...repression, high precision edits, and more. Figure 3: Schematic of the basic CRISPR-Cas9 mechanism, representing...double-stranded breaks caused by Cas12 create short 3’ overhangs, which leads to slightly increased efficiency...for genome-editing applications. Molecular Cell, 84(3), 463-475.e5. https://doi.org/10.1016/j.molcel.2023.12.034...the type I CRISPR system. Gene and Genome Editing, 3–4, 100013. https://doi.org/10.1016/j.ggedit.2022.100013...Class 1 is the most abundant of the two, comprising about 90% of identified CRISPR-Cas in bacteria and nearly...CRISPR systems called CasPEDIA, with information about enzyme activity, experimental conditions, and more...

...can also find Dr. Knoepfler on Twitter: @pknoepfle 3. "Social Media as a Scientist: A Very Quick Guide"...tips for using various social media platforms, and 3) helped you fit social media into your busy schedule... advantage. It wasn’t until I wrote a blog post about a research paper that explored how scientists connect...saw a request for an Addgene guest blogger post about colony PCR, I signed up immediately. I had used ...science communication skills, while also learning about new scientific developments and helping scientists... found this paper. I wrote an Addgene blog post about it and shared the paper with my lab since one of...project. I was invited to participate on a panel about “Social Media for Science Communication” at the ...

... near-infrared photosensitizer. Nature Methods,13(3), 263-268. doi:10.1038/nmeth.3735. PubMed PMID: 26808669...PMID: 18706812. PubMed Central PMCID: PMC4445427. 3. Bulina, M. E., Chudakov, D. M., Britanova, O. V.,... in culture and in vivo.  Read on to learn more about this killer illumination technique! What is a photosensitizer...within the near-infrared (NIR) spectrum (>620 nm). Above is a brief schematic of how the MG-2I-dL5** FAP-...use MG-2I-dL5** with your research? Let us know about your experiences using photosensitizers in the comments... to Receive the Fluorescent Protein eBook Learn about Small Molecule Fluorophores Resources on Addgene.org...

...MS2 RNA aptamers at the tetraloop and stem-loop 2; 3. The MS2-P65-HSF1 plasmid which expresses the activation...LS, Kampmann M, Weissman JS. Cell. 2014 Oct 23;159(3):647-61. doi: 10.1016/j.cell.2014.09.029. Epub 2014...that screen with a system you are really confident about working for most genes and individual targeting ... system, Konermann explained, was new knowledge about Cas9’s crystal structure. “I thought maybe we could...RNA we use. What we have to do now is learn more about the rules.” He said that there is no theoretical...References & Further Reading: For more information about SAM, including a tool for getting optimized guide...

...random-walk" of a budding startup, I asked Mike to outline 3 pieces that are essential to the biotech startup puzzle... anywhere. The team should initially consist of 2-3 individuals that are aligned in vision and outcomes...transitioned away from the bench and became all about finding the best product-market fit for this set...Notably, the team must have an upfront conversation about its goals and vision for the company; this will ...section! And read on for more great information about starting a biotech company. Suggested reading The...of Sample6) for taking the time to speak with us about the biotech startup process and work environment...

... Our Segment on these new AAV Vectors Hot Plasmid 3: mCyRFP1 - a new, monomeric, and cyan-excitable red...of our Hot Plasmids podcast series, you'll learn about new red fluorescent proteins, AAV tools for targeting...

...segment on modulating iPSC reprogramming  Hot Plasmid 3: Regulating telomere length with TZAP. Listen to the...of our Hot Plasmids podcast series, you'll learn about new libraries for studying B. subtilis biology, ...

...to the Segement on Ouabain Selection  Hot Plasmid 3: CRISPR base editors with enhanced range and specificity...of our Hot Plasmids podcast series, you'll learn about optogenetics tools for controlling protein activity...

... cover letter can be found on the Addgene blog.   3. Personal statement brilliance In addition to sending... Letters of recommendation wisdom  Reach out to 2-3 individuals in your professional circle to vouch for...Rose is passionate about outreach among low-income communities to educate parents about resources that can...automated filters of consideration. If you are worried about any eligibility provision, don’t guess. Simply reach...

...modified from pSC101 with copy numbers ranging from 3-100 per cell, a stabilized promoter buffered against... expression levels returned to the baseline after 3 hours with the stabilized promoter while gene expression...is useful for many metabolic engineering and synthetic biology goals. In metabolic engineering, the expression...or can interfere with the precision needed in metabolic engineering or other synthetic biology circuits...

