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...those entering science communication (myself included) come from a scientific background and bring with... (and I hope I’m not alone in this sentiment). Entering the final semester of a neuroscience major, I ...first semester in which I had several projects nearing deadline (an animated video depicting nuclear fission...level of career satisfaction indeed.   For many venturing forth from tertiary study, the myriad trajectories... scientific research is, after all, one of the enduring goals of science communication.   The Owheo...improve the world around us has long been known; ensuring all have access to this power and the tools to...

... genome engineering, CRISPR is much more efficient at generating the desired mutant offspring, with 85%...adults mated to produce completely transgenic offspring. Although powerful, this technology is cumbersome... Advantages of using CRISPR for mouse genome engineering From the very start, generating mouse models ...microinjectionist, who has the strategically important job of delivering the payload! Last but not least, when using CRISPR...Marraffini LA, Zhang F (2013) Multiplex Genome Engineering Using CRISPR/Cas Systems. Science 339:819–823...Conditional Alleles by CRISPR/Cas-Mediated Genome Engineering. Cell 154:1370–1379 . https://doi.org/10.1016...Multiple Genes by CRISPR/Cas-Mediated Genome Engineering. Cell 153:910–918 . https://doi.org/10.1016/j.cell...

... body recognizes a drug as a foreign antigen, triggering a humoral immune response. ADAs can inactivate...inactivate the drug and increase drug clearance, lowering its efficacy and increasing the risk of an adverse ...on the immunogenicity of the drug product while ensuring the safety and the efficacy.  Better together ...determining the T-cell epitopes that will likely bring about unwanted immunogenicity. The 3-D models can...Monoclonal Antibodies in Macaca fascicularis: From Manufacturing to Preclinical Studies. In J. Bluemel, S. Korte...10.1634/theoncologist.2016-0061WHO. International monitoring of adverse reactions to drugs: adverse reaction...WHO Collaborating Centre for International Drug Monitoring, Uppsala, Sweden, 1992. Mattei, A. E., Gutierrez...

... visiting scientist is distinct from equipment sharing or a collaboration – it requires re-location of...training so that you yourself can become an expert and bring the knowledge back to your home lab. Alternatively...the conversation. Many researchers are open to sharing knowledge and establishing collaborations, so it...if they have specialized skills and equipment! Preparing for your visit   Design and communicate a research...mentor or anyone else available to directly help you during your visit. Reach out to your bench mentor (if ...and rights for anything that could be generated during the visit. If you don’t feel comfortable initiating...travels! Resources Resources on the Addgene blog Fostering communication in remote collaborations Transferable...

... Connie Cepko points out the limited choices in hiring graduate students and postdocs when starting out...counter-productive." He also emphasizes the point to “encourage sharing - in part by having projects ambitious enough that...position, start studying up before you begin the hiring process. Read Genevive Bjorn's "Ready, Set, Hire...Nature Jobs for some quick advice and resources for hiring research staff. And for a more indepth resource...experiments and grant writing. You have to enjoy the mentoring role.” She points out that new PIs may not recognize...student or postdoc may require different types of mentoring. Some mentees prefer lots of advice and micromanagement... interdisciplinary, with emphasis on genetic engineering, sequencing, and translation of results to companies...

...world of academic research can be quite daunting. During my first year of college, I knew I wanted to try... four years after I started college, I will be entering a PhD program to continue my scientific endeavors...met graduate students who enjoy the thrill of acquiring a piece of knowledge before anyone else in the...neurodegenerative disease because they had close relatives suffering from similar diseases. Others were inspired by...biofuel production. There’s a lot out there worth exploring! Being excited about the science allows you to...proactive Rather than applying only to posted job offerings, the best research experiences typically come ... provided me with much-needed emotional support during my journey. When you are hunting for labs to join...

