...donor-acceptor pair and a change in FRET efficiency correlates with a change in the distance and/or orientation...available. Fortunately, several excellent reviews have collated data from various papers to make the decision ...conformation of the probe. More on these biosensors later on). Experiments using intermolecular FRET probes...simultaneously would further increase the amount of correlated information available from a cellular process...

...embark on or continue their careers in science-related fields. In particular, two recent events have the...note is a ruling in August by the National Labor Relations Board to allow graduate students who work as teaching...university and alterations to the student-university relationship, along with, of course, union dues. Time will...+ adjustments for regional cost of living and inflation) and benefits that were laid out by a 2014 National...

...typically used (e.g., use 5 mL medium on a 10 cm plate). By reducing the culture volume into which the ...transfection and should give rise to a surviving population of approximately 20–50% of the packaging cells...vector using the harvested viral solution as a PCR template. Regardless of the type of virus you’re working..., and allow you to standardize your experiments later. Stay tuned to the Addgene blog and be sure to check...

...barely fits into AAV when you add in a gRNA and regulatory elements. To solve this problem, the Zhang lab...throughout the 23-base sequence. Double mismatches ablated Cpf1 activity, except when they were present in...future edits. In contrast, since Cpf1 cleaves relatively far away from the PAM, NHEJ might retain the ...mechanism. To prevent new editing post-HDR, repair templates should remove the PAM sequence. Many research ...

...blastn), Standard Protein BLAST (blastp), and Translated BLAST (blastx). NCBI has a terrific getting-started...gaps in the nucleotide sequence, you can run a Translated BLAST (blastx) search to check for expected ORFs..., due to the additional searches involved in translating the nucleotide sequence into all six possible...choose for your sequencing result and can easily translate it, we recommend using blastp over blastx. The...

...heartbeat – a muscle contraction – is directly related to the electrical impulse of the cardiac cells....propagated through multiple cells to do things like translate perceived signals to physical actions. Understanding...serotype specificity during delivery (more on this later). Chemigenetic sensors take a hybrid approach and...JF). The voltage-dependent changes in rhodopsin modulate the degree of quenching of the fluor via FRET....

...each gene, we needed to design guide RNAs, donor templates and sequencing primers for quality control. At... taken up to a month to design a single 96-well plate for HDR that satisfy all requirements. To alleviate...guides by CFD score, and it mutates HDR donor templates to avoid re-cutting when needed. It gives you ...the standard information that goes into a 96-well plate and comes out of it. Biotech companies are known...

...of a 5-6 day old human blastocyst [6]. They are isolated mostly from abnormal and/or supernumerary preimplantation...Characteristic ESC ASC Self-renewal High Low once isolated from the human body Usage Universal. Can generate...concern High. Embryos are destroyed during ESC isolation. Low Chances of post-transplantation rejection...when used therapeutically High Low (if ASCs are isolated from the patient) For many years, ESCs or ASCs...

...Thursday, or, at the latest on Friday. It’s best to split cells before the plates are more than 80-85%...Benjamin Franklin)! It’s good practice to label your plates and/or flasks with your name, date, the cell line...disorganization, apoptosis, and gene expression disregulation on HeLa cells." Journal of applied phycology27.6...

...2024). Chromatin context-dependent regulation and epigenetic manipulation of prime editing. Cell. https:/...primer binding site and a reverse transcription template (RTT). The design of the pegRNA has been shown...editing for precise and highly versatile genome manipulation. Nature Reviews Genetics, 24(3), Article 3. ...

...communicate the nuances of protein structure in a flat 2-D images. In the early days of structural biology...full-color images of proteins. But they are still flat. To truly appreciate the 3D structure of these proteins...   He then said “I hadn’t realized how perfectly flat the complex is on this surface”….where the complex...cleavage”. Jacob computationally added the non-template strand to the Jinek structure so that the resulting...

...protein or its role in a disease, researchers often isolate proteins of interest and examine them using biochemical...protein, it's important to validate its function relative to the untagged, wild-type protein. You may, for...the protein of interest and then analyzing the relative colocalization of their fluorescent signals and...One should also be aware of additional problems related to the properties of FPs themselves. These can ...

...institutions recently reported their inability to recapitulate the results of Gao et al 2016 in a letter to...NgAgo DNA guide is inexpensive and can be 5’-phosphorylated in house using a simple PNK reaction or can... the instability of the supposedly alien 5’-phosphorylated ssDNA guides, which might be rapidly degraded... an ssDNA guide. It is possible that, during translation, only a small fraction of nascent NgAgo protein...

...trivia, and everything in between). Eleven years later,  Addgene is a company of fifty instead of three...situation- do we work so hard so we have time to play later? Or, does our play encourage us to work hard? Whatever...employees. The teambuilding culture helps us build relationships and thereby work better together.I realize I...encourage everyone to make an effort to build relationships with their colleagues. It can be easy to get...

...field to a grinding stop and only now, 2 decades later, is it finally back in the limelight. On December...and severe visual impairment that faithfully recapitulated the human condition. Luck brought all these ...X-linked retinitis pigmentosa, achromatopsia, and age-related macular degeneration are ongoing and some will ...capsid variants, designing expression cassettes to regulate and optimize transgene levels, and large scale...

...would convert adenine to inosine, which the translational machinery treats like guanine. This RNA editor...would permit point mutations in RNA which could recapitulate or rescue known pathogenic alleles, or introduce...ADAR will preferentially edit an adenine if the template has a cytosine mismatch at that position. PspCas13b-ADAR2DD...RNA Editing Capabilities through RESCUE Two years later, the Zhang lab expanded the RNA editing toolkit ...

...called CROPseq-multi (Walton et al. 2024). This platform is compatible with enrichment, single-cell sequencing...fast-spiking neurons. The same A-to-D mutation in the related voltage indicator Ace2N-mNeon also led to similar...donor site; SAS, splice acceptor site; pA, polyadenylation signal. Image reused from Riedmayr et al. 2023...therapies. REVeRT supports advanced gene expression regulation, enhancing both the mechanisms of delivery and...

...Regardless, the amount of reporter protein activity correlates to both the amount of DNA transfected into the...general, technical replicates can be thought of as "plate controls". They are NOT independent and are typically...transfection of multiple separate wells (within the same plate) with purified plasmid from the same aliquot/preparation... the same source and were incubated on the same plate at the same time. Although the replicate control...

...to be translated into Modern Greek to be easily accessible today. Roughly speaking, we translated our software...sprints is themed and the stories in that sprint relate to the theme. For example, we could have a shipping...feature to work. We will then go back to this area later and tidy it up. Conclusion While none of us are...

... added on a regular basis, so the ecosystem of related tools is always growing. Beyond the ability to ...formatting. In this way data is moved from a static and isolated vehicle and becomes part of the dynamic and connected...understand and/or attempt to reproduce research relating to data deposited into a repository access to ...can be viewed – and conceivably additional work relating to reproducing the original research can be documented...