...response gene c-fos. For example, using Chr2 stimulation should result in robust increases in c-fos expression...after stimulation compared to controls without Chr2. c-Fos is a reliable and widely used molecular marker...proceed with your behavioral experiments. Comparison of c-fos amongst animals that receive different types of...neuronal activation, but it may still be possible to use c-fos expression to get the evidence you need. For example...activation in the novel context as determined by reduced c-fos expression, thus showing successful inhibition... as non-human primates [7, 8], zebrafish [9], and c. elegans [10]. If used properly, optogenetics is sure...but, in today’s post, we’ll discuss some of the important considerations you should think about when developing...

...future rare disease research funding opportunities. References 1. Jones, C., et al. "PP09. 1–2352: Systematic...disease are unclear. Dr. Di Schiavi plans on using C. elegans as a model to identify SMN1 interacting partners...effects of smn1 disruption. As part of his work in C. elegans, Dr. Di Schiavi will use Addgene plasmids...these genes, CRISPR to disrupt their function, and C. elegans overexpression plasmids to upregulate their... from rare diseases by connecting patients to important resources and promoting research for rare disease...Cancer Alliance Website.  We are proud to provide support to Drs Di Schiavi and Nabavi and will continue ...Blog Previous BeHeard Awardees Other Efforts to Support Rare Disease Research: IFOPA Donating Plasmids ...

...Y (T or C), X (any base), or C, respectively. gGBE, gTBE, gCBE Glycosylase-based G, T, or C base editors...transversion editors: CGBEs, or C → G Base Editors. CGBEs first convert a cytosine (C) to uracil (U), just like... a more specific A → C base editor (ACBE), they inserted the deaminase into the C-terminal domain of Cas9...pyrimidines (C ↔ T).   Base transversion Base change from purine (A or G) to pyrimidine (C or T) or vice-versa...type of base editor invented. CGBE C → G Base Editor Converts C → G by fusing a uracil DNA glycosylase..., or C, respectively, to be replaced with another base. References Chen, L., Hong, M., Luan, C., Gao... Zhang, D., Zhang, S., Li, C., Wu, J., Randolph, P. B., Sousa, A. A., Qu, C., Zhu, Y., Guan, Y., Wang,...

...transversions C → TG → A A → GT → C C → G                   C → AG → C                   G → T A → C                   ...Rosser, S. J., Bi, C., & Zhang, X. (2021). Glycosylase base editors enable C-to-A and C-to-G base changes...deaminase could also perform cytidine (C) deamination, converting the C base to uracil (U). This is the same...given the pithier name CGBE1, for “C → G Base Editor 1,” and had a C → G editing efficiency of up to 68%...Beyond UNG: Optimizing C → G editors Now that several labs had shown that preferential C → G editing was possible...pyrimidines (C ↔ T).   Base transversion  Base changes from purine (A or G) to pyrimidine (C or T) or vice-versa...type of base editor invented. CGBE C → G Base Editor Converts C → G; also called a glycosylase base ...

... 22nd International C. elegans meeting, I was again reminded of the extent that C. elegans researchers...and toolkits available at Addgene.  C. elegans open resources The C. elegans Neuronal Gene Expression Map...database specifically for C. elegans. This database provides detailed information about C. elegans 3’UTRs structures...UTRome The 3’UTRs of mRNA are important for post-transcriptional regulation in C. elegans. To study the role...The C. elegans community has always emphasized the need for open science and collaboration. The field...view the anatomy of worm layer by layer, the NeuroML C. elegans connectome, a model of the C.elegans neural...be used to simulate muscle tissue as it applies to C. elegans locomotion. To see the other resources included...

