...Inflammation and Immunity, as well as various combinations thereof (so called “Immuno-Oncology”) is exploding...tackle the challenge of curing cancer in cross-disciplinary projects and collaborations. Summarized by Professor...the boost it needs to help rid us of cancer in combination with chemotherapy. To dive deeper into this field...exciting development in immunotherapy is the combination of the CRISPR/Cas9 methodology with T cell receptor...

...well as an efficient mechanism of homologous recombination to facilitate simple gene replacement/mutation...directly into the host chromosome via homologous recombination. Yeast Replicating plasmids (YRp):  These vectors...some antibiotics. Auxotrophy is defined as the inability of an organism to synthesize a particular organic...selection element pairs must be utilized or a new combination needs to be created in advance of the experiment...

...convenient site for conjugation.  Finally, Fvs are the most amenable to recombinant expression and will have the...If so, you are not alone. Full-sized antibodies dominate the world of research affinity reagents, and for...not a “Fab” fragment, Fvs are often produced recombinantly and engineered to be joined by a linker peptide...

...secondary will bind to all the primary antibodies indiscriminately, giving them the same output signal. In these... Time is a precious commodity in the lab and eliminating the need for secondary antibodies would save ...use a primary antibody labelled with a probe, eliminating the need for a secondary antibody. Other immunohistochemistry... on Addgene.org Find plasmids encoding recombinant antibodies at Addgene  ...

...repair mechanisms that can be exploited to either inactivate a gene or introduce a desired genetic alteration...expression. Its catalytic domains can be mutated to inactivate the protein’s nucleolytic capability, and this...remains intact while its nucleolytic activity is eliminated (Fig. 1) (1, 3). The gRNA can be further modified...tight control over Cas9 nuclease activity while eliminating the reliance on orthogonal Cas9 species. Within...

...temperature, humidity, noise level, circadian effects (inactive versus active phase) Experimenter Interaction... of certain genes. A popular method uses Cre recombinase on transgenic flox animals to introduce modifications...- from basic types like locomotion and motor coordination to more complex situations such as learning ...Sena-Esteves M. Gene transfer to the CNS using recombinant adeno-associated virus. Curr Protoc Microbiol...

...research focuses on understanding the structural determinants of protein-DNA interactions and how these interactions...functional portion of the Nfix gene using Cre recombinase and to generate Nfix-negative neural stem cells...developmental delay, result in loss of function or dominant negative NFIX phenotypes. Developing a rapid ...

...post was originally written by Tyler Ford in March 2017 and updated by Angela Holmes and Christina Mork in..., spectinomycin, streptomycin, tetracycline, combinations of the above, or other. High or Low Copy... with highly repetitive sequences (prone to recombination in bacteria) such as lentiviral, retroviral,...

... proceeding to ultracentrifugation to avoid contaminating your transduction with the packaging cells. ...may be the result of plasmid integration via recombination during transfection, and does not necessarily...PMC84922. 5. Ugai, Hideyo, et al. "Stability of a recombinant adenoviral vector: optimization of conditions...., Michael J. Yezzi, and Barbara R. Young. "Recombination events during integration of transfected DNA...

... lab page, then onto the publication page, and finally on down to the individual plasmid page. The depositing...modifications listed can refer to changes made to the original backbone by the depositing lab, or by a previous...previously deposited with Addgene, you can list the original vector’s plasmid number here. Gene/insert: This...plasmid contains certain fluorescent proteins. Finally, for many plasmids you can find additional resources...

...(the precise mechanism isn’t well understood). Finally, TOPO cloning is another good option to consider...integration sequences, relies on site-specific recombination reactions catalyzed by BP Clonase and LR Clonase... to the use of toxic ccdB to prevent any non-recombinant vectors from propagating. The ligation-independent...since one can pool the components and define their final order through sequence homology. Similarly, Golden...

... program is the right fit for you without the financial and logistical stress that accompanies traveling...faculty, students, and each other. Before COVID-19, financial and logistical obstacles could make touring potential...COVID-19? How are students funded? Are there any financial aspects that are confusing? During a traditional...interviews for a variety of reasons (logistical, financial, etc…) now have options to fully engage in the...

... interest from its production site to its final destination. As an example, most secreted proteins are...pathway to their subcellular or extracellular destination. At the end of their life cycle, proteins may...through the vesicles to the lysosome where they are finally degraded (2, 3). Once you've created your fusion...

...expressed, they predominantly fluoresce one color but slowly mature until they predominantly fluoresce a ...of protein fusions or expressed independently) originally fluoresce green or blue and, when they mature...hallmarks of ideal FP timers: 1. Ratiometric determination of expression time The ratio of mature to immature...

...proteins of interest has occurred. Although the original Y2H systems utilized the yeast Gal4 activator,...for screening greater numbers, but the latter eliminates the step of identifying prey proteins after screening... the experiment to reduce the likelihood of indiscriminate reporter gene activation. Including a prey-... libraries can be screened using both N- and C-terminal fusions of these proteins. This way both ‘ends...

... bit of Addgene history... As you may be aware, Melina Fan, her brother Ken Fan, and her husband Benjie... corn toss boards, Kan Jam, and Ladder Ball (I finally discovered my hidden talent in Ladder Ball... there...popularly- cookies). 3. Do something outside of your ordinary environment Team building events like Escape the...Veckerelli is the Administrative and Operations Coordinator at Addgene. In addition to her operational responsibilities...

... nuclease domain. These mutations result in an inactive RNA-guided DNA binding protein (dCas9). dCas9 ...the next 3 ranked sgRNAs. Dolcetto was able to discriminate between non-essential and essential genes similarly... an improved activator design that consists of inactive Cas9-VP64, a sgRNA with two MS2 RNA aptamers, ... active sgRNAs for CRISPR-Cas9–mediated gene inactivation." Nature biotechnology 32.12 (2014): 1262. PubMed...

...last month it’s been viewed by users in the US, China, India, Canada, the UK, South Korea, and beyond!...protocols So that’s great, but what if someone in China improved upon that protocol? How would the thousands...prior to publication, and this will delay the dissemination of your improved protocol. Many scientists, ...need for quicker, more effective updating and dissemination of experimental protocols, simultaneously allowing...

...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...

... in directly controlling muscle activity and coordinating movement, several additional brain areas that...dark areas (protected) as opposed to brightly illuminated ones (risky). They also prefer to explore novel... (Howerton at al. 2014). Addgene has several recombinase-expressing ready-made AAV vectors which can be...scientists study such as sleep, food intake, mobility/coordination, learning, and memory. We’ll cover these in ...