...users have recently begun to develop CRISPR-based approaches for studying gene function in these systems. ...use in a fetal-lung derived organoid system, the approach is intended to be amenable to organoids derived...tissue-types. By bringing together CRISPR-based approaches and organoid models, toolkits like this one provide...trigger seed germination of parasitic plants, this approach was also used to build and validate a ratiometric...parasitization of cereal crops. As a result, agricultural applications of these novel biosensors could have a major...

...isolating the cell line all the way to modern applications. What the HEK is it?   HEK cells were first...cell biology, there are thankfully many other applications that they are useful for! What the HEK is it...cell line and we will highlight some of those applications here.   Virus production HEKs are the go-to ...proteins at high levels. In a case where clinical applications of recombinant proteins are necessary, HEKs ...significantly reduces the cost of production. Research applications There are two areas of research where HEK cells...

...NgAgo replace CRISPR? Such a drastic shift won’t happen overnight, but there are a few reasons why you ...flexibility in choosing a genomic target. NgAgo also appears to outperform Cas9 in GC-rich regions. DNA guides...staggered (Cpf1) cleavage. NgAgo takes a different approach, randomly removing 1-20 nucleotides from the cleavage...site specified by the gDNA. Depending on your application, this could be favorable or unfavorable. In a...activity. One-guide faithful NgAgo guide loading appears limited to the period when the protein is being...

...generation of various flux imbalances. The conventional approach has typically been to either systematically delete...networks and multiplex gene control. However, these approaches are often met with the challenges of regulatory... possible. From a design perspective, this is appealing since the RNA scaffold modules are comparatively... multiplex control via scRNA design is readily apparent in metabolic engineering. The authors elegantly...control of gene expression. This strategy can be applied to effectively change metabolic flux. The dCas9...

...axial dimension that will appear in focus). This means that the images will appear sharp without interference...excite a molecule of GFP. Because red light has approximately half the energy of blue light, two photons of...density of red-photons be establish for this to happen.Using red light has a second advantage: red light ...objects lying within the diffraction limit will appear as one object in the final image. Over the past...of light – think of a single GFP molecule – will appear as a fuzzy 200-300 nm spot when imaged. Super-resolution...

...IVT sgRNA) approach (Dewari et al. 2018 and Figure 2). Not only is the cr:tracrRNA approach highly efficient...biochemical tasks in the cell. One of the most popular approaches to study protein interactions is the pull-down...disrupts protein function, an N-terminus tagging approach should be pursued, however, clonal lines must ...me at [email protected]. Keep pipetting and happy genome editing! Pooran Dewari is a postdoc in Steve...optimizing CRISPR/Cas9 genome-editing methods and applying them to study brain tumour biology. When not pipetting...

...neurons) that can be turned on or off by the application of a small molecule ligand. The ideal chemogenetic...translational studies because it is FDA and EMA approved. One pitfall of a suite of muscarinic-based DREADDs... develop a PSAM that responded to a clinically approved drug, the PSAM domain was engineered to respond...Azua IR, Li JH, et al. (2009). A chemical-genetic approach to study G protein regulation of β cell function...chemogenetics for research and potential clinical applications. Science. 364(6436):eaav5282. PMID: 30872534...

...the light source. Optogenetics has been broadly applied to study the physiology of the brain and nervous... peak expression of the opsin is reached. This approach is especially powerful when combined with site...conformation/dimerization. In addition to the applications described above, both LOV and Dronpa can control...KR, Deisseroth K. 2010 Molecular and cellular approaches for diversifying and extending optogenetics. ..., Yizhar O, Deisseroth K. 2011. Principles for applying optogenetic tools derived from direct comparative...

...have high levels of off-target effects, both of happen with Cas13. CRISPR-Csm degrades both nuclear and...antibodies may just not perform as well in one application as another, so it is always best to have options...for mEGFP-HRas (from Plasmid 18662). See the Applications section on the Anti-GFP [N86/8R] page for more...fluorescent proteins (FPs) necessary for many of these applications have lagged. Fixation, chemical staining, and...well-suited for the demands of advanced microscopy applications such as super resolution, electron, and expansion...

... of applications with the potential to save lives. While most scientists recognize and appreciate the ... Yamanaka himself, during his keynote address, appeared almost incredulous that the field has exploded...daunting because they have no clear cut answers, I applaud the stem cell community for addressing these important...but they are also the reason this research is happening. Stem cells scientists have certainly not lost...