...take a look at a few of these methods below. Engineering gut microbes in situ While the ability to monitor... University developed a method for genetically altering gut microbiome members in their native habitat...reporter gene. The hairpin blocks translation by sequestering the RBS, but binding of the “trigger RNA” (in... Counting cell division in the gut microbiota Monitoring microbial population dynamics in the gut can ...one bright particle that segregates into one cell during cell division. To achieve the agglomeration of ...References Leonard, Sean P., et al. "Genetic engineering of bee gut microbiome bacteria with a toolkit...PMC4674677. Ronda, Carlotta, et al. "Metagenomic engineering of the mammalian gut microbiome in situ." Nature...

...allow authors to avoid many of these issues, by sharing their work publicly and immediately once the manuscript...emerging COVID-19 pandemic quickly and openly. By sharing results faster and more broadly, scientists can...evidence of their productivity that can be shared with hiring and review committees, as opposed to the opaque.... Preprints at Addgene Preprints are all about sharing scientific knowledge faster and more openly — which...open-science organizations, we support scientists in sharing their results and materials with the broader community...in a preprint provides all the same benefits as sharing those plasmids alongside a journal article, but...He also posted two preprints of his own research during his PhD studies.  References and Additional Resources...

... neurons using GCaMP7, measuring calcium signaling related fluorescence during morphine administration... neurons at specific firing frequencies. She hopes to identify how in vivo firing frequency of PV neurons...researchers attend a conference of their choice while honoring the late Michael Davidson and Roger Tsien who ...iGluSnFR3 (left). GluSnFR3 in pMinDisplay (PDGFR anchoring domain), in electroporated in rat hippocampal ...modulate protein activity and have important roles during oxidative stress. To monitor this activity, Pedre...

... genetic engineering, with the ultimate objective of genome design.” It’s genetic engineering on a genomic...toolbox needed for anyone to begin considering genetic engineering projects in a very big way. In one ...scale could be created. Someone could take on an engineering project including hundreds of genes and regulation...complete toolbox for taking on very large genetic engineering projects. Addgene: How easy is it to do this ...animals make. That will be an outcome of genetic engineering.   Thank you to Chris Voigt for speaking with...

...2023). Uncovering the functional diversity of rare CRISPR-Cas systems with deep terascale clustering. Science...interest in bacterial and archaeal immune systems. During this time, scientists discovered many related immune...keep increasing, as efforts using large-scale clustering and discovery algorithms find new candidates ... L. A., & Zhang, F. (2013). Multiplex genome engineering using CRISPR/CAS systems. Science, 339(6121),...1027. https://doi.org/10.1126/science.aaq0180 Harrington, L. B., Burstein, D., Chen, J. S., Paez-Espino..., K. V. (2021). Type III CRISPR-CAS systems: deciphering the most complex prokaryotic immune system. Biochemistry...(2015). Co-transcriptional DNA and RNA Cleavage during Type III CRISPR-Cas Immunity. Cell, 161(5), 1164...

...CRY2, although some studies have shown that this clustering only happens under certain conditions. With...drawback of this method is the stray excitation of neighboring neurons in opsin dense regions. To overcome this...able to demonstrate that while illumination of neighboring cells evokes action potentials (APs) in patched...this operon can serve as a living biosensor for measuring caffeine content in samples. The Barrick lab and...concentrations in several beverages could be estimated by measuring growth of the ΔguaB pDCAF3 strain. Find the decaffeination...placesTagRFP-T and Dronpa in close proximity. By monitoring changes in TagRFP-T fluorescence intensity at...the authors tracked protein activity while also capturing information about spatial and temporal organization...

...population. Outbred strains of mice are useful for comparing dominant and recessive traits and their impact...or treatments, but are not useful for genetic engineering. Examples of outbred strains include Swiss Webster...generating knockin mice but because CRISPR/Cas9 engineering favors DNA repair via non-homologous end joining...have errors introduced to the nucleotide sequence during the DNA repair process. The addition of small molecules...to transgenic mice, knockout mice are engineered during the embryonic stem cell stage using one of several...blog posts about using Cre-lox Read our genome engineering blog posts Resources on Addgene.org Find more...more about Cre-lox Read our Genome Engineering Guide Learn more in our CRIPSR guide ...