...for use in C. elegans. In 2017, the Korswagen lab developed Cas12a (Cpf1) for editing in C. elegans (Ebbing...editing from more sites of the C. elegans genome. Toolkits for editing in C. elegans The development of ...SapTrap-SEC kit for C. elegans genome engineering have also facilitated CRISPR editing in C. elegans. These...2018). To see more CRISPR tools for C. elegans, visit Addgene’s C. elegans CRISPR plasmids page. Future...Ruvkun for their seminal work on micro RNAs in C. elegans. The C. elegans work on small RNAs informed and drove... countless other systems, whereas import of CRISPR technology into C. elegans has been a transformative...possible in a wide range of organisms and cell lines. In C. elegans, one of the model organisms which I use to...

...bound to the C-terminal peptide. One of these C-terminal antibodies was Myc1-9E10 (aka anti-c-Myc [9E10]...sequences within the middle of the human c-Myc protein or the C-terminus, Evan, et al. isolated six different...that Myc1-9E10 and the other C-terminal antibodies only recognized the human c-Myc protein and did not recognize...experiments? Why Myc? When did the commonly used anti-c-Myc [9E10] antibody come into play? We’ll dive into...This protein had a highly specific antibody to the C-terminal five amino acids, meaning the tag could be...However, substance P has an amide group added to its C-terminus post-translationally, which turned out to.... As such, the peptide could only be added to the C-terminus of proteins of interest and proteins tagged...

...genome engineering methods for C. elegans were released, signaling a new era in C. elegans research. Homology...of custom genomic modifications, is very robust in C. elegans, and the methods for HDR-mediated modification...the University of North Carolina has made CRISPR in C. elegans even easier - now, one can generate a fluorescent...-based gene tagging approach Multiple features of C. elegans have made CRISPR/Cas9 so successful in this...previously used a transgene selection strategy for their C. elegans/CRISPR experiments, but this strategy required... make it possible fluorescently label proteins in C. elegans, they inserted the SEC into a synthetic intron...system is both simple to use and highly efficient for C. elegans genome modification. Figure 2: Schematic...

...the C-terminal region of the human proto-oncogene c-Myc. Since the generation of the first anti-c-Myc ... on the Anti-c-Myc [9E10] page for more details. Image credit: Addgene.   Find anti-c-Myc antibody...potassium channelrhodopsins Myc-tag antibody anti-c-Myc [9E10] now available! Improved voltage indicator...and tdTomato expressed layer 2/3 neurons in mouse. C) Action potential frequencies with (magenta) and without...10.1101/2021.09.17.460684.)   Myc-tag antibody anti-c-Myc [9E10] now available! by: Ashley Waldron The majority...pulled up over 13,000 plasmids! Our recombinant anti-c-Myc [9E10] (isotype IgG2a) antibody is currently recommended...Isolation of monoclonal antibodies specific for human c-myc proto-oncogene product. Mol Cell Biol. Dec;5(12...

...fourth-generation base editors, BE4, reduce the undesired C→G or C→A conversions that can happen with BE3s. These...Compared to BE3, BE4 offers a 2.3 fold decrease in C→G and C→A products as well as a 2.3 fold decrease in indel... Hong, M., Zhang, D., Luan, C., Zhang, S., Wu, H., Gao, H., Bai, S., Li, C., Ding, R., Xue, N., Lei, Z... W., Levy, J. M., Yeh, W.-H., Zheng, C., Newby, G. A., Wilson, C., Bhaumik, M., Shubina-Oleinik, O., Holt...) and adenine base editors (ABEs). CBEs mediate a C to T change (or a G to A change on the opposite strand...strand). ABEs make an A to G change (or a T to C change on the opposite strand). Both are base transition...excision repair, which would change the U back to a C base. To increase base editing efficiency, Komor and...

