...SCF-TIR1 ubiquitin ligase complex, promoting its interaction with the degron sequence and ultimately its degradation... plant protein TIR1 in non-plant cells, which interacts with the cells’ endogenous ubiquitin ligase, the...with mAID. Using this method, the lab was able to generate conditional alleles of a few essential proteins...attempted to engineer loops to create genomic interactions that wouldn’t normally occur. The synthetic ...enhancer. What makes LADL unique from previous iterations is the addition of CRY2, another Arabidoposis...

...encoded fluorescent executioner caspase reporters. Several apoptosis reporters have been developed, such as... and fluorescence based reporters however have several limitations. First, FRET signals are often difficult...They then optimized the length of both linkers to generate a reporter that would fluoresce: FlipCherry. FlipCherry...array of diseases and pathologies such as neurodegenerative diseases, autoimmune disorders, and cancer...Cabantous, Stéphanie, et al. "A new protein-protein interaction sensor based on tripartite split-GFP association...

...zebrafish, and mice. Second Generation iGluSnFR: SF-iGluSnFR In the second generation of iGluSnFR, the Looger... for Synaptic Imaging However, the first two generations of iGluSnFR had limitations: they were not optimal...extrasynaptic glutamate release. Building on the previous generation of the SF-Venus-iGluSnFR (itself an improvement...not neurons). iGluSnFR3 is also available in several ready-to-use viral vectors.    References and ...

...responsible for a phenotype can be identified by next-generation sequencing of the genomic DNA. However, some ...ApoB-NanoLuc reporter by inserting the NanoLuc luciferase downstream of the apoB gene. Using this reporter...approach to expand our understanding of ApoB-LPs and accelerate the discovery of additional genes and small-molecule...tool should serve as both a powerful tool for generating complex regulatory systems and further the study...nematode Caenorhabditis elegans. They have found several voltage biosensors and conditions that can be used...

...successful mutation of your favourite gene, there are several different methods available. Thankfully, there ...validation, such as this "CRISPR 101" post. Next-generation sequencing technologies are the gold standard...emerging validation technology. Digital PCR has several advantages over mismatch nuclease assays that will...desired genome edit. Since exploitation of NHEJ to generate translational reading frame-altering indels for...validation methods As mentioned, these assays have several major advantages over the commonly employed mismatch...

...cleavage promotes collateral cleavage of other RNAs. As with other Cas13s, this collateral cleavage does ...and Yan et al. describes new Cas13d enzymes that average only 2.8 kb in size and are easy to package in ...possibilities. To identify Cas13d, both groups used very general bioinformatic screens that looked for a CRISPR ...accessory protein RspWYL1 increases both targeted and collateral RNA degradation by RspCas13d. RspWYL1 also increased...

...to edit. These cells are then transfected with several different constructs for the reporter to work and...your CRISPR experiments. But how? There have been several systems to do this but most are labor-intensive... the native methionine start site. This first iteration was named GEmCherry1, due to the “genomic fragment...and 21 to create GEmCherry2. The Denham lab made several other small changes creating 2 other versions but...  Find GEmCherry2 reporter plasmids at Addgene Overall SRIRACHA and GEmCherry2 reporter systems provide...

...the target molecule. Although it’s possible to generate antibodies against many types of molecules including...recognize proteins. Figure 1: Steps in the generation of hybridomas for monoclonal antibody production...19420862.2018.1445456  Holzlöhner P, Hanack K (2017) Generation of Murine Monoclonal Antibodies by Hybridoma ...(DC): National Academies Press (US); 1999. 1, Generation of Hybridomas: Permanent Cell Lines Secreting...hybridomas  Learn about monoclonal antibodies for immunotherapy Resource on Addgene.org Check out Addgene’...

... a structural precursor protein, and Pol is a polymerase. Envelope gene (may be pseudotyped to alter infectivity...γ-retroviral packaging plasmids are not interchangeable. General envelope plasmids, such as VSV-G (Vesicular Stomatitis...packaging cell line. For example, Phoenix, a second generation γ-retrovirus packaging cell line developed by...region of the 3’ LTR and are primarily used in gene therapy research to reduce promoter interference and/or...More resources on addgene.org Addgene Protocols: General Transfection The Addgene Retrovirus Guide  ...

...single color controls On a flow cytometer, you generally want your positive cells to read between 104 and...provided by the antibody companies or check the literature. Pro tip! Early gates, like your live/dead markers...population (b). There is some debate in the literature as to when you need isotype controls. Our lab...isotype controls, while others don’t. However, the general consensus seems to be that if you are looking for...research background in CD8+ T cells and cancer immunotherapies. She is currently a research technician in ...