... while engineering viruses Acr genes can be used as positive selectable markers in engineering viruses...Acr proteins can also be used to prevent dCas9 tethering to DNA. Since these early anti-CRISPR studies,...dimerize which blocks the PAM recognition site (Harrington et al., 2017). Inhibit crRNA loadingExample: ...domain of Cas9 and prevents target DNA cleavage (Harrington et al., 2017). Figure 1: Anti-CRISPR proteins...436–439 . https://doi.org/10.1038/nature22377 Harrington LB, Doxzen KW, Ma E, Liu J-J, Knott GJ, Edraki... EJ, Maxwell KL, Davidson AR (2016) Naturally Occurring Off-Switches for CRISPR-Cas9. Cell 167:1829-1838...

...guide sequence for your genome engineering experiment. With CRISPR entering the clinic for correction of...structure-guided engineering of the PAM-interacting domain of Cas9. After multiple rounds of engineering, they developed...PAM right where you need it, the odds of this occurring are not in your favor. Thus, there is a need to...coverage in the human genome. SpRY Cas9 has PAM sites covering the majority of the genome! Want to get your...Requirement and Expanding Beyond Cas9 xCas9: Engineering a CRISPR Variant with PAM Flexibility History...

... Cold Spring Harbor Protocols. 2012 Jun 1;2012(6):pdb-top069609. Palmer AE, Tsien RY. Measuring calcium...concentrations. This will reduce noise and ensure you are measuring the events-of-interest. Sensors suitable for ER...reporters use a single fluorescent protein (FP), requiring only a simple measurement. However, single wavelength...sites per sensor (to decrease unnecessary calcium buffering and to increase sensitivity), and is a great FRET...expression and integration within the target cells, ensuring more consistent and reproducible results in calcium...subtypes for efficient expression and downstream monitoring. This targeted approach allows scientists to ...

...In order to understand the complex wiring of the brain and the underpinnings of neurological disease,...manipulating cell activity, especially neuronal firing. What is chemogenetics? Chemogenetics relies on...looking at immediate effects of specific neuron firing are better equipped using fast-acting optogenetic...receptors can be used to alter excitability by lowering or raising the threshold for signaling. The two...for neuronal signaling and causes neuronal burst firing. Other DREADDs, such as the KORD receptor, increase...The Power of the Lock, the Weakness of the Key. Favoring the Pursuit of Specific Conditions Rather than...Carpenter JC, Snowball A, Knauss S, von Schimmelmann M, During MJ, Lignani G, Schorge S, Young D, Kullmann DM,...

...amount of time. Though I now spend far less time answering help emails and phone calls, I feel compelled ...can be a particularly good resource if you are considering using a reagent for “off-label” purposes. When... than you might think. The Addgene scientists answering the phone are smart, but they cannot (and should..., but many bio companies have real scientists answering their technical support questions (besides Addgene...Addgene, I know both Abcam and NEB have Ph.D.s answering emails and phones). We can’t be experts on every...-up when things get crazy. Take home message Answering customer service questions is a delicate balance...Once You Recieve Them Get Help with the Addgene Ordering Process ...

..., the vector may undergo genomic rearrangements during replication. To avoid genomic rearrangements, try...store freshly harvested virus at 4 °C instead of storing it briefly at -80 °C. If troubleshooting, the safest...increased by concentrating a viral stock. Different titering methods can reveal differences in physical versus...result of plasmid integration via recombination during transfection, and does not necessarily indicate... infect the packaging cells. Another method of ensuring viral production is to perform PCR against components...adoptive immunotherapy." Acute Leukemias VIII. Springer Berlin Heidelberg, 2001. 291-306. PubMed PMID:...Yezzi, and Barbara R. Young. "Recombination events during integration of transfected DNA into normal human...

... important consideration when preparing the implant itself, which brings me to the second key point; the...cable to the ferrule implant must be maintained during the entire protocol, otherwise optical stimulation...it to a patch cable connected to the laser and measuring its output with a light meter before and after...that one laser can stimulate only a single animal during the course of an experiment. There are a number..., 6]. These have the obvious advantage of not requiring patch cable connection to the laser, which affords...science writing. Derek is also a pianist and an aspiring composer. View his blog drsimonsaysscience.org...