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...gamma-retroviral packaging plasmids are not interchangeable. General envelope plasmids, such as VSV-G, ...that can reveal how genetic alterations lead to changes in phenotype. The plasmid comes with a 1.9 kb stuffer...is a 3rd generation lentiviral backbone from the Zhang lab expressing S. pyogenes Cas9 and a gRNA cassette... original lentiCRISPR v1. Special note from the Zhang lab: We are constantly improving our CRISPR reagents...

...Shoemaker, D.D., Astromoff, A., Liang, H., Anderson, K., Andre, B., Bnangham, R., Benito, R., Boeke, J.D....to a whole genome duplication and extensive rearrangements in the S. cerevisiae lineage (7). In our class... the Advancement of Science. (2011). Vision and Change in Undergraduate Biology Education: A Call to Action...

...References and Resources References Chen X, Choo H, Huang X-P, Yang X, Stone O, Roth BL, Jin J (2015) The First... endogenous gene expression, which can lead to changes in normal cell physiology and function (Saloman...Analgesia, as well as Ligand-Independent Functional Changes in Sensory Neurons. The Journal of Neuroscience...

...edited and the sequence of the edit, so its design changes for every new prime editing application. And optimizing...although sequences with G/C content outside this range can still be optimized (Anzalone et al., 2019). ... Homology between the pegRNA scaffold sequence (orange) and the genomic target can increase the odds of..., S., Shehata, S. I., Dohmae, N., Ishitani, R., Zhang, F., & Nureki, O. (2014). Crystal Structure of Cas9...

...., Németh, J., Nagy, Z.Z., Wu, K., Wu, R., Xiang, L., Fang, X., Jin, Z., Goldblum, D., Hasler, P.W., Scholl...as inhibitory neurons, rods, cones, or retinal ganglion. Rather than using naturally occurring promoters...Cre-dependent-GFP expresses in horizontal cells and ganglion cells, only the horizontal cells transduced by...

...insertion, inversion, translocation or cassette exchange. By utilizing recombinases to manipulate genomes...Optogenetic tools use light to induce conformational changes in a protein.    Cells expressing an RFP...delivery constraints and scientists seeking a wider range of targetable genomic sequences could benefit from...10.1126/sciadv.abb7438 The CRISPR corner By Jennifer Tsang Here are a few highlights from recent CRISPR plasmids...

...Indicate the amino acid change as well as any functional consequences of that change. For example, “Mutated...Hazardous If the plasmid will produce anything dangerous or toxic to humans in bacteria, select “yes” in...written by Tyler Ford in March 2017 and updated by Angela Holmes and Christina Mork in February 2024.   ...

...Forum 2015 from Joanne Kamens, PhD Thoughts on Guangzhou and the Science City The conference was located... new “Science City” area on the outskirts of Guangzhouon west of beautiful Mao Fengshan Forest Park. As...locate there. I made time to go into the city of Guangzhou and see some of the sites. Most fascinating to... CRISPR/cas9 and more are being reported. Zhou Songyang from Baylor College of Medicine and Sun Yat-sen...

...standards for hybrid, virtual, and in-person events change seemingly daily (and are inconsistent between universities...both sets of interviews. I also had universities change their format of interview from a travel day to ...response to challenges on their end. Though the quick changes in plans was stressful in and of itself, the flexibility...everyone involved in the process. COVID-19 has changed many aspects of academia, and the PhD application...

...you to better analyze your results; it does NOT change anything in your samples. This means that, unlike...Figure 1B). For single-color samples, you can only change the compensation values for whichever fluorophore...can click on the correct box and either manually change the compensation values or adjust the values with... help you get started. Remember, you can always change your compensation values during the analysis portion...

...pipetting error or an incredible discovery. The range of RQ values used to call copy numbers should be...confident determination of copy number, albeit within a range of RQ values as noted above.   In both applications.... Muhamad P, Chaijaroenkul W, Congpuong K, Na-Bangchang K. SYBR Green I and TaqMan quantitative real-time...

...adenosine overhangs left by Taq perfectly anneal with the topoisomerase I generated overhangs in the backbone...sites allowing you to easily create customized overhangs to assemble up to 6 different fragments in a prescribed... coli then fuse together the PCR products via overhangs left over from the PCR process (the precise mechanism...

...in that sense, models can be both useful, and dangerous. Subscribe to CRISPR Blog Posts How do educators...slow down and focus their attention on something tangible. Under the guidance of a trained educator, these... we jump into a new modeling project can be a dangerous thing. Fortunately, Jacob was able to quickly ... This backbone model of Cas9 is based on Fuguo Jiang’s structure of Cas9 – primed for cleavage. This structure...

... Protocol Exchange Website: http://www.nature.com/protocolexchange/ Nature Protocol Exchange (NPE) is ...your favorite protocols for future reference. The exchange currently boasts over 2,500 protocols covering...

...increasing diversity and inclusion in STEM disciplines (Bangera & Brownell, 2014; Duboue et al., 2022), as they... now connects MDH-focused CUREs at 50 schools, ranging from R1 institutions to community colleges, and...instructors, and provides opportunities for students to exchange ideas and results with peers and faculty at other...classroom.   References and Resources References Bangera, G., Brownell, S. 2014. Course-based undergraduate...

...reduce off-target effects of gene therapy, Feng Zhang’s lab has developed a system called DIRECTED (Delivery...Design). This modular approach combines an interchangeable set of fusogenic and targeting components to...where the fusion and targeting components are interchangeable. Image from Strebinger et al., 2023, reused...be tethered on the viral surface, expanding the range of targetable cell types. DIRECTED is compatible...

...skipped or whether selecting colonies and performing Sanger sequencing will suffice…and the answer is no. Due...screening - An excellent protocol paper by the Feng Zhang lab describing construction, lentiviral production...CRISPR/Cas Systems Forum- Moderated by the Feng Zhang lab, this is a useful place to ask experimental ...Abudayyeh OO, Platt RJ, Brigham MD, Sanjana NE, Zhang F. Genome-scale CRISPR-Cas9 knockout and transcriptional...

... often weren’t easily accessible, due to a wide range of logistical barriers. Addgene worked to identify...availability vs accessibility issue was found in a less tangible form: knowledge and information about the materials...is achieved through interconnected incremental changes. It was only after creating a plasmid repository...learned about the importance of smaller, systemic changes, like using alt-text, captions, and inclusive design...

...General Meeting seven other times I had subsequently changed fields and my cohort of friends and colleagues ...in Atlanta, GA, in 2018. A lot of my experiences rang true with theirs.  It was at this dinner and the...Singletons logo. Image: James et al., 2020. Help me stranger - Finding mentors for the meeting Just like analysis... cells regulate gene expression in response to changes in cell density. At the mixer, they meet other ...

...experience will be a little different. Some of these changes you may enjoy immediately and some of them may ... just take some getting used to. Just remember: change is good. You can experience a real onboarding...re also getting the opportunity to learn their “language”, and in doing so you may discover that your career...eventually. If you approach that transition dreading the change, the adjustment period will be that much longer...