...2010 Yuhai Cui Red mScarlet Split-wrmScarlet variant of mScarlet Split-wrmScarlet and split-sfGFP: tools...

...thereby undo the genetic changes driven by the earlier intervention2. While an initial reversal drive ... responsibly. An accidental release would be particularly dangerous for molecular biology and genetics...aaa5945 (2015). doi:10.1126/science.aaa5945. PubMed. DiCarlo, J. E., Chavez, A., Dietz, S. L., Esvelt, K. M....013896 (2015). doi:10.1101/013896. bioRxiv. Fitz-Earle, M., Holm, D. G. & Suzuki, D. T. Genetic Control...

...reef-building corals (Dove et al., 1995). In the early 2000s, a handful of chromoproteins were identified...ShadowR is used as a FRET acceptor to mRuby2 or mScarlet. Image: Murakoshi et al., 2019. ShadowR is ... a FRET acceptor in combination with mRuby2 or mScarlet as the donor in HeLa cells.   Find ShadowR for... New Suite of Allelic-Exchange Vectors for the Scarless Modification of Proteobacterial Genomes. Applied...

...cloning approach should you use if the budget is particularly tight? Most cloning processes will require a... their final order through sequence homology. Similarly, Golden Gate assembly allows one to simultaneously...simultaneously complete an ordered set of scarless ligations following digestion with Type II restriction enzymes... options could be good choices if you have a particularly large number of inserts to assemble. You could...

... HEK and CHO do not seem to require this, while nearly all endocrine cells do. While not all of the wells...in groups as they become confluent, taking both early- and late-arising clones, and making sure we always...Lindberg papers in PubMed). While we found that nearly all of the cell lines we tested were amenable to...Maryland Medical School in Baltimore. She is particularly interested in signaling peptide maturation in...

...over time! Communicate often and clearly As a manager you need to clearly state expectations and goals. ...yourself anymore. Personally I found this to be particularly challenging. As PhD students and postdocs we...there is not a clear deadline. Finally check-in regularly, make yourself available, and encourage questions...

... the other person seemed confused, bored, or particularly excited about? If you tried tweaking your pitch...During the conference Arrive at the poster session early to get your poster up on the board and meet your... contact information if your conversation is particularly fruitful. After the conference Bring back what...LinkedIn or Twitter, for instance. If you’ve had a particularly meaningful conversation, and you have their ...

... attempting knock-ins by HDR.   Fig. 1: Early steps in repair of a DNA double strand break by ...Promoting HDR events over other edits Like I said earlier, HDR is not the most common editing pathway for...generating ssDNA overhangs at the break site – an early HR step (Moreno-Mateos, et al., Nat Com). Another...for HR is to fuse an HR protein to the nuclease. Early HR factors fused to Cas9 have been shown to increase...

... becoming increasingly politicized. It would be nearly impossible for a career scientist such as Zimmer...up in this book. Some of these issues are not particularly new. Chapter 2, “The Professional Scientist,...and seeing names like Feng Zhang, Jennifer Doudna, Karl Deisseroth, and Ed Boyden crop up in this book was... process and the marginalization of science, particularly public health and environmental science, by ...

... a gating strategy from a published experiment (Barlow-Anacker et al., 2017). This gating strategy is ...        Figure 4: Gating strategy used in Barlow-Anacker et al., 2017 to identify mDCs and pDCs....      Figure 5: The gating strategy from Barlow-Anacker et al., 2017, with quadrants to help visualize..., 53–59. https://doi.org/10.6026/97320630014053 Barlow-Anacker, A., Bochkov, Y., Gern, J., & Seroogy, ...

...prior to running my flow analysis, I can however clearly identify my lymphocyte population (10.7% of all...my negative, unstained population, which I can clearly differentiate from the positive one (Figure 8).... that in many cases a stain will not result in clearly distinct positive and negative populations. Oftentimes...Paul Heisig is a Research Associate in the lab of Arlene Sharpe at Harvard Medical School. His projects ...

