... effect different perturbations. Consider, for instance, the challenge of attempting to simultaneously...without making any functional changes to Cas9. For instance, when native (nuclease-competent) Cas9 interacts...directed to a target upstream of the transcriptional start site (TSS) of a gene of interest, it induces potent...directed to a site downstream of the transcriptional start site of a gene of interest, Cas9 forms a steric ...

...plants that inherited an edited gene to confer resistance to the herbicide chlorsulfuron (Homo line 2) ... molecules, which are much brighter and more photostable than fluorescent proteins, making them ideal ...dendrite. D) Fluorescence signal as a function of the distance between measured pixel and the soma. The initiation...postmortem brain samples. Results show positive staining in tissues from patients who had confirmed neurodegenerative...

...that might serve as a starting point for developing useful tools. A lucky start As Andrew and Maria explained...questions alone. They readily admit they don’t fully understand the physical mechanism, though they have collected...luck) to get here, he believes MagLOV is just a starting point for much more. If you’re interested in working...

...crucial background. Practice a few times while standing at your poster – it’s a different experience to...your computer screen than it is to pretend you’re standing at the session. Present with confidence Now that... asked a question that we don’t fully hear or understand what we are being asked. I am personally guilty...one. Take a deep breath, and if you don’t fully understand the question, ask for clarification. Having them...

...days of structural biology, Byron Rubin, a crystallographer working with Jane Richardson, provided an ... model of the protein in your hand – you will understand the power of a physical model. Until then, you...turn it over and over in their hands. They then start talking to themselves as they compare the features...Under the guidance of a trained educator, these instances of focused attention become magical teaching moments...

...a physiological effect of SARS-CoV-2 toward destabilizing one’s natural immune response to Adenovirus ...Adenoviral vectors are commonly derived from the mastadenovirus members (mammalian-infecting members) of the...delivery and for oncolytic virus in addition to standard uses in gene therapy and in basic research, like...death protein (E3-11.6K) is required at very late stages of infection for efficient cell lysis and release...

...curriculum. It’s through the PTA that I recently started coordinating the annual Math & Science Night, a...scientist. I gave a timeline of my science life, starting back to Mr. Kelley, my 7th grade biology teacher...them had imagined for a scientist. I think that establishing this history helped demonstrate that a) I was...curriculum and offered engaging examples of how understanding genetics is so timely.” Genome engineering, ...

...Our story today starts with a bit of Addgene history... As you may be aware, Melina Fan, her brother ...buidling - they relieved some of the pressure of start up life and maintained their sanity by throwing ... at Executive Director Joanne Kamen’s home- a testament to the familial aspect of Addgene. As our size...will avoid chaos and delays in getting the fun started. I’m aware that  “organizing the fun” sounds a ...

...protocol. Many scientists, publishers, and small start-ups are turning to software apps to solve our need...are just a few promising options to get your lab started:   7 ways to share your lab protocols online Addgene...Check out all the benefits and how you can get started on our deposit page. Protocols.io Website: http...satisfaction of checking off each step as you painstakingly complete your experiment – one small victory...

...similar or better efficiency than PEmax, the current state-of-the-art. PE6b was very efficient for short, unstructured...fluorescently labeled. Using lentiviral vectors to stably or inducibly express tagged iPAK4 from different...distribution and has been verified to perform well in standard IHC and western blot protocols. You can find more... Figure 5: Delipidated mouse brain tissue was stained with Anti-PSD-95 [K28/43R] (70 µm mounted sagittal...

... the lab somewhat mirrors the complexity of understanding their wild counterparts. Those familiar with...process that isn’t as simple as it seems. Take, for instance, the GCaMP-encoding viral vectors available in...revamped our viral vector selection process to standardize our feedback and evaluation of viral vectors....requestors can upload their experimental data in a standardized format, sharing how well a given viral vector...

...original formatting. In this way data is moved from a static and isolated vehicle and becomes part of the dynamic...permanent research record, yet they are crucial to understanding research and the thinking and workflows that...third-party access to the ELN arises here. Notwithstanding access restrictions based on permissions, however...it would be possible to grant those wanting to understand and/or attempt to reproduce research relating...

...purpuratus, the spiny purple sea urchin. It’s not a standard model organism, but it was easy to harvest eggs...this wonder of the organism.   It’s not an overstatement to say that CRISPR/Cas9 editing is one of the...which these methods are presently unavailable.” As stated by one of the NSF program directors, this program...Now what? What would you study first? They are starting by seeing if they can change long legged lizards...

... Cas9 orthologs have been isolated, and our understanding of their PAMs is shown in the table below.  ... Streptococcus pyogenes (Sp) 3' NGG Staphylococcus aureus (Sa) NGRRT or NGRRN Neisseria ... useful than SpCas9. For example, Cas9 from Staphylococcus aureus (SaCas9) is about 1 kilobase smaller...Zhang F (2015) In vivo genome editing using Staphylococcus aureus Cas9. Nature 520:186–191 . https://doi.org...

... For every experiment, an investigator needs a standard against which the results can be compared; results...inconclusive and unreliable. Proper controls provide the constant variables that enable the correct interpretation...should be included as these provide the baseline/standard against which other samples can be compared. The...least you should repeat your whole experiment from start to finish multiple times and on different days. ...

... paper, Sekar and Periasamy defined FRET as "a distance-dependent physical process by which energy is ...fluorophores used are in close vicinity. Thus, the distance between the two fluorophores and their orientation...less important when designing a biosensor, as the distance between the two fluorophores and their spatial...measurement methods as feasible when first beginning to establish the FRET methodology for a given experiment. You...

...Stroik January 24, 2023. DNA damage drives genome instability and contributes to cancer, premature aging, and...efficiency for more details on how "Cut-to-Mutation distance" affects editing efficiency. One important point...moderated by the Zhang lab CRISPR Challenges: Standardization and HDR CrispyCrunch Optimizing Donor DNA for...DNA double-strand break repair pathway choice. Shrivastav M, et al Cell Research (2008). PubMed. Mechanisms...

...cancer-causing mutations into the endogenous allele of the STAG2 tumor suppressor in human cells (Kim et al, 2016...compared to CRISPR editing CRISPR, of course, has started a revolution in gene editing. However, while CRISPR...applications that require large gene fusions. The current state-of-the-art in combining the AAV and CRISPR-based...human cells harboring tumor-derived mutations in STAG2." PLoS genetics 12.2 (2016): e1005865. PubMed PMID...

... a mutation introduced by CRISPR. The sequence started clean and then, about 100 bps into the run, it ...CRISPR for mouse genome engineering From the very start, generating mouse models using CRISPR is easier ... genome. However, with only 3.4% of founders establishing germline transmission of the transgene, this...Efficient Production of Gene-Modified Mice using Staphylococcus aureus Cas9. Sci Rep 6: . https://doi.org/10.1038...

...across the genome) has been implicated in disease states, and targeting the epigenome may make certain processes...has fused LSD1 to the smaller nuclease-dead Staphylococcus aureus Cas9 (Sa-Cas) and deposited plasmid ...more than dCas9-VP64, especially when targeting distal enhancers. As the effects of these tools are likely...T, Rivenbark AG, Rashwan R, Blancafort P (2015) Stable oncogenic silencing in vivo by programmable and...