...address this need, the Khavari lab at Stanford created the RNA-protein interaction detection (RaPID) method...RaPID. Ramanathan et al. also tweaked BirA* to increase its rate of biotinylation. The improved enzyme...in neural progenitor cells. Knock-down of QKI decreased Zika viral RNA levels by 90%, suggesting that ...Hyperferritinemia-Cataract Syndrome (HHCS), a disease characterized by increased iron serum levels and early-onset cataracts, is...sequences and that loss of binding correlated with increased serum iron levels seen in HHCS patients. These...

...scaffold to enhance complex assembly, thereby decreasing the amount of background fluorescence from unbound...CRISPR to label multiple loci in distinct colors. To create a colorful Cas9 toolbox, Ma et al. turned to SpCas9...was fused to a different fluorescent protein to create three distinct colors. The specificity of these...distinct hairpin sequences produces white light. Created with BioRender.com. With this rainbow of colors...combination, for example, with a far-red FP, would increase the available colors to 15! They also envision...

...antibody and a ≥100-fold increase in signal over intrinsic fluorescence. Increased signal is likely due to...is a ~150 amino acid, single domain antibody. To create their RANbodies, Yamagata et al used previously... RANbodies are expressed in 293T cells and are secreted from cells due to their IgGκ signal peptide sequence...them. HRP RANbody staining results in a ≥10-fold increase in signal over staining with a primary/ labeled...

...Foundation, and Stanford University—that harnesses the creative power of sex and gender analysis for innovation... in research Governments, universities, and, increasingly, corporations in the US and Western Europe have...decades: “Fix the Numbers of Women” focuses on increasing the numbers of women participating in science...gender analysis and 2) provides case studies with concrete examples of how gender analysis leads to discovery... Environment—a project aimed at harnessing the creative power of sex and gender analysis for innovation...

...these demands? Are there other solutions, such as increasing numbers in Master’s level programs? What would...Maryland College Park, is a strong supporter of increasing the number of postdoctoral scientists at University...School. He has continually promoted the cause of increasing data collection about the scientific workforce... Boston 2015 where we hope to begin developing concrete solutions to the problems looming over the academic...School. In the lab, he uses synthetic biology to create tools to better understand human disease.  Outside...

...particularly given that this has precipitated increases to the NIH recommended pay scale – used by many...while this is a move in the right direction, the increase notably still does not meet the suggested postdoc...this area, keeping the pressure on for further increases will be challenging, but will be aided by the ... new strategies designed to cope with current increases. These two rulings along with progress over the...School. In the lab, he uses synthetic biology to create tools to better understand human disease.  Outside...

...grandsons that decreased their likelihood of death from cardiovascular disease but increased their risk of... DNA methylation patterns, either large increases or decreases. Hypermethylation of CpG islands in the... units of the FRET pair together and causes an increase in the FRET signal. Subsequent demethylation separates...

...needs to be compensated out of APC-Cy7. After increasing the compensation values for APC against APC-Cy7...to be shifted towards the x-axis, so you would increase the compensation value for APC. If the population... to be shifted away from the x-axis, you would decrease the compensation value.   Figure 1: Compensation...population needs to be shifted towards an axis, increase the compensation value, whereas if it the population...population needs to be shifted away from an axis, decrease the compensation value. If neither of these helps...

... aaRSs in these processes. To create the antibody toolbox, they screened antibody candidates using a synthetic...fourteen aminoacyl tRNA synthetases (aaRS) have been created, characterized, and validated by the Gräslund lab...cross-reactivity, antibody candidates underwent affinity screening to select for slow off-rate binders. The best ...pathways By Gabrielle Clouse Claudia Vickers’ lab has created a novel nanocompartment for transporting proteins...

...transfer plasmid at 30 °C instead of 37 °C and then screen for ITR recombination with a SmaI or XmaI restriction...duplicated regions like the ITRs. Restriction digest screening for ITR deletion is still necessary. To save time...mL of my AAV transfer plasmid cultures and then screen with SmaI digests. I freeze the bacterial pellet...how iodixanol purifications works, shows how to create the iodixanol gradient layers, and has some great...master iodixanol AAV purification. If you’re new to creating gradients, practice making the iodixanol layers...