...good idea to start thinking about public speaking early and often. Two of my jobs have indirectly resulted...learn from. Some of the best talks I have seen by early career scientists are those in the Young Scientist...appropriately.  Find out who will be listening.  Are they early, mid, or late career?  Will many non-scientists ...make it suspenseful. 4. Start preparing your talk early - like today Don’t wait until you need to speak ...

...is transferred to the acceptor fluorophore, a circularly permuted Venus. In the case of pHlash, the signal...fusion proteins or tags and reporter genes can be nearly impossible. Fortunately, a collection of plasmids...applications of CRISPR/Cas9 in genome-engineering. One particularly attractive application involves using dCas9-... The easily reusable MXS-building blocks are particularly effective at generating plasmids with varying...them, fused them to GFP, and compared them to popularly used epitopes tags such as HA and MYC in vivo.... ER) Microtubules mCherry-alpha-tubulin Early Endosome Vacuolar Compartment mCherry-Rab5 BFP-Rab5...based genome editing techniques out there (and particularly CRISPR), it is now relatively easy to design...

...interrogate the activity of uncharacterized promoters. Similarly, placing UAS upstream of a transgene permits directed... binary systems above if they all behave very similarly?” The simple answer is that sometimes it is necessary...trans-activator of herpes simplex virus immediate early gene expression. Genes Dev. 1988 Jun;2(6):718-29...1472-6750-6-43Kaczmarczyk A, Vorholt JA, Francez-Charlot A. Cumate-inducible gene expression system for ...

...PRESTO-TANGO: An open-source resource for drug discovery Nearly one third of all medications target G-protein coupled...altering gene expression are relatively well known, particularly in bacteria, the process of cloning, combining...second-generation CRISPR lentiviral library targets nearly all human protein-coding genes, and is unique in...Vectors Available at Addgene: Lab Plasmids Dan Carlson RCIscript-GoldyTALEN, pC-GoldyTALEN Tom Ellis...Takashi Yamamoto pcDNA-TAL-NC2, pCAGGS-TAL-NC2 Charles Gersbach pcDNA3.1-GoldenGate, pcDNA3.1-GoldenGate-VP64...diverse antibodies; in the CRISPR-X system, AID*Δ similarly produces single base changes at each gRNA-specified...

... from Stanford University, led by neurobiologist Karl Deisseroth, combined optical and genetic methods...30503173. PubMed Central PMCID: PMC6317997. Deisseroth, Karl, et al. "Next-generation optical technologies for...17035522. Pubmed Central PMCID: PMC2820367. Deisseroth, Karl. "Optogenetics." Nature methods 8.1 (2011): 26. ...PMCID: PMC3979449. Fenno, Lief, Ofer Yizhar, and Karl Deisseroth. "The development and application of ...

...masks. Make sure to change the gloves and masks regularly instead of keeping them in the pockets of your...cross-contamination. CO2 incubators should be cleaned regularly and dampness of any kind should be avoided. Distilled...provided to maintain the humidity should be changed regularly as mycoplasma, bacteria, and fungus can reside...and other sources of infection must be cleaned regularly and checked for contamination periodically. For...

...National Science Policy Network to connect with early career science policy groups, attend webinars, and...programs and fellowships have a strong focus on early career researchers who have decided to transition...mentoring others (see more in Figure 2). Starting early demonstrates not only your interest to future positions... impacts of health services research. She is particularly interested in driving scientific and technological...

...school’s equivalent of a senior thesis), I knew early on that I wanted to work with a partner. WPI’s emphasis...would launch our careers in science. Fast forward nearly 20 years and I am known by some colleagues as the...success are defined by ongoing mission fulfillment. Earlier this year, I joined Addgene’s Green Team, a like-minded...post-doc adviser almost never had lab meetings. In my nearly 4 years in the lab, we probably didn’t have more...

...contrast to the acute immune responses that eliminate early stage cancer cells, chronic inflammatory responses...types of immune cells. Dendritic cells (DCs) are particularly popular whole cell therapeutics because they...clinical trials, expansion of participants based on early results can enhance the positive effects of the ...cell deaths that help activate tumor responses. Similarly, some radiotherapies have caused increased death...