...transcript-stabilizing and nuclear export elements such as WPRE (5) and CTE (6), and/or use weaker poly(A) signals like...additional helper sequences (2-3). As alluded to above, termination and polyadenylation of Polymerase II...how the Promoter Controls Gene Expression Learn about New Cloning Techniques Read all of our Plasmids ...

...and G nucleotides). The conversion of cytosine to 5-methylcytosine in CpG islands makes  genes less transcriptionally... the histone protein of interest (in the figure above, H3 at K9 or K27). Joined to this by a flexible ... in Your Experiments Post Read Other Blog Posts about Fluorescent Proteins  ...

...non-lytic insect cell expression systems (sf9, Sf21, Hi-5 cells)  that allow for continuous expression of genes...Resources on the Addgene Blog Learn about inducible promoters Learn about repressible promoters Check out ...

... plasmids bearing one of the distal-less homeobox 5 and 6 (Dlx5/6) enhancer elements and combined it with... and constitutively expressed sgRNA targeting the 5’ end of each essential gene. Koo M, et al. Cell Systems...but be sure to let us know if you'd like to write about your plasmids in a future blog post. No time to ...repository. These variants can be used as tools to learn about the influence of structure on an FP’s properties...100bp or 1kb fragments are produced (see figure above). 3 Reasons The PSU Ladders Are Awesome Tools: ...

...PMID: 28098143. PubMed Central PMCID: PMC5253684. 5. Harrington, Lucas B., et al. "A thermostable Cas9...scientists to make genomic alterations, bringing about a revolution in genome engineering, with new techniques...available by both the Jacob Corn and Alex Schier laboratories. Delivering Cas9-gRNA ribonucleoproteins  An...delivery of RNPs is electroporation (A in the figure above), which generates pores in the cell membrane, allowing...as lipid-mediated transfection (B in the figure above), are in the early stages of development. David ...proteins harboring receptor ligands (C in the figure above), which result in the internalization of Cas9-gRNA...show promise for in vivo studies, little is known about the immunogenicity and stability of the Cas9 protein...

...that may be more tailored to your field of study. 5 great resources to help get your fly work started ...larval stage, Drosophila will pupate, which takes about one week from the initial deposition of the fertilized...there are many resources to help you learn more about fly biology and fly-based experimental systems. ...resources will direct you to the latest information about gene function, while others offer fly stocks and...and other reagents. If you’re ever confused about the best way to study YFG in flies, there is likely an...absolutely essential. You can look up information about your fly gene using the search box in the upper ...In the next post, you’ll learn the fundamentals about how many of these resources work. Many thanks to...

...industry postdoc  Time: Academic postdocs can be long—5 years or more, especially in the biological sciences—whereas...positive experience, I was still incredibly nervous about leaving academia. A lot of academics tend to view...which may take a longer time to acquire. Learning about drug discovery: Unless your academic lab is closely...information. Working as a team: Industry is all about teamwork. A single individual could not bring a ...someone with whom you can have an in-depth discussion about your project might be challenging. In an academic...contacts to get that referral and find out more about the company’s program. Once you’ve found a few positions...

...PMID: 21209361. PubMed Central PMCID: PMC3074624. 5. Bindels, Daphne S., et al. "mScarlet: a bright monomeric...the context of the cell. However, much knowledge about functionality can be gained by understanding the...subcellular marker proteins, it’s possible to get an idea about the subcellular structures your protein resides ...with. These “colocalization” studies give insights about the proximity of two proteins within the same subcellular...Additional Resources on the Addgene Blog Learn More about Plasmids for Endogenous Gene Tagging in Human Cells...

... usually have their comfort zone around 37 °C and 5% CO2 and get agitated with sudden changes in temperature...of the hood while work is in progress.Learn more about cell culture contamination in this post from the... bath constantly at 37°C for this purpose.Learn about the effects of CO2 on sample pH Cell morphology:... Set up a routine to split cells Don’t be lazy about splitting cells, instead try to form a routine. ...secretion of lactic acid (a by-product of cellular metabolism that is toxic to the cells), and need to be split...

