...fields where communication and writing are key skills. Other fields, often highly technical or skills-based...proactively address questions about your application. These could be logistical, like briefly explaining a gap ...excellent communication skills” try giving an example, like “I have extensive communication experience...usually yes! A good cover letter won’t hurt your application, but it can certainly help you stand out.  In...consider cover letters an important part of the application. Even within an organization, there can be variance...good cover letter!) As with any part of the job application process, it's a good idea to talk to people in...work on [phenomenon X], I read several of your publications on [specific topic]. I am extremely interested...

...revolves around synthetic humans called replicants which are genetically engineered by a powerful corporation...fused a catalytically impaired Cas9 protein to a DNA-editing enzyme to generate the prototypical base editor...genetic code of living organisms and its societal implications even before the discovery of genes themselves...explores the concepts of animal testing and modifications. The premise of the novel involves Dr. Moreau...technology led to the development of many important medical and agricultural products. But it also made SciFi...2018) were highly successful in the box office, indicating the insatiable appetite of the moviegoers for...a theme park.  Released in 1997, GATTACA is an American SciFi film starring Ethan Hawke, Uma Thurman and...

...and GFP-edited amplification products (BRAF-GFP duplex) and from GFP-edited amplification products (GFP-GFP... wanted to inactivate the gene BRAF (a kinase implicated in several human cancers) in A549 cells (a human...via transduction with Cas9 lentiviral vector, lentiCas9-Blast (52962-LV, Figure 1). Some cells were then...Cas9 expression in A549 cells transduced with lentiCas9-blast. Anti-Cas9 Western blotting was performed...transduced with Cas9 expressing lentivirus at the indicated multiplicity of infection (MOI). Anti-vinculin...lines generated from A549 cells transduced with lentiCas9-blast. A549 cells were transduced (MOI = 37) and...need to use FACS (like they did in the original publication). I thought I could see GFP expression with a...

...prior publications to go on, a submitted manuscript could only have been viewed as a significant improvement...causes malaria). I attribute their success to technical expertise, thoughtful single guide RNA (sgRNA)...the piñata builders - the reviewers. After some physical exertion on the part of the party goers, the piñata... solutions that facilitate most experimental applications in CRISPR engineering. When compared to that...easy to fault the lack of features in a single application. The question is then, should we use all tools...those engaging in CRISPR design, but it is a significant hurdle made worse by an ever increasing marketplace...the other hand some software tools make use of empirically derived and/or prior knowledge to inform on the...

...natural redundancy of biological pathways;  to observe a phenotype, the modification of multiple genes is...recognition and cleavage. This sequence flexibility indicates that gRNAs could be incorporated into these constructs...the 5’ end. PTG construct schematics. Scissors indicate cleavage sites. After confirming gRNA cleavage...) How many loci can Cas9 modify? To test the application of this system to gRNA multiplexing, Xie et al...deletion with 4-20% frequency at the four loci, indicating that Cas9 can be guided to at least 8 distinct...multiplexing further illustrates the potential of PTGs. Applications of PTGs The major advantage of the PTG system.... Though the plasmids made by Xie et. al are specifically designed for plant expression, PTGs will likely...

...Stanford University. Sex and gender are critical components of biological research that are often forgotten ...provide a table indicating the sex of the most commonly used cell lines appearing in American Journal of Physiology-Cell...the past decade. For more practical help, there are numerous methodological articles to consider—we have... that of human clinical trials. And, as Janine Clayton points out, late-stage clinical studies that draw...effects of using all female cells and tissues, specifically in stem cell therapies. Let’s go back to why... disease being treated, and other hormonal, immunological, and environmental factors. How to include sex... (NIH) now requires that sex be included as a biological variable in all NIH-funded research. Janine Clayton...

...normalization and loading controls needed for relative quantification of a western blot — and why you might want ...accurately compare different lanes? In relative quantification, the POI bands are normalized to the loading...other errors in experimental design and gel quantification. Figure 3: In a perfect world, protein...On this gel, the protein of interest band is significantly lighter in “condition 1” than “condition 2.”...and it’s also problematic for computational quantification. Outside the linear detection range, band intensity...common error associated with Western blotting quantification” (Gilda & Gomes, 2013). To address this source... Gilda & Gomes, 2013. Figure 7: Semi-quantification of liver lysate using different loading controls...