... and C-C’) that give the feature its characteristic T-shape. The arrangement of the B-B’ and C-C’ palindromic... Tian, W., Liu, C., Lian, Z., Dong, X., & Wu, X. (2017). Deletion of the B-B’ and C-C’ regions of inverted...contains three palindromic sequences (A-A’, B-B’, and C-C’ regions) and forms a T-shaped structure. The Rep... Conatser, L. M., Lue, V. M., Dobbins, A. L., Li, C., Hirsch, M. L., & Samulski, R. J. (2020). Adeno-Associated...AAV plasmids contain several important elements, but among the most important are the inverted terminal ...Adeno-associated viruses (AAV) are an important biological tool that allow scientists to express a gene...modifications and types  Since ITRs are one of the most important elements of AAV and rAAV vectors, modifications...

... with 40–60% G/C content are most likely to be successful, although sequences with G/C content outside...of the 3′ extension of the pegRNA should not be C. A C base is speculated to base pair with G81 of the..., J., Oyler-Castrillo, P., Ravisankar, P., Ward, C. C., Levesque, S., Jing, Y., Simpson, D., Zhao, A.,...Figure 3: pegRNAs whose 3′ extension begins with a C base can cause unwanted base pairing (left), while... Koblan, L. W., Levy, J. M., Chen, P. J., Wilson, C., Newby, G. A., Raguram, A., & Liu, D. R. (2019). ... Knipping, F., Ravisankar, P., Chen, P.-F., Chen, C., Nelson, J. W., Newby, G. A., Sahin, M., Osborn, ..., Anzalone, A. V., An, M., Newby, G. A., Chen, J. C., Hsu, A., & Liu, D. R. (2022). Engineered pegRNAs...

...editing toolkit to allow C to U edits using their RESCUE (RNA Editing for Specific C-to-U Exchange) system...RESCUE-S) offered ~76% on-target C-to-U editing while reducing off-target C-to-U edits by ~45% and off-target...lab published a paper on an RNA editor that allows C to U edits (RESCUE). We’ll walk through how these ...adjusted the structure of the guide RNA by introducing a C opposite the target A to be edited. This change specifies...residues that contact the RNA substrate resulted in 15% C-to-U editing activity. They then performed sixteen...cytidine deaminase activity. While RESCUE can perform C-to-U edits, the authors note that it retains its adenine... that RESCUE is most active in a 30 nt guide when C or U base-flips are present across from the target...

...in fact, permeable to CO2 at low temperatures (-80°C) and once in the tube, the CO2 can alter the pH of...pH indicator phenol red. We froze samples in a -80°C freezer and then moved them to a closed dry ice storage...samples to controls that were only stored in a -80°C freezer and not exposed to CO2/dry ice. All samples...then tested whether returning the tubes to the -80°C freezer after storage on dry ice allowed the CO2 to... (Fig.4). To test this, we froze the tubes at -80°C and then moved them to closed storage on dry ice for...for 48 hours. We then moved the tubes back to a -80°C freezer for various recovery periods between 0 and... samples to controls that were only stored at -80°C. We then thawed the tubes after the indicated incubation...

...fluorescent protein is at the C-terminus of the POI or that the POI is at the C-terminus of the fluorescent...dimerization (Shaner et al., 2008 - supplementary figure C). However, these assays are qualitative. The Organized...structural information available, it’s clear that the C-terminus of fluorescent proteins derived from Aequorea...biosensors (yellow cameleon 3.60, EPAC, and Galphai), the C-terminus and N-terminus of the donor fluorescent protein...the FRET efficiency. Creating fusions at the N- and C- termini of your protein of interest The most straightforward...straightforward fusion protein is made by connecting the N- or C-terminus of the fluorescent protein and protein of... the sequence, people often use terminology like “C-terminal fusion,” but I find this very confusing because...

