... dementia-associated tau, delivering dCasRx and a 3-spacer array targeting the MAPT exon 10 splice acceptor...for pre-clinical and clinical applications of dCasRx. The identification of Type VI Cas13d enzymes is ...RspCas13d. RspWYL1 also increased EsCas13d activity, indicating that WYL domain-containing proteins may be broader...even when guides were tiled across a target RNA, indicating that this enzyme does not cleave at a predictable...target essentially any RNA in a cell. Since catalytically dead dCasRx maintains its RNA-binding properties...

...spCas9 from Streptococcus pyogenes, this sequence is 3’-NGG. But Cas9s found in other species can recognize...to generate a genomic alteration at a specific location, it must be accessible by Cas9. Our previous blog...off-target activity, which may be undesirable in many applications. At the time of our first blog post on choosing...CRISPR-Cas9 to increase its specificity." Nature communications 9.1 (2018): 3048. PubMed PMID: 30082838. PubMed...CRISPR-Cas12b for human genome editing." Nature communications 10.1 (2019): 212. PubMed PMID: 30670702. PubMed...

...transfection. The other (mKate2) is regulated in the 3’ UTR by four miRNA target sites and thereby serves...change depending on the location. Current methods for visualizing the location and function of microbes...that undergo light-induced dimerization. Upon application of blue light, iLID undergoes a conformational...discovered in 2014 that these gas vesicle proteins scatter sound waves and create ultrasound contrast. At ...

...sequences more efficient and less risky.   Fig. 3: Development of a cleavage-free dCas9-based gene editor...of interest and, for in vivo work, generate a dedicated transgenic line. Using zebrafish as a model organism...off-target mutations. These plasmids contain catalytically inactive dCas9 and microbial SSAPs (dCas9-SSAP...clonal multiplexing of cells in live imaging application. Each visual barcode consists of a fluorescent...transcription response element (TRE) or kinase translocation reporters (KTRs). Find visual barcode plasmids...

...PMID: 21364278. PubMed Central PMCID: PMC3069789. 3. Kügler S, Kilic E, Bähr M. Human synapsin 1 gene ... preps. Cas9 is the headliner of most CRISPR applications and can be expressed from your cell line of ...Find Ready-to-Use Lentivirus Find Ready-to-Use AAV Catch up on Chemogenetics ...

...ssRNA engaging one strand of a dsDNA molecule; the 3-stranded structure produced is known as an R-loop....generate dsRNA as part of their infection and/or replication process within the host. dsRNA viruses are inherently...generate dsRNA intermediates as part of the viral replication process. dsDNA viruses yield dsRNA through a ...

....  For example, the phosphorylation status of SRC-3 defines its association with other members of the ...murine members. NRs permit the integration and communication of such signals between central and peripheral...macrophage activation. Our mass spectrometry-affinity purification proteomic profiling of coregulators in HeLa ...

...and complete screening in just 3 days? Thanks to Tyler Jacks’ lab, this 3-day goal is is now attainable...construction of a so-called “genomically recoded organism” (3) but its portability to other bacterial strains remains... the assembly of TALE repeats into a single-tube, 3 day process by utilizing pre-assembled trimers to ...FAIRE-seq, and CHIP-seq), genome-wide identification and quantification of enhancer activity have remained...compartments (cytosol, Addgene catalog #71245; mitochondria, Addgene catalog #71247) Rosella provides an ...CpG methylation by fusing catalytically inactive Cas9 (dCas9) with the catalytic domain from DNA methyltransferase...  Superglue proteins with SpyTag/SpyCatcher and SnoopTag/SnoopCatcher The development of recombinant DNA...

...and best characterized of these is the Src homology 3 (SH3) domain family, known to be involved in cytoskeletal...interactions of proteins within these structures. Enzyme-catalyzed proximity labeling (PL) uses a promiscuous labeling...

...10) and grow them in a small culture volume (2 or 3 ml) containing the corresponding antibiotic, extract...constructs. His work was acknowledged in hundreds of publications including Cell, Nature, and other journals. ...

...ankyrin repeat proteins. DARPins typically consist of 3-4 repeats, have high specificity and binding affinity...re used in a number of research and clinical applications. Most of the time, when someone speaks about...deeper penetration into a tissue, useful for applications like IHC, or be able to access an epitope folded...that would size-exclude an antibody. In imaging applications, smaller affinity reagents means higher resolution...antibody’s ‘Y’. They are often used in clinical applications. Fab fragments can be created in lab in situations...Shahied, 2004). They are often used for clinical applications, including imaging.  Diabodies are ~60 kDa proteins...antibodies are still an excellent option for many applications, and frankly, if ain't broke, don't fix it!  ...

