... Genes Dev. 1988 Jun;2(6):718-29. doi: 10.1101/gad.2.6.718. PMID: 2843425.Takara Bio. Tet-On and Tet-Off...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... the system have been engineered and improved. Takara’s “2nd generation Tet-Off vectors (Tet-Advanced)...this way, you can interrogate the activity of uncharacterized promoters. Similarly, placing UAS upstream ...-Off systems: second generation. https://www.takarabio.com/products/gene-function/tet-inducible-expression-systems...

... pCRISPomyces-2 contains Cas9 and a gRNA cassette. The simpler system of pCRISPomyces-2 displays a higher...and a subsequent curing protocol that requires only 2-3 hours incubation. E. coli and T. citrea scarless...four major components including (1) a CRISPR RNA, (2) four proteins forming the QCascade DNA-targeting ...expresses lambda Red and Cas9, which are separately inducible by L-arabinose and anhydrotetracycline (aTet), respectively... template or non-template strand, among other parameters. .  Stanley Qi lab's CRISPRi plasmids Stanley...across multiple gRNAs per gene (Singh et al. 2016). Sarah Fortune's lab M. tuberculosis CRISPRi plasmids Noting...proteotoxic when used in Mycobacterial species, Sarah Fortune’s lab screened 11 other Cas9 orthologs to...

...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...highest-ranking predicted off-target sites. Each paper characterizes additional, unique facets of CRISPR DMD therapy...effectiveness and off-target risk must be evaluated separately, increasing the time to clinical approval. Even...Blog Learn How CRISPR Could Be Used to Treat Cataracts and Cystic Fibrosis Read How Homology Directed...

...wisdom  Reach out to 2-3 individuals in your professional circle to vouch for the character you painted in ...and scholarships in different subject areas!    2. Resume and cover letter know-how  Tailor your resume...increase your competitive edge, they alone, are no guarantee of success. Don’t luck into an internship, prepare...

...episode 2 "Organizing Literature" : I found out about Zotero here Dear Grad Student podcast episode 2 "Essential...flagging passive voice, overuse of adverbs, and paragraphs with many hard-to-read sentences.  Grammarly ...

...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...al, 2017) and cannot be described by a single parameter. Therefore, it is necessary to know how the fluorescence... power, the photostability of the variants is comparable (lower panel of figure 1). To avoid changes in...microscopy (University of Amsterdam). He develops, characterizes and uses genetically encoded fluorescent probes...

...studies have shown that the conditions Sotos-type 2 and Marshall Smith Syndrome are due to mutations in...accurate diagnosis of NFIX-related syndromes (Sotos-type 2 and Marshall-Smith) and could be expanded to NFIA-...manifestations. PMM2-CDG is the most frequent CDG type characterized by deficient phosphomannomutase (PMM) activity...suffer from immunological defects. These are characterized by a predisposition to severe and recurrent ... in the NFIX gene. These diseases are characterized by overgrowth phenotypes and neural developmental ...

...   1)   “Piled Higher and Deeper” by Jorge Cham   2)   “Boxplot” by Maki Naro   3)   “Bird and Moon” by...Unearthed Comics” by Sara Zimmerman   5)   “Beatrice the Biologist” by Katie McKissick       Sara Zimmerman, Unearthed...

...reach out to the corresponding potential depositors. 2. Tweet at us! Tweet @Addgene with #suggestaplasmid...where we can find the plasmid. While we cannot guarantee that everyone we reach out to will end up depositing...

..., 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...safer system. Unlike the MoMLV, lentiviruses, a separate genus of the Retroviridae family including human...facilitates nuclear export) is expressed from a separate plasmid, and the promoter of the 5’LTR has been... In this case your gene and the marker can be separated by an internal ribosome entry site (IRES) and ...

...residues, which can be found throughout the antibody. 2) Endogenous cysteines, such as those that make up ...Detecting antigens by quantitative immuno-PCR. Nat Protoc 2:1918–1930. https://doi.org/10.1038/nprot.2007.267 ...commercially conjugated antibodies available, that never guarantees the antibody you need will be conjugated, or ...//doi.org/10.3390/antib4030197  Hegazy M, Cohen-Barak E, Koetsier JL, et al (2020) Proximity Ligation ...

...ITR, which can either be “flip” or “flop” (Figure 2). Additionally, the ITR contains a 4-nucleotide Rep...site for Rep proteins (Savvy et al. 2013). Figure 2: AAV2 wild type ITR in flip and flop orientation. ..., B-B’, and C-C’) that give the feature its characteristic T-shape. The arrangement of the B-B’ and C-...

...misplacing your results or losing that lab notebook from 2 years ago. The free subscription to Lab Archives allots...notebooks you’re saving trees! The Earth thanks you. 2. Gibco Cell Culture by Gibco Whenever I start to learn...MSDS for a lab chemical of choice. Now, I can’t guarantee every reagent in your chemical closet is in this...

... reduce the background staining (Table 2).      Table 2: Secondary antibody selections for a four-antibody...with users trying to probe multiple targets in parallel. In these cases, it is critical that users choose... experiment.  When probing multiple targets in parallel, use the most narrowly specific secondary antibodies... four-antibody panel, staining in parallel    In short, when staining multiple targets, look for primary...

...days before the deadline isn’t going to be much use. 2. Originality is essential for grant success The two... it’s important not to throw in the towel if 1 or 2 funding bodies say no. If possible, get feedback from...perfectly reasonable paragraph unintelligible to a lay audience. My lay paragraph typically includes a...kindly friends and family to read my non-expert paragraph and point out words or sentences that are unclear...

... signal back again. The Ting lab constructed 2 versions of this reporter to target the K9 (pcDNA3-K9 ... 525.7570 (2015): 533-537. PubMed PMID: 26352475. 2. Bygren, Lars O., et al. "Change in paternal grandmothers...disrupts normal function. Angelman syndrome is characterized by intellectual and developmental disability...or smiling, whereas Prader–Willi syndrome is characterized by low muscle tone, short stature, cognitive... in the FRET signal. Subsequent demethylation separates the fluorescent units and so lowers the FRET signal...

...populations that may otherwise be difficult to separate out. In Figure 2, you can see that, without the FMO, it... from the Ly6C positive population. Figure 2: An FMO gating example. The FMO sample (A) can be ...

...represent a favorable state for tumor progression (Ref 2). Future treatments may need to target this immunosuppressive...cell 30.1 (2016): 147-160. PubMed PMID: 27411589. 2. Todoric, Jelena, Laura Antonucci, and Michael Karin...the Relationship” by our guest blogger Subhadra Jayaraman, a doctoral candidate at Binghamton University...

... 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...

...knock-out cell line to perform downstream characterization of, isolating 2 or 3 distinct clones to perform key...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...