...Replacing the D’ region of an ITR with a sequence containing transcription factor binding sites can increase...expression at varying capacities. AAV vectors containing the AAV2 ITRs can be cross-packaged with capsids...PMID: 31914802. Savy, A., Dickx, Y., Nauwynck, L., Bonnin, D., Merten, O.-W., & Galibert, L. (2017). Impact...

... a only a single gRNA per cell. The viral DNA containing the gRNA then integrates into the genome. Thus...are poorly transduced by lentivirus, and thus screening with available CRISPR libraries was not possible...retroviral mouse CRISPR pooled library can be used for screening of cells that are refractory to lentiviral transduction...

...potential in vivo applications or multiplex CRISPR screening.   Using tRNAs Luckily, a fun feature of tRNA...given tRNA (derived from human or plant source), by tuning the length of 5’ leader sequences of a given tRNA...engineering of crops or allow for sophisticated CRISPR screenings that would perturb the unprecedented number of...

...mass spectrometry app. I digress. When I started learning proper cell culture technique, I tried to find...my lab we do a lot of genetic construction and cloning experiments. Not all of our lab enzymes are made...be beneficial for new researchers or students beginning to learn about biochemistry. The least this app...

...supernatant containing exo-AVVs from producer cells, and then to enrich the microvesicles containing AAVs (...Exosome-Associated AAV by Overexpression of the Tetraspanin CD9." Molecular Therapy-Methods & Clinical Development...

...fluorescent proteins and a new two-photon laser scanning microscope will allow Tsakanova to image cortical...insights into functional network interactions. Combining synthetic biology with engineering: glowing bacteria... bacterial sensor, he will construct plasmids containing promoter regions that activate fluorescent protein...

...media or the phones hiding under their desks than listening to me. When I got home, I saw my daughters playing...get the engagement I wanted. I therefore started planning a game that would force my students to use concepts...basic lab techniques Plasmid Land: The plasmid cloning game ...

...posts by Joachim Goedhart on the Addgene blog – outlining various important considerations when choosing...contributions (using simple web forms) while also maintaining a relatively strict database design (schema)....inaccurate data submissions, so FPbase includes a versioning system that allows users to see when a given ...

...used TRICK to examine translation of Drosophila patterning gene oskar. Early in development, the osk-TRICK...with the red reporter mRNA. Yan et al. used harringtonine, a translation inhibitor that stalls ribosomes...sequences into the coding sequence and 3’ UTR. Before beginning experiments, it’s important to verify that these...

... specific tissue type, you mate your fly line containing YFG under the control of the UAS with a fly line...often very easy to get GAL4 UAS experiments up and running with fly lines that are already available, sometimes...references and web links from my first post and start designing your first fly experiment! Many thanks to our...

...publications from these journals into two groups: those containing plasmids that the authors deposited with Addgene... other reasons for the correlation. However, determining this exactly is difficult. For example, it is... papers in the same journals! So, is all this remaining difference caused by depositing the related plasmids...

...cleave the DNA molecules at specific sites and 2) running the reaction on an agarose gel to determine the...the gel will ensure you keep smaller bands from running off; however, you may need to run the gel for longer...such as how to set up your digests and pouring/running a DNA gel.  More Plasmid Eductional Resources:...

... your academic lab, download Science Careers' "Running Your Lab" booklet. What has been the most surprising...with us every day and in the pub with us in the evening, wanting to know our latest results as soon as ...Resources for Early Career Scientists Science Careers: Running Your Lab (pdf) Advice for Choosing a Research Project...

... pair on actinin will produce different signals based on changes to the structure of actinin in low vs...associated with specific proteins of interest such as actinin. When tension is high vs. low in the cell the donor...

...forward genetics harnesses this at a large scale by screening diverse libraries of genetic variants. Several...enzyme. AID, however, only changes cytidines or guanines to the other three bases, which limits the level...bacteria. Its modular nature provides versatility in defining the mutational rate and editing window of the ...

...reached out and asked if I would be interested in joining the audit committee about five years ago. I was...of the office? Katz: I have a puppy and so I am learning how to train a dog with my daughter. It’s super... was associated with Hale & Dorr and Crowell & Morning. Karen was a founding member of Mass Medical Angels...

...across fly lines (i.e. there won’t be positioning effects). When designing experiments where you want to express...process, you will first need to do some molecular cloning. Depending on whether you are trying to overexpress...

...are commonly used for Golden Gate cloning or for CRISPR gRNA cloning. With this more functional display...display, you can start thinking about your next cloning experiment early on. Beyond these simple but powerful...

... versus eukaryote, background fluorescence). Determining practical brightness When a dish of mammalian...cells is transiently transfected with a plasmid containing a fluorescent protein (fusion) there is huge ... under realistic experimental conditions. By determining the brightness for a set of fluorescent proteins...

...Sanjana et al., 2014). It is a dual-vector system, meaning it is typically used in conjunction with a second...(Gi)-mCherry from this virus is cre-dependent, meaning that once injected, the DREADD will only be expressed... vectors and genome-wide libraries for CRISPR screening. Nat Methods. 2014 Aug;11(8):783-4. PubMed PMID...