...based gene editing. But, how do you decide which tools to use, and what do you need to know about them ...similar to what the endogenous binding partner would do. This allows precise control over when and where ...

... human genome (Munoz-Lopez et al., 2010).  So why do transposons matter? The outcome really depends on...how retroviruses, like HIV, replicate. Class I TEs do not encode a transposase enzyme.  Class I TEs are...TEs are also known as DNA transposons, since they do not use an RNA intermediary when they move. Most ...

...non-coding RNA, cutting most anywhere within the RNA will do. Similarly, if you want to knock out a gene, a frameshift...introduced anywhere within the early coding region will do the job. What matters for these experiments is cutting...

...tool for gene delivery, but it has a drawback: how do you ensure your gene goes where you want it to? Knowing...approach to enhance specificity in retinal cells. To do this, the group developed a new strategy for finding...

... . https://doi.org/10.1038/srep31730 Tsuruta LR, dos ML, Moro AM (2018) Display Technologies for the Selection... 2015a). With molecularly defined antibodies that do not change over time, scientists will know exactly...Harbor Laboratory Publications, New York. Helma J, Cardoso MC, Muyldermans S, Leonhardt H (2015) Nanobodies...

...sgRNA. What if I want to work with gene drives? Then do so! It's a tremendously promising field that could...degree of confinement are those of the author and do not represent a formal stance taken by Addgene or...

...everyone gets involved. So join the conversation! Do you have a question about culturing iPS cells? Confused...

...detection range of your protein. The most common way to do this is via a Bradford assay or BCA assay. Be sure...preserve specific protein modifications, we recommend doing a little more research into gel chemistry and/or...wide dynamic range for fluorescent proteins. They do tend to have increased background with higher exposure...your blot to calculate the amount in your sample. (Do ensure your blot is optimized so your standard curve...

...our favorite posts from 2022! Wow! Our depositors do some really incredible science - I hope you've enjoyed...

...biochemistry researcher at New England Biolabs. When do you need a high fidelity ligase—and when is an alternative...base pairing through minor grove contacts, and thus do not read specific base sequences, but are sensitive...differential is not known for certain, but may have to do with the slight melting of the 5 ́-terminus during...

... a part of a community that recognizes doing science (and doing it ethically) runs hand in hand with policy...views represented below are those of the writers and do not necessarily represent the views of Addgene. ...

...fluorescence when excited with ultraviolet (UV) light, they do so via aromatic amino acids found in all proteins...photons excite electrons of a fluorophore, they can do so at a range of wavelengths, rather than at just...1, emission spectra of neighboring colors can and do overlap. For example, APC overlaps noticeably with...

...they are present at a very low concentration. So how do we make sure to catch all of these potential mishaps...expected hits, we always get hits to “random” genes. Do these hits always mean the sample is contaminated...users titrate their AAV vectors to find the optimal dose that allows for sufficient transgene expression ...

...use. This will prevent spills, but, if any spills do occur, they should be wiped with 70% ethanol immediately...morphologies, being familiar with these morphologies and doing a quick check under the light microscope will help...

... = 500,000 colonies.  Pro tip! Since you probably do not want to individually count 500,000 colonies, ...did not result in significant recombination. If you do see recombination, we recommend ensuring the cells...

...use But exactly what kind of editing are people doing with all these CRISPR plasmids (and libraries)? ...scientists use CRISPR technology? We think so — and so do our requestors! CRISPR plasmids in the Addgene repository...

...newer, such as the relatively recent resurgence of “Do-It-Yourself Science” described in Chapter 3, and ...never inherently bad or evil. However, it is easy to do sloppy science, misrepresent science, or misuse science...

...Addgenies about their career path. Graduate school: Do I or don’t I? I asked the nine Addgenies about why...told her that if she found something she loves to do, she would never work a day in her life. She said...

...coronaviruses and enable the virus to enter host cells. To do this, they injected the spike protein from two different...effectively to protein targets, similar to how antibodies do. The structure of the common antibody is heftier ...

...guanines). This limits the use of ABEs as DNA sites that do not contain a suitable PAM cannot be edited efficiently...While base editors are precise molecular tools, they do have undesired off-target editing effects. ABEs have...