...unique single stranded DNA primers that bind upstream (5’) and downstream (3’) of the DNA region of interest...If you have ever worked in a molecular biology laboratory you have likely done a polymerase chain reaction...uses ddPCR for AAV titrating. You can learn all about ddPCR for AAV titration and find helpful tips and...method that can help save time and effort in the laboratory.  There are two main categories of multiplex ...use a single pair of primers, like those stated above. Multiplex PCR is also used in high throughput SNP...

...specific protospacer adjacent motif (PAM) made up of 5′-NGG-3′ (“N” can be any nucleotide base, followed ...more blog posts about base editing Learn about another evolved base editor: ABE8e Read about xCas9, a CRISPR...

...Conditions Rather than Specific Ligands. Eneuro, 6(5), ENEURO.0171-19.2019. https://doi.org/10.1523/ENEURO...posts Read blog posts about neuroscience Resources on addgene.org Learn more about chemogenetics in our...as a DREADD agonist due to its ability to be metabolized into clozapine (CLZ). While some scientists argue...

...human c-myc proto-oncogene product. Mol Cell Biol 5:3610–3616. https://doi.org/10.1128/mcb.5.12.3610-3616.1985...Do you ever wonder about the origins of some of the common techniques or tools you use in the lab? Take...times of the myc tag.  Origins of protein tags For about as long as the disciplines have existed, cell and...examples of new versions of Myc1-9E10 mentioned above basically involve taking the Myc1-9E10 variable ...

... mascot Blugene (check back on December 5 to learn more about this!). I’ve assembled a series of screenshots...search. These pages provide helpful information about the subject, link to related guides and protocols...Additional resources on the Addgene blog Learn more about other features on Addgene's website ...

...challenged with the S. islandicus CRISPR-Cas system. 5. Phage therapy Phage therapy uses phage to treat bacterial...Bondy-Denomy et al., 2018). For example, AcrIIA4 mentioned above indicates that it’s the fourth Acr protein discovered...Cas9 Variant for Your Next Experiment Read more about Optogenetic Control of CRISPR   Resources on Addgene.org...

...substances to laboratory animals: routes of administration and factors to consider. PubMed, 50(5), 600–613...Routes of administration. In H. Hedrich (Ed.), The Laboratory Mouse. Academic Press. Turner, P. V., Brabb, ...

...PMID: 22589270. PubMed Central PMCID: PMC3354961. 5. Sun, Qi, et al. "Discovery of small molecules that... cancer are yielding promising results in the laboratory, and Addgene is accelerating this research by...within that pathway that are of interest to their laboratory. A single mouse click on a gene of interest will...

...client to prioritize investments over the next 3, 5, or 10 years. Complicating these challenges is the...company to collaborate whenever it faces an important challenge. The types of projects mentioned above are just...and why you might enjoy doing it. I’ll also talk about the skills you’ll need to do well as a healthcare...often work with such teams to help make decisions about R&D investments in either new internal efforts, ..., or through acquisitions of or collaborations with other companies. Medical Affairs groups work on building...building awareness among physicians and scientists about clinical research, drugs, the diseases they can ...professionals running the marketing, commercialization, collaborative, and financial functions associated with the...

...with your legal department) right in your breakroom. 5. Throw some physical activity into the mix It’s easy...early employees spent this remaining time by elaborately pranking each other, dressing in costume, and...frequently ask the question, “What’s your favorite part about working at Addgene?” Most Addgenies reply with an... confident that their teammates and project collaborators have completed their work thoroughly. It’s kind...rose-colored-glasses-esque picture of how everyone is kind and collaborative and all we do is have fun all day long, whistling...megaphone, or your company’s intranet. Don’t forget about the power of simple word of mouth - be sure to bring...company.        Additional Resources Read other posts about life at Addgene!Read more of our careers posts! ...

...grants. What is a grant professional?  In the past 5-10 years, the Grants Profession has become just that...on becoming grant writers. When we asked others about how they became grant professionals, mid- to late-career...late-career grant professionals have similar stories about a time when their passion for an organization or... logic models, established partnerships and collaborations, and commitments to and proof of compliance...grants. It is no surprise that the U.S. Bureau of Labor and Statistics shows that employment of grant writers... skills & passion  Skills such as research, collaboration across multiple disciplines, writing, project...development strategy builds on the steps listed above and formalizes your development. You may consider...