...proteins, such as receptors, or post-translational modifications on those proteins, like glycosylation patterns...Depending on the research question and downstream application, one or several of those techniques can be used...populations; for example, CD8+ T (“T killer”) cells. Typically, you would label the CD8 co-receptor protein on...your flow analysis. However, certain downstream applications, like an in vivo adoptive transfer of those ...cell membrane but inside the cell. Due to their chemical nature, antibodies cannot penetrate the cell membrane...intracellular labeling with antibodies requires chemically pre-treating the cells to allow for entry of ...but preserving the cellular state (think of mummification). Permeabilization, as the name suggests, perforates...

...current CRISPR research focuses on its potential applications for human medicine (Waltz, 2016), the potential...engineering, on the other hand, allows for targeted modification of known or suspected genes that regulate a ...commonly used model organisms like Arabidopsis and Medicago truncatula and several crop species including ...engineering using CRISPR, highlight the specific modifications to CRISPR machinery that allow for the use of...CRISPR principles). However, plant-specific modifications to commonly used CRISPR plasmids are necessary...selected the correct CRISPR components for your application, it is time to deliver these components to your...detergent, Polyethylene Glycol (PEG), although the application of this approach is limited to protoplast cells...

...exclude duplicate events, and the Live/Dead gate to focus your analysis on what’s biologically active....exclude all duplicate events (two cells stuck together) from your single cells, as duplicates can’t be reliably..., you’ll first need to learn a little bit of theoretical background on the different parameters that your...and W describe the shape of a histogram, which graphically represents the time and intensity of the cell... fully depend on your individual experiment. Practical example: lymphocytes in a mouse tumor sample You... You made it! So much about the theoretical background. Let’s dive into the praxis of gating strategies...show up in a straight line one the plot, while duplicates deviate from that pattern. A.                                                                                                         ...

...strategies Publication System nickname Photoactivatable moiety Cas9 variatn CRISPR protein application Split...improvements to CRISPR technology since its initial applications for genome engineering in 2012.  Many of these...temporal and spatial control. Optogenetics uses genetically encoded tools, such as microbial opsins, to control...enzymatic activity. Photoactivatable genome modifications by NHEJ and HDR using split systems Later in...mutations via NHEJ (frequency of 20.5%) and induced modifications by HDR (frequency of 7.2%). The authors additionally...C714, generating paCas9-2. This change did not significantly alter the system’s efficiency at generating ...particular interest since blue light has limited applications in vivo. UV and blue light is readily absorbed...

... do anything. Viruses are complicated and they perform complex biological tasks, both of which are factors...C virus RNA quantification as measured with branched-DNA technology." Journal of clinical microbiology...factors in the technical challenges they can present. Many researchers find themselves plagued by troubleshooting...hail Marys that have helped me troubleshoot some technical obstacles when working with virus: 1. Viral vectors...vector may undergo genomic rearrangements during replication. To avoid genomic rearrangements, try amplifying...Different titering methods can reveal differences in physical versus functional titer. If you have low titers...cells, replace the medium with half the volume typically used (e.g., use 5 mL medium on a 10 cm plate)....

...the OpenWorm website. microPublication microPublication is a unique publication for the worm community...into larger publications and would otherwise be lost in lab notebooks forever. microPublications are peer-reviewed...aims and approach of CeNGEN, check out their publication.   3’UTRome The 3’UTRs of mRNA are important ...Biodesign institute developed a 3’UTR database specifically for C. elegans. This database provides detailed...normally one figure and can be negative and/or even replicated studies to show reproducible results. This a ...referenced. In addition the data published in microPublications is curated and added to other databases including...including wormbase and flybase. microPublication is run and maintained by members of the worm community...

