeling methods, RaPID cannot distinguish between proteins that directly or indirectly ...sounds like a nifty technique, but what kinds of questions can it help answer? Ramanathan et al. outline ...of QKI decreased Zika viral RNA levels by 90%, suggesting that QKI is a host protein critical to Zika replication... study RNA-protein interactions. Do you have a question that RaPID could help answer? The RaPID plasmids...

...To LinkedIn or not to LinkedIn. That is the question. When presenting on building relationships (also...known as “networking”), one of the most common questions I receive is “Do I have to be on LinkedIn?” For...information available. When I am speaking at a new institution, for example, I will often see a whole bunch ...rule, but not that many). I’m asked a lot about requesting connections to complete strangers for informational...companies and organizations to see their news and job postings. 4) Use LinkedIn to pay it forward The best way... manager. This is considered intrusive and it’s still connecting with a stranger so is highly discouraged...

...participating in science and engineering. “Fix the Institutions” promotes gender equality in careers through...“Fix the Knowledge” or “gendered innovations” stimulates excellence in science and technology by integrating...study saved lives. Reduced use of estrogen plus progestin hormone therapy resulted in 76,000 fewer cases...the importance of analyzing sex, the National Institutes of Health (NIH) now requires that sex be included...Using cells from both sexes ensures that the diagnostics and therapeutic interventions you develop represent... work to be done! Researchers need to learn sophisticated methods of sex and gender analysis. Universities...

...regulation thats as sophisticated as a natural network. We could build a material as sophisticated as wood. To...Voigt explains it, his lab at the Massachusetts Institute of Technology has been “working on new experimental...to work with than regulatory proteins. There are still things we haven’t solved that stop it from being... carbons and oxygens. Nothing too much more sophisticated. There are also proteins, like antibodies. But...able to design new materials and chemicals as sophisticated as those that living cells and animals make....

...cycle once inside the host. While the virus can still enter (and thus infect) cells, this safety feature...Separation of retroviral packaging components onto distinct plasmids for safety.    Viral cargo-specific...cabinet and decontaminate after use. Virus-exposed plastics and other surfaces should be thoroughly decontaminated...decontaminated after use, typically with bleach (plastic) or ethanol (other surfaces). For AAVs, which are...should be added. After decontamination, single-use plastics and other handling tools should then be disposed... to reduce the risk of exposing yourself via needlestick or cut. Following all sharps procedures for your...

...past, we've been working with the Rare Genomics Institute to provide plasmid grants to researchers working...mechanisms behind the immune defects in CDG patients are still poorly studied and understood.  In line with this...complexes, but a serendipitous discovery led them to investigate the use of a common laboratory chemical, guanidine...Engineering the human disease alleles into cells for testing in the lab will be greatly facilitated by CRISPR...sites in T4 lysozyme by deficiency-creating substitutions." Journal of molecular biology 277.2 (1998):...significance (VUSs) that are frequently found in genetic testing for syndromes that include developmental delay,...

...other species, but these studies generally leave questions about gene function wide open. What if two seemingly...over time? Undergraduates can help to answer this question! The Pathways Over Time project The Pathways ... a framework for these undergraduate research questions. The idea is straightforward. Instructors first...have been using yeast models (3, 4) to answer questions about the evolution of the genes involved in methionine...experiments that are used to answer our research question, the plasmids are also used to teach students ...and instructor resources is available at our Investigations in Molecular Cell Biology Website. Many thanks...

...Nanoblades to Existing CRISPR Delivery Methods When compared head-to-head with existing CRISPR delivery...editing efficiency and off-target effects compare to existing technologies, and their use for HDR editing and...Table 1). Additionally, hiPSCs and mouse BM cells still behaved like iPSCs and BM cells after Nanoblade ...pluripotency marker expression and response to LPS stimulation, respectively. Primary cell type Gene targeted...to difficult to target cells. When compared to existing Cas9 delivery methods, Nanoblades have just as...

...An estimated 320,000 viruses can infect mammals. Even more abundant are the Earth’s estimated 1031 bacteriophages...instead to use the virus immediately upon harvesting. Interestingly, freeze-thawing host cells immediately... about the integrity of your virus, perform a diagnostic restriction enzyme digest and compare it to some...References 1. Anthony, Simon J., et al. "A strategy to estimate unknown viral diversity in mammals." MBio 4.5 ...

...deconstruct and understand this complexity using existing C. elegans mutants. For instance, we use a mutant...Minnesota has declared our makerspace an educational institution meaning that we can obtain known mutants at $7.00...makeup of the bacterial species in the gut of the domesticated housecat, and seeing how that makeup differs...cats with the goal of optimizing the health of domesticated cats and learning about feline microbiomes. ...pipeline. It’s possible that we’ll find something interesting and contribute it back to the kittybiome project...monitoring homemade microfluidic chips using the fantastic optics and fluidics platform it contains. Biotech...