... they can deplete endogenous protein levels for 1-3 days. There’s also a CRISPR for that! CRISPR interference...most of the other mechanisms described. Fig. 3 dTag mechanism of protein degradation. Adapted from...which can be used with this approach - read more about them on our CRISPRa blog! Fig. 1 CRISPR ... is a lot of overlap with the systems discussed above, just with more elements of control. Tet promoters...to be site-specifically knocked in. This can be labor intensive and time consuming. If your native protein...

...“blind” to this very important difference (Figure 3). If you are interested in harnessing the power ... PubMed PMID: 27089539. PubMed PMCID: PMC4835065. 3. Mock, U., Hauber, I. & Fehse, B. Digital PCR to assess...advantages over mismatch nuclease assays that will be elaborated below. Digital PCR Digital PCR (dPCR) involves...type targets. Comparatively, detection limits of about 5% mutant targets have been reported for mismatch...

...provide problem-solving platforms.     Fig. 3: Different workflows involved in sharing open hardware... (2022). https://doi.org/10.1038/s41592-022-01602-3... in science brings stronger networking across laboratory and disciplinary boundaries and can allow for... that are otherwise cut off from the process.   About the author: Benedict Diederich received his PhD ...

...no addition of nucleotides to the 5’ spacers. The 3’ ends were modified slightly with either a 1-7 base...constructs, transcript levels of PTG1 and PTG2 were 3 and 31 times higher, respectively. In rice protoplasts...with limited packaging capacity. Still confused about the various tRNA expression strategies? No worries... Addgene. If you have any questions or thoughts about this technology, leave them in the comments below...

... of conventional gene targeting is lowered to about 2-3 months. But certain models remain more difficult...directly into a zygote and obtain a knockout mouse in about 3 months. Image courtesy of Wenning Qin. Single-stranded...cleavage right before the stop codon!  Figure 3: How to design a C-terminal insertion with Easi-CRISPR... on CRISPR Mouse Models Learn about Homology Directed Repair Learn about Cas9 Ribonucleoprotein (RNP) ... a variety of species, including the standard laboratory mouse. It’s also made targeted insertions relatively... a mouse embryo to create chimeric mice. This laborious process takes ~8-10 months. With CRISPR (B), you...

...drug clinical trial, for example).   Fig. 3: Question flowchart for choosing an ANOVA/MANOVA statistical...positive is much higher than if you only look at 2 or 3 pairwise comparisons. Similar to family-wise error...for the nonstatistician: Part I. South Med J., 105(3): 126-130 (2012). 10.1097/SMJ.0b013e3182498ad5 Curran-Everett...a summary snapshot without telling you anything about outliers or data spread within those data sets. ...the rules” and which we have certain information about. The assumptions and requirements that need to be...Quantitative If your data doesn’t fit the requirements above, a non-parametric test may be the way to go, which...only performing the comparisons you really care about. Family-wise error is the probability of a false...

...Ten Simple Rules for a Successful Collaboration. PLoS Comput Biol 3:e44 . https://doi.org/10.1371/journal.pcbi...else to focus on. Use collaborative cloud apps It is a good idea to use collaborative cloud tools such as...creative collaboration in the workplace. There are many other ways to encourage collaborations remotely...meeting? No problem! You can still creatively collaborate and brainstorm! Image from Joseph Mucira. ...serve as a springboard for future discussion. Collaboration and creativity are pivotal in writing publications...Vicens et al., 2007) geared towards enhancing collaborative writing and this recent article published in...approaches/tools we have to foster creative collaboration remotely in science are endless. Let’s adapt...

...2015 Dec 31. pii: aad5725. PubMed PMID: 26721683. 3. Nelson CE, Hakim CH, Ousterout DG, Thakore PI, Moreb... some research in human embryos. Nature. 2015 Dec 3. doi:10.1038/nature.2015.18947 5. Siva K, Covello ... by zinc finger nucleases. Mol Ther. 2015a Mar;23(3):523-32. PubMed PMID: 25492562. PubMed Central PMCID...estimated that very little dystrophin correction (about 4%) is needed to see muscle improvement, with only...diseases we’d like to treat with CRISPR. As seen above, DMD can be treated with “one-size-fits-most” NHEJ-mediated.... Precision genome editing faces the challenges above and more. One chief challenge is upregulating HDR...

... the pH change caused by storage on dry ice (Fig. 3). However, 30 seconds was not sufficient to prevent... was a substantial reduction in infectivity after 3 weeks of exposure to pH 5.1 (Potter et al., 2014)....return the sample back to the original pH. What about putting it back in the freezer? We then tested whether...

...break membranes outside of their natural context or 3) be modified to extend at different pH levels. Domesticating...Protistology 48, 290–296 (2012). PubMed PMID: 22356923. 3. Polka, J. K. & Silver, P. A. A Tunable Protein Piston... Additional Resources on the Addgene Blog Read About Other Hot Plasmid Technologies Read Other Synthetic...