...spread the word as new protocols come out! References Noren, C. J., Anthony-Cahill, S. J., Griffith, M...incorporation of an unnatural amino acid in translation (Noren et al., 1989). In the late nineties, this was developed... But this focus on development, along with the inherent trickiness of using and optimizing GCE, led to...site-directed mutagenesis, many researchers new to GCE aren’t able to successfully complete their experiments...uncommon to find protocols that require reagents which aren’t broadly or easily available, so you may find yourself... they’d also love scientist interested in, or currently using, GCE to reach out to them and let them know...

... actually surprising to us that there aren’t a hundred different versions of this out there. Addgene: ...Addgene. For more details on the plasmids that are currently available along with detailed protocols, go to...and it’s already looking like it’s making a big difference. We can now look at many more transcription factors... this for tagging any protein in all kinds of different assays, not just transcription factors. The second...cells. It was made artificially. That means you aren’t getting any background or cross-reacting. We’ve... They could also switch from FLAG to GFP or a different epitope. Addgene: Do you have any additional tips...

...use temperature to denature and renature the dsDNA. Upon renaturation, you will generate DNA duplexes ...at how variable Cas9-expression was across the different cells (Figure 3).   Figure 2: Cas9 expression...publication. Pro-Tip: 1. Use multiple gRNAs to target different regions of your gene. I used existing BRAF gRNA...our website. There are a few reasons to target different regions of the gene with multiple gRNAs, but the...comparing the phenotypes of multiple gRNAs. Targeting different regions of the gene is also useful in case the...that would co-migrate on a gel) instead of two different sized digestion products (which could be easier...your way to making new scientific observations. References 1. Rago C, Vogelstein B, Bunz F. Genetic knockouts...

... probes, LDR can reliably detect single-base differences. The original creators of this technique visualized.... The structural/mechanistic reason for this differential is not known for certain, but may have to do...Sixteen separate pools were prepared, each with a different complement strand covering all 16 possible NN ...each possible pairing resulted in a product of a different length, with products identifiable and quantifiable... in the enzymology of DNA repair enzymes.     References 1. Shuman, Stewart, and Christopher D. Lima. ...superfamily of covalent nucleotidyltransferases." Current opinion in structural biology 14.6 (2004): 757-...PMC4737175. 22. Liu, Pingfang, Artur Burdzy, and Lawrence C. Sowers. "DNA ligases ensure fidelity by interrogating...

... away from the bench, I have worked for seven different organizations in a variety of science policy roles...opportunities. But even through all my roles and my current work as a AAAS Science and Technology Policy Fellow...cultural, and political. Science policy looks differently depending on how and where you pursue it. It ...Action Items:  Start a career journal during your current job and note what makes your heart sing, what makes...information may help you determine what parts of your current job you want to keep or change and can inform your...you do science policy? Just like there are many different types of science policy careers, there are many...NSPN maintains a great resource guide including conferences, societies, and outreach groups. Grab a virtual...

...perspective of the current. Although genome editing technologies have brought a renaissance in the field of...fiction, was also a trained biologist. This is apparent in his work The Island of Dr. Moreau (1896) where...In the 1960s, Har Gobind Khorana, Marshall W. Nirenberg, and Robert W. Holley cracked the genetic code...TALENs more specificity and a broader substrate preference than the ZFNs.   Genetic Engineering Era: Post...also transcended into the world of SciFi. This is apparent in the use of the word ‘CRISPR’ for anything associated...your research Browse plasmids for base editing References Anzalone AV, Randolph PB, Davis JR, Sousa AA,...pangenesis to the rediscovery of Mendel. Comptes Rendus de l’Académie des Sciences - Series III - Sciences...

...were also tested in different plasmid backbones that varied in size or carried different genes. In these ...promoters incorporate feedback loops such as the incoherent feedforward loop (iFFL). In general, iFFLs use...number. Moreover, gene copy number is affected differently by growth rates when a gene is expressed from...Segall-Shapiro et al. showed nearly no position-dependent differences in gene expression. On the other hand, slow growth...promoters can eliminate variability between the different components. When your pathway needs precise fine-tuning...levels, give these stabilized promoters a try. References 1. Segall-Shapiro, Thomas H., Eduardo D. Sontag...

... hand and use them to easily test different antibodies or different signaling molecules. Many labs find...proteins are represented in orange. Created with BioRender.com.    How do secondary antibodies work?  ...are readily available conjugated to a number of different signaling molecules. With the direct approach,...possible, or ensure that the primary antibodies are different species and/or isotypes. For highly multiplexed...has introduced new options for increasing signal strength.  For the time being, though, secondary antibodies...generated in goat against rabbit immunoglobulins. References and Resources More resources on Addgene’s blog...

... the retina using viral tools.   References and Resources References Pillay, S., et al. "An essential ...and can be specifically targeted to a number of different cell types even when systemically administered...capsids allow the virus to infect (or not infect) different cell types with some specificity. There are a ...will lose expression quicker than terminally differentiated, non-dividing cells.   Figure 2: ...Goz Ayturk with edits and updates from Addgenies Karen Guerin and Susanna Stroik. Didem Goz Ayturk is a...subretinal injections of neonatal mouse eyes at two different ages." Retinal Degenerative Diseases. Springer...

... What are the current numbers of trainees? Are there too few or too many at different levels? What does...generating too many trainees per PI” “Lack of awareness about how the system operates and functions” “...and outside of academic research (4). There are currently 17 sites around the USA undertaking BEST programs...a young scientist’s perspective, and the many different paths that biomedical graduates take in their ...policy. Follow David on Twitter @driglar.     References 1. Alberts, Bruce, et al. "Rescuing US biomedical...Henry, and Michael Roach. "Science PhD career preferences: levels, changes, and advisor encouragement."...

