...will be suitable). For N-terminus tagging, the promoter region of both the alleles should be sequence-...cautiously analyzed for any damage (indels) to the promoter region of the non-HDR allele. For N-terminal tagging...cuts 8-15 bp downstream of Stop codon; for N-terminus tagging, a crRNA cutting 8-15 bp upstream of your gene...Dewari, a postdoc in Steve Pollard’s lab at the MRC Centre for Regenerative Medicine (CRM), Edinburgh. Most...universal all-purpose “tag antibody” can be used for downstream IP/ChIP assays. Creating epitope-tagged clonal... is a postdoc in Steve Pollard’s lab at the MRC Centre for Regenerative Medicine (CRM), Edinburgh. His...

...microscope and GFP fusion proteins controlled by promoters specific to the immune lineage, scientists could...Finsterbusch, Michaela, et al. "Patrolling monocytes promote intravascular neutrophil activation and glomerular...that T cells were able to extravasate from the bloodstream to invade lymph nodes and scan antigen presenting...

...characterize the function of certain regions in a promoter or gene, as well as to study the effects of inactivating...methods, chances are you’d like to find a better one. Trehan et al. wondered if, instead of relying on in vitro... each other; ~17 bases is optimal. After being treated with DpnI to digest away the template plasmid, ...applies to deletions; using REPLACR-mutagenesis, Trehan et al. deleted a whopping 144 kb from a bacterial...recombination. Across all modifications described in Trehan et al., REPLACR-mutagenesis’ median efficiency ... for site directed mutagenesis   References 1. Trehan, Ashutosh, et al. (2016). “REPLACR-mutagenesis:...

...fluorescent proteins and is driven by an inducible promoter (Plasmid 64542 or 64543) Before translation, ...cellular function, especially during development or stress. With ribosome profiling, researchers have been...

...forms as part of graduation, hiring, funding, and promotion processes, I’m confident that everyone’s enthusiasm... review and primary research journals including Trends in Cell Biology, Cell, Molecular Cell, Developmental...

... and funders in the Transparency and Openness Promotion Guidelines; preregistration to increase clarity... order, or every combination of materials and treatments is all too often glossed over. This is why it...which data are investigated to look for unexpected trends or differences between groups. Hypothesis generation...hypothesize that growth rates will be higher in treatment versus control, and you think there are four important...

...and tech transfer communities who help guide and promote Addgene in its mission to make research easier.... interesting biological questions, you need a tremendous amount of technology development alongside it...what is worth having. The work that [Addgene’s] outreach scientists do to comb through the literature, ...

...the Francis Crick Institute concluded “Aspirin promotes autophagy, inhibits platelet aggregation, and ... that fasting has a positive effect on cancer treatment (Ref 1). As described in the presentation, starvation...favorable state for tumor progression (Ref 2). Future treatments may need to target this immunosuppressive reaction...

...visualization and expressed from a heart-specific promoter. Fish were labeled with MG-2I, either by incubating...hearts 24 hours post treatment and no mCer3 cells present 96 hours post treatment (see figure 2). Induced...culture experiments, only dL5**-expressing cells treated with MG-2I dye and exposed to light stained dead...Chromophore-assisted laser inactivation in cell biology. Trends in Cell Biology,18(9), 443-450. doi:10.1016/j.tcb...

...assembly reaction. DNA fragments with the desired promoter, selectable marker, and target gene, for example...and improvements in these methodologies offer streamlined and simplified alternatives to traditional cloning...

...studying how that one protein interacts with that one promoter can tell you, it can be easy to get lost in the...Fight Rare disease with iPSCs Read About Addgene's Outreach Efforts in the Microbiology Community Sign Up ...

...Expression is driven by a fairly strong EF1alpha promoter. We have verified expression of correctly-sized...straight-forward. All constructs are tagged with a 2xStrep-tag for easy detection and affinity purification...

...changes were introduced to nCas9. These changes promote nCas9’s dissociation from DNA after nicking and...bacteria resistant to both spectinomycin and streptomycin, but could also be used for studying epistatic...

...recognition of transcription-associated DNA damage that promotes homologous recombination. Gong F, Chiu LY, Cox...nucleosomes, loops, chromosomal domains, or how stretches of DNA on different chromosomes interact with ...K, Wang W, Rando OJ, Xue Y, Swiderek K, Kuo A, Crabtree GR Cell. 1998 Nov 25. 95(5):625-36. An architectural...

...Viral expression cassettes typically contain a promoter (inducible or constitutive), your gene of interest...the last decade, viral applications in cancer treatment and gene therapy have also been in the limelight...approved the first oncolytic viral therapy in 2015 to treat melanoma, and many more are in the pipeline. Viruses...

... 2008). Obligate dimers (such as dTomato) that promote interactions between the FPs interfere with native...Section of Molecular Cytology and Van Leeuwenhoek Centre for Advanced microscopy, University of Amsterdam...applications (e.g. labeling of cells or tracking promotor activity), but it is problematic in applications...Section of Molecular Cytology and Van Leeuwenhoek Centre for Advanced microscopy (University of Amsterdam...

...-loops described above play distinct roles in promoting cellular processes and functions, but they also...production. In the same vein, genotoxic stress, such as chemotherapy treatment or ionizing radiation, can also...

...complex, to temporarily inhibit this process and promote copying of the edited strand to the unedited strand... co-express mismatch repair inhibitor MLH1dn Promote desired resolution of heteroduplex     ✓ ...non-homologous end joining. As a result, Cas9 treatment causes most products to be indels while the efficiency... In addition to the numerous innovations that streamline the process of prime editing, the Liu lab contributed...efficient as possible. Figure 2: Decision tree for selecting a PE6 variant. PE = prime editor; ... 2023) Prime editing has a lot of promise for treating genetic diseases. In our summer Hot Plasmids post...

...several weeks which may also depend on transgene or promoter choice. Behaviors you can study using viral vectors...into the experimental setup, the mouse becomes stressed. It will likely move around faster in the following...some time to adapt to a testing apparatus. General stress levels of the test animal may also be reduced by...noise should be avoided as this will also increase stress levels and affect behavior. A fact to consider:... groups), health status, diet, cage enrichment, stress level, origin (vendor or in-house mouse breeding...

...nuclear localization, binding to the appropriate promoter sequence and activation of gene expression. In...bulk cell experiments, Clift et al. observed no stress or damage upon TRIM-Away, and the electroporated...re-adhered to the culture plate within 4 hr of treatment. In some primary cell lines, endogenous TRIM21...Listen to the Fire-Cas9 Podcast Segment Jerry Crabtree’s lab has developed a new CRISPR/Cas9-based system...