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... Originally published Jul 14, 2015 and last updated Sep 16, 2020 by Beth Kenkel.  CRISPR genome editing...catalytically dead dCas13b to the catalytic domain of RNA deaminase ADAR2. Constructs containing the ADAR2 truncation...orthologs works for some applications, it doesn’t eliminate the need to deliver larger cargo such as base ...Cytosine and adenine base editing of the brain, liver, retina, heart and skeletal muscle of mice via adeno-associated...

...but some applications have been challenging or inaccessible. The Ting lab recently made two deposits that... Figure 3: A) LOV-Turbo becomes active when illuminated with blue light, catalyzing the biotinylation...that had been trafficked from the “source” to “destination”. Images adapted from Lee and Cheah et al. 2023.... (2019). A toolbox of IgG subclass-switched recombinant monoclonal antibodies for enhanced multiplex ...

...Microbe meeting, but it was my favorite because I finally felt like I had a network of people to talk to ..., Kat Milligan-Myhre, and Pragati Chengappa.  Financial and moral supporters Denise Akob, Sara Alexander...Heather Hendrickson, Jonathan Jacobs, Mo Kaze, Christina Kellogg, Heather Martinez, Pat Schloss, Tara Smith...McGovern, and Julie Wolf. Apologies to anyone I’ve inadvertently left off this list. Many thanks to our guest...

..., homology dependent repair (HR, homologous recombination; SSA, single strand annealing) took place and... with RNAs encoding Cas9 into the oocyte. This original method was developed as the use of CRISPR/Cas9... RNA and therefore there has less chance of contamination with RNase leading to degradation that can set...pups. Taking a toe, a finger, a tail tip, or a combination of these will allow you to determine which pup...

...circuit, while a nose-poke at a separate hole (inactive hole) does not. If activation of the circuit is...unwanted phenomenon or it could reveal something fascinating. Behavior is based primarily on careful observation...experimenting and hopefully optogenetics will help illuminate (is that pun worn out yet?) your research. Derek...PMCID: PMC4165888. 6.Witten, Ilana B., et al. "Recombinase-driver rat lines: tools, techniques, and optogenetic...

... practice, the term "promoter" describes the combination of the promoter (RNA polymerase binding site)...synthetic promoters, which typically include some combination of other promoter elements, and tend to be more...expression mRNA Mammalian promoter from phosphoglycerate kinase gene.  Constitutive  Widespread expression, but...optogenetics mRNA  Ca2+/calmodulin-dependent protein kinase II promoter Specific Used for neuronal/CNS expression...

...This post was originally written by Melina Fan and updated Nov 3, 2022 by Susanna Stroik. You’ve designed...outlined below, the technique can be used as a preliminary validation on your bulk population. This way ...TIDER (Tracking of insertions, deletions, and recombination events). TIDER can screen for specific homology-directed...

...To acquire these measurements, the cytometer illuminates the passing cell with a laser and detects the...colleagues.   Figure 1: The laser pulse illuminates the cell and is scattered forward at a low angle...represents the time and intensity of the cell’s illumination (see Figure 2). H describes the maximum signal...For our last example plot, I will show you a combination of two stains, in this case through conjugated...

...the correct size, and purify the DNA fragments. Finally, you simply incubate these three fragments together...loop (as might be anticipated to occur within a terminator sequence), since this would directly compete ...one step with no scar sequences remaining in the final product. The long overlapping region between fragments...different fragments. One way around this is to use a combination of standard sequences with overlap regions, such...

...engineered proteins responsive to light in combination with retinal) variations designed to give researchers... R., Gunaydin, L. A., Hyun, M., Fenno, L. E., Gradinaru, V., Yizhar, O., & Deisseroth, K. (2012). Principles...

...then be exposed to a lysis buffer to release recombinant proteins directly onto the nitrocellulose for...receptor binding protein hit and can go back to the original plate to identify which phage gene encodes that... 1 M NaCl in order to enhance folding of the recombinant proteins. Optimal salt concentration will vary...colony expressing a functional binding protein was originally located. Now you can grow up your colonies of...

...American system), in 2009 (at the beginning of the financial recession) I knew obtaining work in scientific...facility, where I tested materials for bacterial contamination. It was a really interesting job at first, everything... are rapidly expanding, as this might not be sustainable. After working at this company for several years...would produce the best outcome for a specific combination of genetic mutations. This is a particular challenge...

...testing several vectors and packaging cell line combinations, Miller et al. showed that infectious wild-type...are capable of transducing certain quiescent or terminally differentiated cells such as macrophages and ...research due to the possibility that mutation or recombination with human retroviruses could lead to RCL. ...of retrovirus packaging cell lines to avoid recombination leading to helper virus production." Molecular...

...includes: Michelle, the Senior Scientist who coordinates all kit creation and quality control efforts ...once famously said “I think this is the most extraordinary collection of talent, of human knowledge, that...successfully doled out to multiple people: someone to coordinate, someone to decide, someone to delegate, etc....Next Great Discovery is exactly that: a myth. The original Dr. Frankenstein from Mary Shelley’s novel worked...

... contest is now closed. Stay tuned to see the finalists and vote for your favorite! Our mascot who has...neutral name. The Addgenies will choose up to four finalists and we’ll ask you to vote for your favorite name...We hope you submit your best ideas because the finalists will receive some newly designed Addgene swag....

...Addgene. Boston, MA, USA 42.3601°N 71.0589°W – Alina Chan The job of scientists to observe how things... and generation is in the veracity of its work. Alina Chan is a postdoctoral researcher at Harvard Medical...saying that climate change is a hoax. This has culminated in the current President holding and campaigning...interactions, and science education. Photo cred Alina Chan. Seattle, Washington, USA 47.6062°N 122.3321...

...structures are more stable and local movements predominate (e.g. a basal progenitor cell migrating to the...fluorescent proteins (Salomonnson et al. 2012). Finally, it is worthwhile to test the photostability of...used to study B and T cell dynamics within the germinal center of murine lymph nodes. However, one must...marking and tracking cells is the the Cre/lox recombination system. A researcher can flank fluorescent proteins...

...and important details that you won’t find in the final publication and we have already collected over 100...accelerate your research. Sincerely, Melina & The Addgene Team ... Melina Fan...

...renewable electricity is certainly not the only sustainability effort we’ve implemented at Addgene. The kitchen...check out Leila Haery’s blog series, “Bringing Sustainable Practices to the Lab.”  Whether participating...blog Subscribe to the Addgene blog Read our sustainability in the lab blog posts Read another blog post...

...Working on the Eye 9:05-10:16 : Cell Targeting in Retinal Gene Therapies 10:17-14:34 : Progress in Treating...type-specific manipulation with GFP-dependent Cre recombinase.” Nat. Neurosci. 18(9) (2015): 1334-1341. PubMed... retina...