...immediate early gene expression. Genes Dev. 1988 Jun;2(6):718-29. doi: 10.1101/gad.2.6.718. PMID: 2843425...Targeted Gene Expression in Drosophila.” Genetics 186(2) (2010): 735-55. PubMed PMID: 20697123. PubMed Central... in Saccharomyces cerevisiae.” FEMS Yeast Res. 12(2) (2012): 197-214. PubMed PMID: 22129153. Additional...system to GAL4/UAS that functions in essentially the same manner, with LexA binding to the lexA operator (...desirable to control or look at multiple genes at the same time.  The 3 binary systems listed above are orthogonal...

...label) your sample (i.e. cultured cells or tissues).  Step 2: attach anchors to your sample that will link...so much. Created with biorender.com Steps 2-4 are not found in standard IHC/ICC protocols and you might...we just made the sample bigger? In ExM, and its various derivatives, biological samples are enmeshed in...the number of specific targets you can label in a sample. Super-resolution microscopy (SRM), a family of... its original size. Depending on the protocol, samples can be expanded to ~4-10x linearly! This expansion...gel components to polymerize. Step 3: digest the sample to allow for even expansion.  In some cases, labels... ExM process allows you to physically magnify a sample so you don’t have to optically magnify quite so...

...green squares, purple circles and blue triangles. 2) An antibody immobilized on a bead binds specifically...protein G, a novel IgG-binding reagent. J Immunol 133(2):969-974. Hjelm H, Hjelm K, Sjöquist J (1972) Protein... adjusted depending on the sample. It is important to remember that sample lysates will also contain proteases..., incubate the sample as you would in an IP using a nonspecific antibody from the same host species as... from the sample prior to beginning the IP.  During the IP, the target protein in the sample binds to ...parallel with both a sample known to express the protein of interest, such as a sample transiently transfected...out in one of two ways. Both methods utilize the same basic strategy: immobilize the antibody or antibody-target...

...linear fashion. This implies that reducing the power 2-fold does not reduce photobleaching by half. How exactly...prolonged time). Other illumination strategies, such as 2-photon-excitation, selective plane illumination, TIRF...This can differ between fluorescent proteins in the same spectral class (Cranfill et al, 2016). This non-...comparison of several fluorescent proteins of the same spectral class on the microscopy system and in the...

...11 and Cas13 are 1). non-specific RNA cleavage and 2). cell toxicity. Cas7-11 does not display collateral...Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell, 163(3): 759-771 (2015). 10.1016...isolated which target and cleave RNA through the same mechanism as traditional spCas9. If you are looking...Cas13 and has been proposed to be useful for the same applications (RNA knockdown, RNA editing, etc.)....

...that the signal to noise ratio can be increased from 2-fold (by CRISPRainbow) to 3-fold with CRISPR-Sirius...CNV) analysis of the U2OS cells.   Figure 2: CRISPR-Sirius tricolor labeling. (A) Diagram of sgRNA-Sirius...combination of MS2, PP7, or boxB sequences in the same sgRNA will generate additional colors. Image from...

... you can read more about them in review articles (2–4). You can also take a look at my recent paper to...Development 118, 401–15 (1993). PubMed PMID: 8223268. 2. J. B. Duffy, GAL4 System in Drosophila : A Fly Geneticist...this mutant fly to one lacking the mutation in the same genetic background. In other cases, YFG or one of...

... Zetsche et al., Nat Biotechnol. 2015 Feb 2;33(2):139-42. doi: 10.1038/nbt.3149. Light-inducible ...and on your favourite signalling pathways (Figure 2). They are all available now at Addgene. 1 Lungu...20;19(4):507-17. 10.1016/j.chembiol.2012.02.006. 2 Guntas et al., Proc Natl Acad Sci USA 2015 Jan 6; ...Nihongaki et al., Chemistry and Biology. 2015 Feb 19; 22(2):169-74. High efficiency cloning for enzyme screening...inducible, gene knockdown with shRNA or shRNA-miR30, (2) tamoxifen-inducible CreERT2 gene deletion, (3) simultaneous...different fluorescent proteins, 8 constitutive promoters, 2 includible promoters, 3 polyA terminators and various...of plasmids required for each input/output system; 2) removing all chemically inducible promoters; 3) improving...

...scientists have 1) the courage to make mistakes and 2) the integrity to recognize these mistakes quickly...She studies beta-cell function and failure in Type 2 Diabetes. Birmingham, AL, USA 33.5207°N 86.8025°W ...available to people who want to join the dialog and 2) remember that science should not be partisan: everyone...campus speakers thoughtfully reminded us to use the same critical lens on our scientific community as we ... nor would you mention my small error and at the same time pass over in silence his very large one” (http...exoplanets into a successful PhD thesis. We use the same atmospheric physics to understand exoplanets that...happy to see so many others come out who feel the same.  I was also incredibly impressed with how well ...