...experiments. However, the technical demands of using these wireless devices are significantly higher than standard...optic cable is threaded through—and fixed to—an optical insulator called a ferrule (below). The fiber optic...portion of optogenetics (though it does require a significant amount of equipment, most of which is relatively...There are several important considerations with fabrication of the fiber optics needed for optogenetics. ...point; the fiber optics need to be polished to significant smoothness to prevent drastic loss of efficiency...maintained during the entire protocol, otherwise optical stimulation will be erratic, opsin activation/inactivation...stimulation, the rate of movement of the animal significantly increases. Before you get your experiment started...

...Rep78/68 play a key role in AAV genome replication. Replication initiates from the AAV genome ITRs on ...Srivastava A (2019) Capsid Modifications for Targeting and Improving the Efficacy of AAV Vectors. Molecular...contains the proteins needed for the virus to replicate,  and the transfer plasmid which contains the ...aka the RepCap plasmid encodes two genes: the replication (rep) gene and the capsid (cap) gene. In wild-type...its products. Created with BioRender.com   Replication (rep) gene The rep gene encodes four proteins...proteins that are required for viral genome replication and packaging: Rep78, Rep68, Rep52, and Rep40. Rep gene... domains, all four Rep proteins share a common helicase and ATPase domains. To function, the Rep proteins...

...origin of replication (which controls the host range and copy number of the plasmid) and typically include...Description Origin of Replication (ORI) DNA sequence which allows initiation of replication within a plasmid...point for PCR amplification or sequencing. Primers can be exploited for sequence verification of plasmids...plasmids are small circular pieces of DNA that replicate independently from the host's chromosomal DNA....Minimally, lab-created plasmids have an origin of replication, selection marker, and cloning site. The ease... plasmids and the ability of plasmids to self-replicate within a cell make them attractive tools for the...plasmid by recruiting replication machinery proteins Antibiotic Resistance Gene Allows for selection of plasmid-containing...

...   Figure 3: Each publication featured in BioGRID ORCS has a dedicated summary page with PubMed...result interpretation. The analytical method for score assignment and significance thresholds used by the ...favorite gene was described in a CRISPR screen publication. You see this mentioned in the results section...new freely available resource developed by the Biological General Repository for Interaction Datasets (... to allow facile queries of published data by biomedical researchers and ready access to stably archived...manually curated and annotated from more than 45 publications, beginning with the seminal research articles...CRISPR screen results available to the entire biomedical research community, but also allows cross-interrogation...

...their undomesticated counterparts) - not exactly species that jump to mind when we think “biomedical research...ways these animals have impacted humans through biomedical research. The Camelid family is composed of ...research". The family entered the biomedical field’s spotlight after researchers discovered heavy-chain...to sdAbs becoming valuable tools for diverse applications. For example, RANbodies can be used as an alternative...chromobodies,” which are single-domain antibodies genetically fused to fluorescent proteins and used to visualize...therapeutic was approved in 2018, with many others in clinical trials (Jin, et al. 2023). In addition to therapeutics... excitement around using sdAbs in diagnostic applications. For example, fluorescently and radioactively...

...May 22, 2019 Smart Servier Medical Art This site has 3,000 free medical images organized into four main...software that let me create graphics that better communicated the science in my blog posts. This post highlights...free tools which will hopefully also help you communicate your science, whether it’s in presentations, ...number of drawing tools, so it’s harder to draw intricate figures, like a brain or a mouse. Vectr Vectr ... over 20,000 icons is designed by professional medical illustrators with input from life scientists. It...without the watermark, which are better suited for publications, if you sign up for a paid account.  I’m proud... anatomy and the human body, cellular biology, medical specialities, and general items. Individual images...

... tactics of prokaryotes to deliver the technological biological blockbuster known as the CRISPR (clustered...genes, which code for polymerases, nucleases, and helicases, were also disrupted to elucidate their role in...destroy the phage upon re-exposure (2, 3). More specifically, specialized Cas proteins snip the foreign DNA...distinct types and subtypes using polythetic classification based on evolutionary similarity (5). Type ...2013 (7, 8). Nine months later, more than 1500 publications describe work to improve the tool’s specificity...various species and describe an assortment of applications. Despite the ubiquity of CRISPR/Cas toolkit,...programmable platform well-suited for various innovative biological and translative technologies. Addgene has empowered...