...wish to improve their current job situation. What can a coach do that is different from an adviser, boss... transition out of the academic sphere into a different meaningful career in science. Supervisors in the...speaking at research institutions and at scientific conferences, I see that few scientists are aware of the benefits...few hours of coaching time can make an enormous difference so don’t assume that this will cost a fortune...specializes in scientist coaching. Always ask for references and follow up with 3-4 to hear about those clients...also a very popular speaker and workshop leader. Lauren Celano of Propel Careers is not a professional ...

...than 300 different coactivator proteins individually contribute to cell regulation in a coherent manner.... one of the most successful targets for drugs currently available or being developed to treat a multitude...patterns of nuclear receptors across multiple different tissues in normal and disease states, their circadian...previously uncharacterized cellular roles. In the current phase of NURSA we are seeking to compliment these...our data mining tool, Transcriptomine. We are currently nearing the completion of an 18-month effort to...found in the Datasets and Reagents sections of the current NURSA website. NURSA and Addgene The great progress...

...life-threatening consequences. They result from defects in different steps in the glycan biosynthetic pathway and have...characterized by a predisposition to severe and recurrent infections along with a mild or exacerbated inflammatory...infection and other pathological conditions. References: Monticelli, M., Ferro, T., Jaeken, J., dos Reis...multi-subunit macromolecular complexes, but a serendipitous discovery led them to investigate the use of... discovered that the in vitro functions of 11 different Arg-mutant enzymes could be rescued by supplementation...diseases caused by Arg-mutant proteins, but there is currently no evidence of Arg-mutant protein rescue in living...from rare diseases arising from Arg mutations. References: Baldwin, Enoch, et al. "Generation of ligand...

... 1-4 different gRNAs. Viral envelope proteins To alter the cell tropism of Nanoblades, different viral...incubating Nanoblade particles with polybrene, a cationic polymer. Polybrene has long been used to increase...electroporation, and viral transduction; but these methods aren’t stealthy enough to gain access to some cells and...embryos, and animals. Nanoblades can also carry different types of CRISPR cargo including RNPs for indel...versatile and accessible genome editing tool.   References Mangeot, Philippe E., et al. "Genome editing ...

...job hunt, I quickly noticed one of the biggest differences between applying for an academic position versus... phone interview. I also transitioned between different lab groups without having a formal interview process...usually have a theme too, which is something not inherently captured by the STAR method. I like to think ... try and circumvent this, test out the video conferencing software you’ll be using in advance of the interview...interview, if possible. Otherwise, read the video conferencing instructions you received from HR carefully....completing each section of your talk. Some video conferencing software allows participants to “raise” their... wouldn’t have made it to the interview if you weren’t qualified for the position. So prepare for the ...

...their current promoters, as well as more promoters that target cells types missed by their current library... Clinical Ophthalmology Basel recently took a different approach to enhance specificity in retinal cells... synthetic promoters, the Roska lab used four different design strategies: Strategy #1 (ProA): Create... this library contained 226 AAVs, each with a different synthetic promoter driving expression of the same...was from a single class of cells  Of the four different promoter designs, promoters from strategy #4 had...ready-to-use AAV viral preps Browse all viral vectors References Juettner, J., Szabó, A., Gross-Scherf, B., Morikawa...

...never been so important.     References and resources References Biering SB,  Sarnik SA, Wang E, Zengel...for generating pseudovirus and those encoding different spike protein variants that can be incorporated...vaccinated individuals are at neutralizing the different SARS-CoV-2 spike variants. You can learn more ...head over to this blog post that highlights five different methods to use CRISPR to detect SARS-CoV-2. Other... including those from Andrea Pauli and Julius Brennecke. They developed a method that uses loop-mediated...Whiteman MB, McKay LGA, Lelis FJN, Habibi S, Cai Y, Rennick LJ, Duprex WP, McCarthy KR, Lavine CL, Zuo T, Lin...

...remember: Keep Ca+2lm and Ca+2rry on! References and Resources References Nasu Y, Shen Y, Kramer L, Campbell...sensors are commonly used calcium reporters with different kinetic parameters. There are three GCaMP types...and Localization The calcium concentrations in different subcellular compartments can vary greatly. Additionally...measurements are more susceptible to artifacts and pH interference, and comparison between experiments can be more...difficult due to variance in sensor expression if you aren’t working with a stably integrated biosensor. Ratiometric... also suitable for in vivo studies. There are different versions optimized for neuron-specific, astrocyte-specific...

...but timer activity can be calibrated to these different conditions, enabling the use of a given timer ...While Tsuboi et al developed mK-GO somewhat serendipitously while attempting to enhance mKO in other ways... of Neurog3 (a gene controlling pancreatic differentiation) in mouse embryos (Miyatsuka et al.) Distinguish... Let us know in the comments section below! References 1. Terskikh, Alexey, et al. ""Fluorescent timer...11090358. 2. Tsuboi, Takashi, et al. "Age-dependent preferential dense-core vesicle exocytosis in neuroendocrine...and Michael S. German. "Chronology of islet differentiation revealed by temporal cell labeling." Diabetes...programmable synthetic lineage-control network that differentiates human IPSCs into glucose-sensitive insulin-...

...have proposed the concurrent use of “orthogonal” Cas9 proteins derived from different bacterial species...variants—nuclease-competent or nuclease-null—in order to effect different perturbations. Consider, for instance, the challenge... both cutting and regulation of expression at different genetic loci within a single cell. One might imagine...expression (6). Altering the length of gRNAs targeting different sites of interest thus represents a straightforward...introduced together with truncated gRNAs targeting a different set of loci to induce simultaneous cutting and...DNA sequence or regulating gene expression.     References 1. Esvelt, Kevin M., et al. "Orthogonal Cas9 ...