...expressed to identify successfully transfected cells. C-to-T editing by the APOBEC-Cas9 editosome would restore...codon that is converted to a ‘TAC’ or ‘TAT’ after a C-to-T base editing event. This edit changes the histidine... ATG codon from an ACG codon using a base-editor (C>T conversion) they could induce the translation of...Liu DR (2017) Programmable base editing of A•T to G•C in genomic DNA without DNA cleavage. Nature 551:464... Figure 4: Reporters in Gene On include a mScarlet reporter, luciferase reporter, and neomycin resistance... editing. eGFP reporters for single base editing by APOBEC-Cas9 Though the ACE reporter quantifies APOBEC-Cas9...DSB in their previous reporter, the Harris Lab created a panel of eGFP reporters to quantify the on-target...

... Lucia, C., Federico, P.-B., & Alejandra, G. C. (2020). An ultrasensitive, rapid, and portable coronavirus...Time: ~40 minutes Equipment required: 37°C heat block and 62°C heat block Approach: Viral RNA is extracted...range Time: ~1 hour Equipment required: 37°C water bath and 42°C water bath Approach: RNA is extracted from...Valois, E., Solley, S. C., Braig, F., Lach, R. S., Baxter, N. J., Kosik, K. S., Arias, C., Acosta-Alvear, D...incubation  Time: ~90 minutes Equipment needed: 37°C heat block, LED blue light illuminator or UV light... in a range of temperatures varying between 10-50°C. Visit Addgene's COVID-19 Plasmids and Resources ...Sotomayor-Gonzalez A, Zorn K, Gopez A, Hsu E, Gu W, Miller S, Pan C-Y, Guevara H, Wadford DA, Chen JS, Chiu CY (2020) ...

...j.cell.2021.09.018 Choi, J., Chen, W., Suiter, C. C., Lee, C., Chardon, F. M., Yang, W., Leith, A., Daza,..., J., Oyler-Castrillo, P., Ravisankar, P., Ward, C. C., Levesque, S., Jing, Y., Simpson, D., Zhao, A.,...Leukemia Virus (M-MLV) reverse transcriptase to the C-terminus of Cas9 H840A nickase (PE1). But innovations...pegRNAs (Yan et al., 2024). The team fused La to the C-terminal end of PEmax to produce PE7. PE7 significantly...expression ~ ~ ~ ~ ✓   ✓ SV40 and c-Myc nuclear localization sequences Improve translocation...References Anzalone, A. V., Gao, X. D., Podracky, C. J., Nelson, A. T., Koblan, L. W., Raguram, A., Levy... Koblan, L. W., Levy, J. M., Chen, P. J., Wilson, C., Newby, G. A., Raguram, A., & Liu, D. R. (2019). ...

...institute. Set your -80 °C freezer to -70 °C and teach other labs to do the same. -70 °C is cold enough to keep...years. In fact, some scientists speculate the -80 °C setting might have been a marketing campaign! MIT ...biological samples that are stored long term at -70 °C. My Green Lab also has resources and even a freezer... list some easy but effective habits you can incorporate to reduce your lab consumption and increase your...in your daily habits can feel ineffective or unimportant. With so many big contributors to the sustainability...

...Tobey, I. G., Moncada-Reid, C., Eid, F.-E., Walkey, C. J., Ljungberg, M. C., Lagor, W. R., Heaney, J. ...Ravindra Kumar, S., Adams, C. D., Yang, D., Wang, T., Wolfe, D. A., Arokiaraj, C. M., Ngo, V., Campos, L....Gong, Y., Ng, C., Nammour, J., Maiz, D., Dujardin, S., Ramirez, S. H., Hudry, E., & Maguire, C. A. (2019)... Poterba, T., Boutros, C. L., Balazs, A. B., Daneman, R., Bloom, J. M., Seed, C., & Deverman, B. E. (2019...expression control. (c) Viral delivery methods include focused ultrasound (FUS), intracorporeal (IC), intracerebroventricular...to the brain. The AAV (a) capsid, (b) cargo, and (c) delivery method determine cell type- or region-specific...Challis, R.C., Ravindra Kumar, S., Chan, K.Y., Challis, C., Beadle, K., Jang, M.J., Kim, H.M., Rajendran, P....