...chance as entrepreneurs. To make things more interesting, we are working in a new technical discipline... cards, incorporation, laboratory lease, etc. I still flip to that page in my lab notebook from time to...worked weekends and evenings on the project, but still continued to generate new ideas as they popped into...doing makes sense. I can support that.” That was interesting. If people could be against GMOs as food but ... in the comments below. We’re happy to answer questions about this transition (so far) and what we’ve ...Biophysics and Biochemistry from the Illinois Institute of Technology in 2004, she accepted a commission...

...valuable laboratory tool are still an area of active research. New details are still being uncovered, and different...produce SuperFi-Cas9, a high-fidelity variant that still retains high cleavage efficiency (Bravo et al., ...et al., 2014). New discoveries on this topic are still being made! Conclusion Finally, we can summarize...their own advantages—and blind spots—to these questions. Each new structural insight also spurs the next...California, San Francisco, with support from National Institutes of Health R01-GM129325 and the Office of Cyber...Infrastructure and Computational Biology, National Institute of Allergy and Infectious Diseases. Resources...

...The vast majority of bacteria are undomesticated which limits the tools scientists can use to study them...CRISPRi machinery into diverse bacteria species. Existing protocols can transfer CRISPRi into a single bacterial...transfer Mobile-CRISPRi offers two advantages over existing bacterial CRISPRi methods. First, its modular ...Knockdown efficiency, as measured by knockdown of a constitutely expressed RFP reporter, was variable across ...Klebsiella pneumoniae, and P. aeruginosa (all opportunistic pathogens) were grown in presence of trimethoprim...Elizabeth A., and Pamela A. Silver. "Harnessing undomesticated life." Nature microbiology 4.2 (2019): 212....

...researchers have engineered alternative Cas9s binding distinct PAM sequences. Now, Hu et al., working in David...so far! Why do we need PAM flexibility? IDT estimates that there is an NGG PAM every 42 bases in the...edits or knock-ins. Editing efficiency decreases drastically once your cut site is more than 10 bp away from...frequency. Despite the progress we’ve made, there is still a need for and interest in enzymes with shorter ...editing; for known promiscuous gRNAs, xCas9 3.7 still showed a 4.2-9.4-fold improvement in off-target:...these early data are exciting, there’s a lot we still don’t know about xCas9. “I’m not 100% sure xCas9...

...after graduate school, at least not while you’re still in the thick of it. About three or four years into...ways to get a taste of a variety of careers while still working towards your degree. For me, experimenting...The article submitted was hilarious, full of interesting information, and received a lot of attention ... said, “Boy, I love science,” and I realized, I still do too! Beyond the outreach experience I gained ...back into lab the next day and remembered why I still love science. Keep building on your time in graduate...look back at a positive, fulfilling experience. I still use my network and graduate school resources daily...

...? Depositing lab: The name of the Principal Investigator (PI) in whose lab the construct was made. The...The depositor name links to a page listing all of all this PI’s plasmids that are available through Addgene... plasmid page.  The paper link leads to a page listing all the plasmids from this paper that are available...Recipient Scientist Acknowledgement (RSA) asks requesting scientists to cite the depositing lab whenever...about how the vector was assembled, and enables requesting labs to utilize and modify the plasmid in their...will be to your colleagues. If you ever have any questions about our plasmid pages, please don't hesitate...

...homology-directed repair Homology-directed repair (HDR) stimulated by double strand breaks has been widely used ...application, and the efficacy varies widely — it still almost always results in higher ratios of desired...cost of improved efficiency, although both enzymes still approach or exceed PEmax editing efficiencies for...obtain PEs small enough to deliver with AAVs but still efficient at long and complex edits. The team also...additive improvements. These variants are worth testing if you want to make your prime editing as efficient...this tail is degraded, the truncated pegRNA can still guide Cas9 and compete with full pegRNAs for access...engineered virus-like particles. However, there’s still a long way to go before we see prime editors in ...

...Chapter 1 asks “What is science?” That’s a very big question. Zimmer, a Professor of Chemistry at Connecticut...chapters in this section describe the challenges of trusting science in a “fake news and post-truth era,” the...would be complete without the inclusion of some interesting case studies, and Zimmer provides several. In...throughout this chapter. Now, these observations and suggestions are not so much inaccurate as already somewhat...period of time. Though climate change certainly still is and will continue to be a “critical consequence...current period of rapid flux, the chapter might still feel slightly dated by next week.   A stepping stone...