...engineering realm of S. cerevisiae and E. coli. The utility of simultaneous and multidirectional multiplex ...off switch The multiple scRNAs all individually utilize the same dCas9 protein. By using an inducible promoter... By using an orthogonal dCas9 protein that will still recognize guide RNA but with a different dCas9 binding...

...neuroengineering, and some of the optogenetic tools that are still missing. Optogenetic tools for neuroengineering ...focus on the next round. Addgene: What tools are still missing from the optogenetics toolbox? Boyden: That's...question. We've made a lot of progress. I would say we still would like to find ways to make these tools more...

...played enormous roles in enabling researchers to utilize and develop these tools by making their own fluorescent...loops on either side of an enzyme’s active site and still block enzyme function upon exposure to ultraviolet...Mek2, Cdk5, and Cas9. Xin has demonstrated the utility of these FLiPs in a variety of research applications...

... That papers with plasmids deposited at Addgene still accrue on average 2 to 3 times as many citations...step. There are lots of other factors that could still be controlled for. As an example, plasmid-related...of correlation. But even with that caveat, it is still suggestive that we see such a big difference in ...

...learning about biology in high school, it wasn’t until I started working in my first molecular biology ... like taking an engine out of the car – you can still use the engine as a power source, but now you don...pellets (FD-CF pellets), leaving them shelf-stable until water is added and protein production proceeds. ...

... alcohol or acetone drop by drop onto the smear until the alcohol runs clear and immediately rinse. Flood...for 30 - 60 seconds. Rinse the slide in tap water until the runoff is clear, blot gently to dry, and then...by the interaction of CV+ and I- in the thick multilayered membrane of the gram positive bacteria. In a...

...limitation. The procedure will also need to be perfected until the efficiency of repair approaches 100 percent.... fundamental questions about CRISPR/Cas biology still left to answer as well, she says, and those details...increasing effectiveness of CRISPR/Cas9, scientists are still looking for ways to diminish the possibility of ...

...requirements (Figure 1), but Cas12a has generally been underutilized in genetic screens because of its lowered editing...coexpressed proteins run as ladders. Image from Santilli, et al. 2021 Plasmids are used to express ...via engineered immunoglobulin binding domains.  Santilli, R.T., et al. Sci Rep (2021). https://doi.org/...

...community-driven life of its own. Nevertheless, there are still many open questions to be clarified, especially ... put their work on the shoulders of giants. The still relatively young movement of sharing open hardware...Diederich, B., Lachmann, R., Carlstedt, S. et al. A versatile and customizable low-cost 3D-printed open standard...

...homologous recombination. To further increase the utility of CRISPR/Cas9, it will be necessary to improve...multiplexing in systems with limited packaging capacity. Still confused about the various tRNA expression strategies...multiplexing afforded by PTGs will improve the utility of CRISPR/Cas9 in new avenues of research. The ...

...Unfortunately, using proteins as bait has limited utility for identifying novel RNA-binding proteins that...BASU, is derived from a BirA* homolog found in B. subtilis. BASU was  compared head-to-head with the E. coli...time point, but one minute of labeling with BASU still generated a >30-fold enrichment compared to E. coli...

..., broad utility for degrading targets in different cellular compartments, and in vivo utility. dTAG molecules...genetic approaches. In addition, the dTAG system is versatile and can be used in both cell-based assays and ...

...however, have a higher conductance, so they can still depolarize neurons at lower MOIs. Figure ...located in the midbrain, fiber-optic cannulas were still stereotaxically implanted into the mice’s brains.... Excitation of neurons deeper in the brain may still require fiber-optic cannula implantation, but altering...

...done significant work to improve the Adv genome’s utility in research and its therapeutic potential. Note...use of this system. New and improved systems are still being developed. A detailed protocol for production...should be used as an internal control (ATCC#VR-1516) until the assay is validated. Infectious titer: (Plaque...

...identifying a suitable route of administration. However, until recently safe delivery of nucleic acid cargo (DNA... of blindness and was thought to be untreatable until 2008. Following a single dose of subretinal administration... and other ongoing clinical trials support the utility of AAV as a broadly applicable gene delivery tool...

...effects. Luckily for us, CRISPR is an incredibly versatile system that can be tweaked to target ncRNA. CRISPR...and its role in gene expression, though this may still unexpectedly affect nearby genes (Figure 1A-C). ...lncRNAs. The Cas14 variant is a smaller and more versatile option. It can be used to target lncRNAs with ...

...focus on one such strategy--the FLEx switch--which utilizes recombination elements to turn off expression ...introduced, recombination can proceed either by first utilizing the loxP sites or the lox551 sites. Regardless...reporter would ensure that the reporter remains silent until a cell or tissue-specific Cre is provided. Since...

...” identified seven types of difficult people: Hostile-aggressives, who are usually antagonistic and impolite...is a difficult person, the above strategies will still be helpful but you will need to be more assertive... try to intimidate you with negative comments (hostile-aggressives, complainers, know-it all experts, ...

...frequency. Despite the progress we’ve made, there is still a need for and interest in enzymes with shorter ...editing; for known promiscuous gRNAs, xCas9 3.7 still showed a 4.2-9.4-fold improvement in off-target:...these early data are exciting, there’s a lot we still don’t know about xCas9. “I’m not 100% sure xCas9...

...after a semester while the other student stayed until he graduated. Years later I attended a formal training...team members by giving them new opportunities. I still need to check myself sometimes to make sure I am.... In many cases reports are not aware of issues until you bring them up. I wouldn’t go as far as saying...