...experiments or even negatively impact cells (see Figure 1). When behaving properly, FPs enable researchers ... disulfide bonds in the secretory pathway (Figure 1) [4,6–8]. Additionally, FPs that encode N-glycosylation... sarcoma-associated herpesvirus.       References 1. Snapp, Erik. "Design and use of fluorescent fusion...genes: deletion mapping functional domains of Drosophila hsp 70." The EMBO journal 3.13 (1984): 3087. ...

...another great year of sharing!   References 1. Shalem, Ophir, et al. "Genome-scale CRISPR-Cas9 knockout...Central PMCID: PMC4089965. 2. Sanjana, Neville E., Ophir Shalem, and Feng Zhang. "Improved vectors and genome-wide...

...sequences were initially discovered in E. coli in 1987 (1), the concept that these clustered repeat sequences...neutralize CRISPR/Cas. Earlier this year, expression of 1 of 5 anti-CRISPR genes were found to inactivate the...light until 2007. Initial experiments exposed S. thermophilus to predatory phages to test if the exogenous...

...has error rates that range from roughly 1 per 500bp to roughly 1 per 10 million bp depending on the polymerase...analyzing the gene’s function. The process is shown graphically in the following figure, in which we add EcoRI...

...vectors and include a Kan, SpHIS5, or CaURA3 selection marker. Lee et al.(1) assessed many of these fluorescent... Shah NG, McKenzie A 3rd, Steever AB, Wach A, Philippsen P, Pringle JR. Yeast. 1998 Jul;14(10):943-51....3rd, Demarini DJ, Shah NG, Wach A, Brachat A, Philippsen P, Pringle JR. Yeast. 1998 Jul;14(10):953-61....

...construct for crystallographic and biophysical studies of protein degraders. Nat Commun., 15(1), 8885. https...and is recommended for use in western blots (Figure 1). If you prefer to produce your own antibody or want...are also available from Addgene!   Figure 1: Experimental validation for Anti-V5 [IPI-SV5-Pk1]...

...guide RNA (sgRNA), provided it meets two conditions:  1) The sequence is unique compared to the rest of the...bystander bases” if you choose this system.  Fig. 1: Cytidine deamination takes place on the free strand...temperature-controlled genome editing. Nat. Commun. 2017 81 8, 1–9 (2017).Walton, R. T., Christie, K. A., Whittaker...(but not limited to) bacteria, yeast, plants, drosophila, worms and mammals (Browse plasmids by model ...

...animal will repeatedly nose-poke at the active hole [1, 2].    Place preference/place aversion also models... can be explained primarily in two different ways 1) your hypothesis is wrong or 2) there is some problem...follow him on Twitter @derekpsimonphd.    References 1. Stuber, Garret D., et al. "Excitatory transmission...technology is improving and may one day reach the sophistication of mouse transgenics. Everything that has been...

...sequence on either side of the desired mutation (usually 1-3 mismatched bases) is sufficient for your primers...researcher at Cornell University.       References 1. Breslauer, Kenneth J., et al. "Predicting DNA duplex... PCR followed by Restriction Fragment Length Polymorphism (RFLP) analysis (right). Introduction of restriction...of that particular site with fragment length polymorphism (RFLP) analysis. In this process, if your mutation...

...total dose between 1 x 1011 and 5 x 1011 vg per animal (i.e., 10 and 50 µL of vector at 1 x 1013 vg/mL titer... and 1 x 1012 vg*. Most systemic capsids are easy to produce in high titer, but higher doses (>1 x 1012...before using it in an application.     Figure 1: AAV toolkit for targeted gene delivery to the brain...decision tree for selecting a capsid, while Table 1 gives a brief overview of published descriptions of...Capsids page to learn more about that collection. Table 1: Systemic capsid plasmids available from Addgene ...AAV9. At lower viral titers than AAV-PHP.B (e.g., 1 x 1011 vg/mouse), AAV-PHP.eB can transduce a greater...Diverse Brain Cells. Annual Review of Neuroscience, 45(1), 447–469. https://doi.org/10.1146/annurev-neuro-111020...

...Resist the urge to say everything I allow 1 slide for every 1 minute of talk length, and then I cut a few...NatureJobs: Interviews: The All-Important Job Talk. 1. Watch other people give great talks We are not talking...Like graphing data, choosing controls, or mixing clear solutions—public speaking is skill that any scientist...

...prompted us to create Crowdfight.   Figure 1: Crowdfight is a community of scientists who believe...collaborator for a specific task, Crowdfight will: 1) Find an expert with the right skill; 2) Confirm the...volunteers helped with things like translations, bibliographic searches, etc.     Expanding collaborations...

...for locating and containing escaped flies. Nature 4:1-22. https://doi.org/4.01.2022/hafd...FLIS and Flies  Drosophila, more commonly known as fruit flies, have been an important animal model for...

...Hsp70, the heat shock releases heat shock factor 1 (HSF-1), which subsequently binds to heat shock elements...genome engineering in species like C. elegans and Drosophila. Light inducible promoters Light is another way...

...deletions, and all possible base-to-base changes.   1. Plasmids 101: Transformation, Transduction, Bacterial...Scientific Graphics by Beth Kenkel Find a round-up of tools you can use for making scientific graphics. PS:...

...Introduction to Biosensors. Essays Biochem. 2016 Jun 30;60(1):1-8. DOI: 10.1042/EBC20150001 Leopold, A. V., Shcherbakova...of the biosensors used in research are very sophisticated, but at their core they can all be boiled down...

...modified slightly with either a 1-7 base poly(U)tail in tRNA-gRNA constructs or a 1-4 base tail from the second...three strategies described in this post in the graphic below - including some simple comparisons. The...

...light-sensitive LOV2 domain of Avena Sativa phototropin 1 (AsLOV2). In the dark, this peptide is caged by the...iLID nano and 800nM to 47µM for iLID micro; Figure 1). With this great affinity range, these LIDs have ...Figure 2). They are all available now at Addgene. 1 Lungu et al., Chem Biol. 2012 Apr 20;19(4):507-17....Guntas et al., Proc Natl Acad Sci USA 2015 Jan 6; 112(1):112-7. doi: 10.1073/pnas. CRISPR-Cas9 optogenetic... al., BMC Molecular Biology. 2013 August 20th; 14(1). TRICK: A method for visualizing the first round...for a variety of experimental purposes including (1) single or multiple, constitutive or inducible, gene...the years. The authors optimized these systems by: 1) decreasing the number of plasmids required for each...

...post about multiplexing with Cpf1.   Figure 1: Multiplexing allows researchers to express multiple...be used in a wide range of bacteria. References 1. Maddalo, Danilo, et al. “In vivo engineering of oncogenic...other model systems, including E. coli, plants, Drosophila, and zebrafish!   Why use multiplexed gRNAs?...restriction enzyme BsaI. If you’re working in Drosophila, a two-gRNA expressing plasmid is available from...supplied on a separate plasmid. Multiplexing in Drosophila Bullock Lab Multiplex Plasmid: Two gRNAs can ...or SLIC cloning. gRNAs are expressed from two Drosophila U6 promoters. Cas9 must be supplied on a separate...efficient germline and somatic genome engineering in Drosophila.” Proceedings of the National Academy of Sciences...

...healthcare system was $35.2 billion (or $140 per $1 invested in the trial). The goal of gendered innovations...is the goal of Gendered Innovations. This project: 1) develops state-of-the-art methods of sex and gender... work to be done! Researchers need to learn sophisticated methods of sex and gender analysis. Universities...