...interactions. Beyond these common applications, researchers are finding new ways to apply fluorescent proteins everyday...Zhou and Fatima Enam. There were many excellent applicants but Xin and Fatima stood out for their creativity...Light-inducible Proteins (FLiPs) Fluorescent proteins can be applied in a long list of creative ways. In her graduate...utility of these FLiPs in a variety of research applications and particularly notes their ability to be precisely...

...altering its function). HDR is crucial for certain applications, for example, correcting the allele that causes...this question led us to develop a new, rational approach to improve the efficiency of sequence replacement...introducing a cut, with residence times on DNA of approximately 6 hours. To our surprise, we measured identical...region allowed sequence replacement rates of approximately 1%. This is by no means a large number, but ... order of magnitude greater than transfection approaches. Moreover, the cost of reagents is relatively...

...together a grant application? Are you familiar with the publishing or patent application processes? How ...How can you stand out among all the candidates applying for the position?   Here are a few things to consider...makes sense for your skills, and the job you are applying for. Also remember that a resume is typically ...have done is closely related to the job you are applying for, in which case talk about your research, as...

...disciplines. In biology, innovative new imaging approaches are constantly being developed which combine ...biological protocols and revolutionary physical approaches. However, with each new technical achievement... greatly increases accessibility. Through this approach, the Planktoscope, a portable device for studying...on-going creativity with the construction plans. Appropriate open-source licenses for hardware (e.g. CERN ...optical setups. References Joel T. Collins, Joe Knapper, Julian Stirling, et. al, "Robotic microscopy for...

...advances and how some of these variants may be appropriate for your specific research question. Considerations...off-target activity, which may be undesirable in many applications. At the time of our first blog post on choosing...Using a combined random and directed mutagenesis approach, the Anna Cereseto Lab created pX-evoCas9, which...enzymes may better suit specific experimental approaches. One example is CasX, characterized by the Oakes...While no enzyme is perfect for all experimental approaches, knowledge of both the advantages and limitations...

... by the Dueber Lab at Berkeley, provides a new approach for generating novel genetic variants in bacteria...the key components of EvolvR and its potential applications. Challenges of existing mutagenesis methods...exist, but there are a few drawbacks with these approaches that makes it challenging for users to achieve...is capable of mutagenesis in E. Coli, but this approach requires special bacterial growth conditions in...more than one mutation near the target site. Applications of EvolvR The Dueber Lab used EvolvR to discover...

...multicolored nano-lanterns for use in various applications, including optogenetics, biosensors, and fusion...furimazine. In addition to the previously described applications, the enhanced signal of eNL allows their use...characterizing Nano-lanterns and eNL for many different applications. For long-term imaging, especially where photobleaching...concern, these constructs come in very handy. This approach is especially suited to complement optogenetic...Therefore, especially thick samples may not be appropriate for use with Nano-lantern technology. Multicolor...

...principles we used in our daily content writing applied to alt text. We also were aware of the need for...they hoping to get from it? The image in Figure 1 appears on our protocol page for AAV Purification by Iodixanol...a lot of jargon and have a technical, detailed approach. Other times, it is more helpful to use plain ...start small with alt text or immediately dive into applying these four steps elsewhere, engaging in this practice...from posters to papers.  We hope this helps, and happy writing!   Resources Additional resources on the...

...next-gen sequencing. ChIP is an incredibly useful application for exploring these interactions, and can give...including epigenetic modifications. But it's not an approach for the faint of heart!  The Process ChIP can...preferred method, enzymatic digestion can be appropriate when your protein of interest is a low abundance... upfront investment than many other antibody applications, this investment can well be worth it once you... idea to add "patience and an extra-methodical approach" to the top of your protocols when embarking on...

...some reporters are better suited for particular applications. Read on to learn more about each reporter or...sensitive than HRP RANbody detection, it is approximately as sensitive as detecting the same antigen with...less sensitive than HRP RANbody detection but approximately as sensitive as antigen detection with a standard...detected with a labeled anti-HA or -MYC antibody, approximately as sensitive as detecting antigen with a standard...Detection with an anti-Chicken IgY antibody is approximately as sensitive as detecting antigen with standard...