...specific control of activity. Read More CRISPR Class 2 C lustered R egularly I nterspaced S hort P alindromic...plasmids. Plasmids within a given library have the same backbone, but they express or target different genes...

...expressed as a fusion with your favorite protein and (2) a synthetic fluorophore-contianing “ligand” that ...cell 58.4 (2015): 644-659. PubMed PMID: 26000849. 2. Grimm, Jonathan B., et al. "A general method to improve...removing excess dye from the sample, which is particularly important for samples where washing is difficult...reason is something called “the photon budget”: each sample has a limited number of fluorophores and each fluorophore...of information one can extract from a biological sample is wholly dependent on the photon budget and pushing... desperate to extract a few more photons from a sample. Chemical fluorophores can be substantially brighter...

...Table 2: Lab strains of E. coli Strain Natural resistance Primary Use Genotype ccdB Survival 2 T1R ...resistance to tetracycline Additionally, Table 2 provides a quick reference for some of the popular...convoluted due to researchers sharing and renaming samples throughout history. It was likely isolated in 1918...

...sites would confound your experimental results and 2) perform targeted off-target measurements at those...experiments where only localization of the Cas occurs. 2 gRNA approach Off-targets that come from mutations...perform downstream characterization of, isolating 2 or 3 distinct clones to perform key experiments on...If you are performing CRISPR library screens, the same principle holds true - the higher the number of ...

..., and the desired gene (transfer vector plasmid), 2) the HIV-1 genome containing all viral genes with ...protocols that have been described (protocol 1, protocol 2, protocol 3) to find the one that will give you the...PubMed PMID: 3785217. PubMed Central PMCID: PMC367857. 2. Naldini, Luigi, et al. "In vivo gene delivery and...are working with proto-oncogenes or with human samples that have not been tested for HIV.  If not using...ribosome entry site (IRES) and expressed under the same promoter. Alternatively, your gene and the marker...

...of ~75% in response to the magnetic field (Figure 2); they named this variant MagLOV. Andrew was thrilled...didn’t need any added cofactors.      Figure 2: Fluorescence intensity and ΔF/F of MagLOV, a variant...fluorescence when a small magnet was passed over the sample (Frank Hayward et al., 2024). The fluorescence ...intensity decreased whenever the magnet approached the sample (by less than 1%) and recovered after the magnet...expressing MagLOV while a magnet is waved under the sample. The imaged area measures approximately 82 mm across...Matsuoka, R., Kimura, S., Miura, T., Ikoma, T., & Kusamoto, T. (2023) Single-Molecule Magnetoluminescence...

... tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain. PLoS One. 2013 ...PMID: 24086725. PubMed Central PMCID: PMC3785459. 2. Krashes MJ, Koda S, Ye C, Rogan SC, Adams AC, Cusher...systems typically express Cas9 and a gRNA from the same plasmid. LentiCas9-blast virus can be used to generate...

...per cell increases 4-5 fold (for pUC plasmids) or ~2 fold (for p15a, R6K, and ColE2 plasmids) as cell division...biotechnology 36.4 (2018): 352. PubMed PMID: 29553576. 2. Kittleson, Joshua T., Sherine Cheung, and JChristopher...other hand, using a constitutive promoter in the same background resulted in a 20-fold variation of gene...

...connectivity to other data sources used in research; (2) connectivity to other tools used in research; and...by making it easy to get data out of the ELN; and (2) facilitate inter-operability with other research ...Github and Gitlab, and science-specific tools, e.g. sample management apps, colony management software, and..., not all of them support reproducibility in the same way. When choosing an ELN for your research, be ...commercialization community in greater Boston, while at the same time keeping one foot on the other side of the pond...

...estimate transformation efficiency (Figure 2).    Figure 2: Example of a serial dilution to estimate...library from Addgene, they’ll need to amplify the sample for further downstream applications. As individual... all of the bacteria can grow and compete in the same container, making it easier for a smaller, recombined...

...nonsense-mediated decay. Although Nelson et al. observed only 2% genome editing in one experiment, they found the ... cells throughout the body, especially stem cells 2. Remove any immunogenicity of the AAV vector Once ...Science. 2015 Dec 31. doi:10.1126/science.aae0169 2. Long C, Amoasii L, Mireault AA, McAnally JR, Li H...slightly different methods, they each observed the same positive phenotypes in cardiac and skeletal muscle...