...suppression of background by antibiotic selection. Screening of transformants by colony PCR confirmed the insertion...obtained if any single component was omitted. Screening for successful Golden Gate Assembly All Golden..., Table 1). These improvements (5- to 12-fold increases in the number of transformants while maintaining...to faster assembly workflows that require less screening to confirm the accuracy of assembly. The fidelity...test the limits of DNA assembly approaches with increasingly complex experimental conditions. The ability...

...amount of agarose/DMEM should be proportionately increased). For some cells, add a drop of serum to aid in...up, most will eventually grow within a week. We screen the wells in groups as they become confluent, taking...important to have a functional assay in place to screen your clones. Depending on the protein, we have ...SH-SY5Y cells, as these cells did not seem to form discrete colonies but constantly migrated away from each...either initially, or using secondary biological screens. For example, for cancer researchers, if growth...

... to its final destination. As an example, most secreted proteins are produced in the endoplasmid reticulum...golgi and then transported in vesicles via the secretory pathway to their subcellular or extracellular ... they are finally degraded (2, 3). Once you've created your fusion protein, it's important to validate... to make any fluorescent protein constructs you create in your own work available to the Addgene community...Edition. 3.  CureFFI.org. Cell Biology 04: The Secretory Pathway. 4. Dunn, Kenneth W., Malgorzata M. Kamocka...

...detectors for all fluorophores. This allows for the creation of much larger panels, as fluorophores that cannot...they don’t “wash out” the rest of the panel and create spreading error. Conversely, markers with low expression...should not be paired with these dyes, as this would create difficulty when trying to resolve those populations...signature upon antibody binding, leading to discrepancies during unmixing. Additionally, it is critical...spectral flow cytometry has the ability to generate incredible amounts of data from a single experiment. Although...

... deletions or homology-directed repair (HDR) to create precise genome edits. But have you heard of another...depositor Takashi Yamamoto’s lab has harnessed MMEJ to create a new method for CRISPR gene knock-in, termed PITCh...PITCh plasmids, GFP knock-in cell lines can be created in about a month and a half, without the need for...PITCh-gRNA, and the locus-specifc gRNA. The PITCh-gRNA creates two DSBs in the PITCh plasmid to release the cassette... alter expression of the gene it follows. For increased versatility, it would be advantageous to adapt...

... or the creation of bicistronic or multicistronic vectors. Unlike promoters which will create unique mRNA...resistance gene. These reporters allow you to easily screen or select for cells that are expressing your gene...interest at high levels. Unlike vectors that express screenable or selectable markers from a unique promoter,...now download Addgene's new eBook! Our goal was to create a one-stop reference guide for plasmids. We've ...

...researchers saw an 11-fold increase in mRNA levels after 2 hr and a 400-fold increase after 30 hr. The system...(CIB1) proteins. The goal of both groups was to create a system that would use light to turn on and off...domain to each of the Cas9 fragments, the authors created a photoactive Cas9 tool, as shown in the figure...binding and cleavage. This technique was applied to create photoswitchable spCas9, dCas9, and S. aureus Cas9...Gilgenast TG, Gong W, Beagan JA, Raj A, Phillips-Cremins JE (2019) LADL: light-activated dynamic looping...

...Komagataella phaffii (a.k.a  Pichia pastoris) because it secretes fewer of its own proteins, making the purification...production of Fab fragments where variable chains are secreted into the periplasmic space to be oxidized and ...incubation time to maintain cell viability and increase protein production (Schwarz et. al., 2020). Harvest...   Figure 1: The steps of rAb production. Created with biorender.com   Another method of harvesting...contaminating bands or a few strong ones, consider increasing the number of washes performed during your harvest...

... 2021). In fact, completion of Provost’s CURE increased students’ confidence in their research capabilities...projects have also been proposed as a mechanism for increasing diversity and inclusion in STEM disciplines (... Brownell, 2014; Duboue et al., 2022), as they increase student interest in scientific career paths while... Provost believes these networking experiences increase student enthusiasm, and research backs up that...undergraduate research experiences are a “CURE” that increases engagement by students and teachers. FEMS Microbiology...

...the templated DNA. Check out our blog post on increasing HDR efficiency to read more about optimizing ...pathways have been shown to be efficacious for increasing knock-in events (Yang et al., Int J Mol Sci)....culture media prior to and after Cas9 introduction increases HDR, just make sure to remove the inhibitors within...Early HR factors fused to Cas9 have been shown to increase HDR frequency by committing a break to performing...reading up on validating genome edits to find screening strategies for your experiment. Also learn about...