...enter the cells so they can go to work. Figure 3: Inducing the double strand break and introducing ...visit the Cell Catalog on the Allen Cell Explorer, where you can also find the certificate of analysis provided...signal that makes it hard to visualize the precise location of the protein or structure of interest. Endogenous...technology, inserting large tags into a precise location in the genome is still inefficient, particularly...from these cell lines can be found in the Cell Catalog on our data portal. The plasmids should be functional...

... to have either salaries raised or hours tracked, 3% of postdocs were at institutions focused only on ...days prior to the implementation date, our data indicated that, while 69% of all postdocs were expected ...sent, with the lack of transparency and poor communication of their plans (many postdocs - and in some ...to reach out to institutions with a statement indicating that we encourage them to keep raising salaries... little correlation with cost of living at the location of the institution. Continuing efforts to gather...Director of Future of Research. He is interested in advocating for and with junior scientists, to help them ... Bring Increased Postdoc Wages Read about the Advocating for Science Symposium and Workshop Learn about...

...chains. 3) Endogenous carbohydrates added to antibodies post-translationally. 4) Modifications made to...These properties allow for efficient signal amplification, making biotin conjugated antibodies good options...especially if you start to consider clinical applications). But there are a few other classes that we ...proximity ligation, immuno-PCR, and single-cell applications (Hegazy, et al., 2020; Niemeyer, et al., 2007...also conjugate antibodies directly to beads for purification purposes.  Coupling up - antibody conjugation...Detecting Protein-Protein Interactions and Protein Modifications in Cells and Tissues in Situ. Curr Protoc Cell...

...PMID: 24529477. PubMed Central PMCID: PMC4139937. 3. Anders, Carolin, et al. "Structural basis of PAM-...As evidenced by all the CRISPR publications, press, and plasmids out there, it’s obvious that CRISPR ...nickase variant, lowering Cas9 expression, and truncating gRNA sequences used for targeting; however, these...on-target and off-target activity at multiple genomic locations in HEK cells. Further testing of two of the mutants...PubMed PMID: 26628643. Find plasmids from this publication at Addgene. 2. Nishimasu, Hiroshi, et al. "Crystal...PubMed PMID: 26735016. Find plasmids from this publication at Addgene. 5. Chen, JS, et al. "Enhanced proofreading...PubMed PMID: 28931002. Find plasmids from this publication at Addgene. Resources on the Addgene Blog Read...

...PMID: 26258682. Find plasmids from this publication at Addgene. 3. Kirchhofer, A., et al. “Modulation of...repurpose popular GFP reporter lines for more sophisticated experimental manipulations, saving the time ..., 98% of TdTomato expressing cells were GFP+, indicating a robust, but specific system. T-DDOGs also successfully...systems are easily adaptable to neuroscience applications, including optogenetics, and they should make...Could Cas9-DOG be developed soon to allow genome modification only in a small subset of cells? Other proteins... PMCID: PMC4096992. Find plasmids from this publication at Addgene. 2. Tang, Jonathan C.Y., et al. “...

...coaching. Always ask for references and follow up with 3-4 to hear about those clients’ experiences. Good coaches...choose well. What are their qualifications? Do they have certifications? How can they demonstrate their...  Check out Joanne's Reddit AMA I am a strong advocate for peer mentoring, but volunteer mentors are ...help you. A good coach usually will be a clear communicator and make direct suggestions for improvement ...coaching is nice if you can get it.  Because I am located in Cambridge, MA (and because we have a lot of ...she is especially excellent in coaching for communication skills. Industry transition is also an area ...

...one locus (e.g. Cre) and heterozygous at the other. 3. Breed your F1 generation Now, your goal is to generate...techniques that can be used to introduce genetic modifications into mouse embryos. But once you generate a ...techniques are each useful for introducing one modification into the mouse genome, they are not commonly...parental mice In order to ensure that your genetic modification will be passed along to all of the offspring...environment and will harbor similar epigenetic modifications. Special breeding cases Whether your mouse colony...the mutations that you are trying to cross are located on homologous chromosomes in the parental generation...

...endonuclease of a class 2 CRISPR-Cas system. Cell, 163(3): 759-771 (2015). 10.1016/j.cell.2015.09.038 Harrington...a defined site to form a clean DNA break, Cas3 locates its target sequence and initiates single stranded... out large regions of DNA and has anti-viral application potential as a result. Fast facts – PAM requirement... has been proposed to be useful for the same applications (RNA knockdown, RNA editing, etc.). The main...

...Genetic Analyses in Mice. 2004. PubMed PMID: 14723844. 3. Atasoy D, Aponte Y, Su HH, and Sternson SM. 2008....sites directs the Cre recombinase to invert, translocate, or excise a DNA fragment. The availability of... the function of various point mutations and truncations in the absence of the endogenous gene. Spatio-temporal..., allowing for clearer characterization of the location, morphology, and circuit mapping of a targeted...