... vectors through pseudotyping.” Curr. Gene. Ther. 5 (2005) 387-398. PubMed PMID: 16101513. PubMed Central...applications to help you start (safely!) thinking about the possibilities. Ready to plan your next (or maybe...S, Varmus H. Retroviruses. Cold Spring Harbor Laboratory Press; Cold Spring Harbor, NY, USA: 1997. PubMed...Additional Resources on the Addgene Blog Learn More about Lentivirus Listen to Connie Cepko Discuss Gene Therapy...Addgene.org Browse Our Viral Vector Pages Learn About Viral Vector Biosafety Deposit Your Viral Vectors...

...Ishitani R, Zhang F, Nureki O. Cell. 2014 Feb 27;156(5):935-49. doi: 10.1016/j.cell.2014.02.001. 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...

...insert. If the insert is large, we may only check the 5’ and 3’ ends to verify that the sequence assembly ...One of the best things about sharing plasmids through Addgene is that we provide an added level of confidence...bp of clean sequence from a typical reaction, or about 250 amino acids worth, so make sure the feature ...96-well microplates (identical copies, see image above). We then select forward and reverse primers for...specifically to target this region, as outlined above. Avoid primers that bind within or must read through...

...Retroviruses and cancer. Rev Infect Dis. 1982 Sep-Oct;4(5):1015-25. DOI: 10.1093/clinids/4.5.1015 Lee C.S., ...clinical settings. AAV is also a cool tool in the laboratory! It is ideal for whole animal administration,... be helpful to you. Interested in learning more about a specific virus? We have you covered with guides..., adenovirus, and AAV. If you’re wondering more about what applications these viruses are good for, we...

...will end up with a stellar letter of support.        5. Application submission  Set a calendar reminder 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...

...phase, the number of plasmids per cell increases 4-5 fold (for pUC plasmids) or ~2 fold (for p15a, R6K,...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...

...PMID: 25775042. PubMed Central PMCID: PMC4414919. 5. Yarmolinsky, Michael B., and L. Gabriel. "Inhibition... FUNCAT Professor Schuman’s technique applies metabolic labeling using a noncanonical amino acid, azidohomoalanine...both quantitative and spatio-temporal information about the turnover of specific endogenous proteins in ... interested in neurobiology.  She is passionate about communicating science to all audiences. Right now...

...degradation. Nature Chemical Biology. May 2018. 14(5): 431-441. PubMed PMID: 29581585. Additional resources...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...

...for your species. Affinity purification Only up to 5% of the IgG antibodies produced by an animal are specific...  Read about another common type of antibody for research, monoclonal antibodies Learn about the advantages...Once the initial antibody response starts to wane about a month later, the animal is given additional booster...

...tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain. PLoS One. 2013 Sep...Resources on the Addgene Blog Learn More About Lentivirus Learn more About AAV Get the Scoop on Our Viral Service...

... are safe! Colony screening Pick a few colonies (5-10) and grow them in a small culture volume (2 or ...they get old, they lose competency). If all the above is correct, you should probably have a second and...blog posts Read blog posts on plasmid cloning Read about different plasmid elements Resources on Addgene.org...

...mechanism, but their small packaging capacity of 4-5 kb limits the delivery of larger transgenes, such ... plasmids you need. If you'd ever like to write about a recent plasmid deposit please sign up here.    ...tracing using the transgenic zebrafish model, ubi:zebrabow. In this system, cells express various fluorescent...

...were never stored on dry ice/exposed to CO2 (Fig. 5). Basically, the CO2 goes in and the CO2 goes out....return the sample back to the original pH. What about putting it back in the freezer? We then tested whether...

...as well as their orientation relative to the gRNA (5’ or 3’). In some cases, there is no requirement, which...you are a structure junky who just wants to learn about Cas domains. CasPEDIA either directly has this information...

...DNA damage. Far-red light, however, can penetrate 5 mm past the surface of skin and is less phototoxic...633–639 . https://doi.org/10.1038/s41592-019-0436-5 Mathony J, Hoffmann MD, Niopek D (2020) Optogenetics...of the genome editing process. Read on to learn about the various light-controlled CRISPR tools available...reaching the amber stop codon, TAG. (To learn more about site-specific incorporation of amino acids using...

...Synth Biol 2014. Feb 5 (Web); DOI: 10.1021/sb4001504. For further reading about Nicola Patron's MoClo.... Steinfeld et al., ACS Chem Biol 2014 May 16;9(5):1104-12. Heinemann et al., FEBS Lett 2012 Oct 19...creating a single transcriptional unit (Ex: a promoter, 5’UTR, coding region, and terminator). Next, up to six...2012 Mar 20;109(12):E690-7. For further reading about SpyLigase technology, read the Addgene interview...in optimizing TALEN assembly and activity? The laboratory of Takashi Yamamoto has created a complete TALEN...pGreen-IIS based destination vectors. To learn more about the GreenGate cloning system, see the detailed plasmid...

...administered at a total dose between 1 x 1011 and 5 x 1011 vg per animal (i.e., 10 and 50 µL of vector...across the Blood-Brain Barrier. Molecular Therapy, 27(5), 912–921. https://doi.org/10.1016/j.ymthe.2019.02.013...targeting distinct brain cell types. Nature Methods, 17(5), 541–550. https://doi.org/10.1038/s41592-020-0799... for their use. Then, we’ll provide information about commonly used and emerging capsids to help you decide...our Caltech Systemic Capsids page to learn more about that collection. Table 1: Systemic capsid plasmids...slices Several of the systemic capsids mentioned above have also been tested on ex vivo brain slices obtained...

...situations where you learn biology terms (plasmid, 5-prime, CRISPR, growth strains, vector backbone, etc...you access to hard data about what steps are most likely to fail. Our laboratory manager would definitely...can be (there will always be confusion about this though) about the fact that containers are distinct ... Laboratory management software is not a requisite for a functioning lab, but it is for a scaleable lab...techs through the process. The problem? As we said above, any one of these steps can fail but, in our current...gooey contents of said containers, think carefully about how you want your software to treat those encoded...this case, the physical considerations of the laboratory make object composition a smarter choice than...

... inquiry, you always want to read before sending. 5) You can use LinkedIn before an interview, but be ...include a note about where you met IRL (in real life) and perhaps what you talked about (or how good the... this rule, but not that many). I’m asked a lot about requesting connections to complete strangers for...have been able to tell when some start-ups were about to be bought by seeing certain groups of people ...using social media as a scientists Read blog posts about networking ...

...Curr Protoc Microbiol. (2013); Chapter 14:Unit14D.5. PubMed PMID: 23686825. PubMed Central PMCID: PMC4135440...general consideration for behavioral testing in a laboratory, a high level of experimental control is required... after stereotaxic injection. Find more details about choosing AAV or lentiviral vectors in our previous...it’s crucial to appropriately time the injection about two weeks before the behavior test. This time window... Viral Vectors Resources on Addgene.org Learn about Addgene's viral service Find viral protocols and...