...outlined below as well. Cas12a also has significant therapeutic promise due to the fact that it is smaller than...simplicity of Cas12a makes it easier to package for therapeutic delivery by AAV, which has strict capacity limitations...sequences of Cas12a to further optimize it for therapeutic and laboratory usage. Fast facts – PAM requirement...TTTN. Best for: homology-directed repair or therapeutic delivery.   Cas14 – Mini, but single strand ...stirred up interest for its potential as a viral therapeutic that could specifically degrade target viral ...

...maintenance - through acquisition, storage, and utilization - and on self-propagation by reproduction. Both...available or being developed to treat a multitude of therapeutic indices, including hypertension, cancer, diabetes...of functional groups of genes involved in the execution of a specific regulatory regime (such as genes...timely manner. Addgene represents an excellent solution to the problem of sharing plasmids, providing ...Nuclear receptor coregulators: Judges, juries and executioners of transcriptional regulation. Molecular Cell...

...sequence on the other strand. To guide heteroduplex resolution to favor the edit, the Liu lab turned to a strategy...inhibiting DNA repair allow for correct heteroduplex resolution? The prime editing heteroduplex is likely recognized...for optimization. They developed phage-assisted evolution strategies to evolve more effective prime editor...alternate approach to RNA stability This recent contribution by the Adamson lab also addresses the problem...mismatch repair inhibitor MLH1dn Promote desired resolution of heteroduplex     ✓ ✓ ~ ~ ~ ...Osborn, M. J., & Liu, D. R. (2023). Phage-assisted evolution and protein engineering yield compact, efficient...

...work to improve the Adv genome’s utility in research and its therapeutic potential. Note: constructs discussed...calculate the particle number: OD260 reading x dilution factor x 1.1 x 1012 particles = number of particles...assay: permissive cells are infected with serial dilutions of rAdV, fixed, and then stained 10 days later...plaque forming units (pfu), taking initial vector dilution into account. You're now ready to use your...

...dynamics, higher sensitivity to light, and better distribution of clusters without accumulation in subcellular...as starting material for the bioproduction of therapeutic methylxanthine drugs like theophylline. To facilitate...majority of this operon originates from Pseudomonas putida CBB5 and has been tailored for expression in E....under the transcriptional control of a strong constitutive promoter (BBa_J23100) and expresses N-demethylases...Holmes Jin Zhang’s lab is developing new super-resolution techniques for live-cell imaging of the dynamic...

...repeat the experiment by diluting your standard curve, performing a serial dilution of your experimental ...hydrochloric acid, which stops the reaction, turns the solution yellow. The absorbance of each well, which is ...density (OD) - the measure of light absorbed by a solution - of these wells is plotted against the known ...more quickly. Just make sure to incorporate the dilution factor in your calculation from OD to protein ...

...diseases was sponsored by Vertex Pharmaceuticals and CRISPR Therapeutics. Data shared from these trials ... Li and colleagues chose to explore CRISPR’s therapeutic potential in mice with a dominant cataract disorder...other companies have followed suite, with Beam Therapeutics initiating a base editor-based clinical trial...This ongoing trial is sponsored by Excision Biotherapeutics. Additional ongoing trials The CRISPR therapies...

...enhanced function after intravenous administration, distributing widely throughout the organism to target large.../cJ mice through LY6C1, with relatively even distribution of target gene expression (EGFP, in the depositing...transduction in New World monkeys   Figure 3: Evolutionary tree depicting the phylogenetic relationship...efficiency across species points to its potential utility in translational applications and in models such... written by Rachel Leeson, with significant contributions from Cynthia Arokiaraj. Addgene would also like...epithelium after intravenous delivery by directed evolution of underexplored sites on the AAV capsid. Molecular...Administration Results in a Differential Pattern of CNS Biodistribution in Non-human Primates Compared with Mice. Molecular...

...models for developmental processes and promising therapeutic tools. The two unique characteristics that define...cells can be used for different research and therapeutic purposes. Both types have their advantages and...of post-transplantation rejection when used therapeutically High Low (if ASCs are isolated from the patient...  While iPSCs and ESCs have similar functional utility, iPSCs bypass the transplantation compatibility...directly from a patient’s adult cells when used therapeutically, the chances of post-transplantation rejection...

...seemingly ubiquitous in nature, they are largely underutilized in research settings — mostly a consequence ...Cas3 — a tactic some CRISPR transposase systems utilize with type I-F Cascades. Figure 2: Cascade...has the potential to be incredibly useful in therapeutics. Imagine deleting a problematic copy of a gene... potential. Hopefully more people will want to utilize Cas3’s degradative power in the future! References..., J., . . . Koonin, E. V. (2015). An updated evolutionary classification of CRISPR–Cas systems. Nature...

...and Henry Siedentopf to enhance the microscopic resolution for studying colloidal gold (1).  The method ... microscope (SPIM). In order to improve axial resolution and reduce phototoxicity for imaging live specimens...using multi-view imaging can provide enhanced resolution (8,9).  Thus, LSFM is ideal for 3D reconstruction...was done by Walter Spalteholtz (10).  He used a solution of benzyl alcohol and methyl salicylate which ...later modified by others to produce Murray’s clear solution.  Murray's clear (11) was mostly used to study...

...with limited time for one-on-one mentoring. His solution? Bringing his research into the classroom. I had..., Provost partnered with colleagues at other institutions to form a support network called the MDH CUREs...MDH-focused CUREs at 50 schools, ranging from R1 institutions to community colleges, and has helped provide... and results with peers and faculty at other institutions. Provost believes these networking experiences...experiences (CUREs) as a pathway to diversify science. Evolution and Development. 24(5):127-130. Kimball, C., Powers...

... ways. Zebrafish mutants In the 1990’s, two revolutionary large-scale random mutagenesis screens produced...processes—not just development—are available from strain distribution centers in the US, Europe, and China. In addition...microscopy-based lineage tracing defined a high-resolution map of early embryogenesis in time and space,...fish lines have been developed to enable high-resolution mapping of the brain by uniquely labeling neurons.... "Pan-neuronal calcium imaging with cellular resolution in freely swimming zebrafish." Nature methods...

...are investigating how the production of cancer therapeutic drugs, vinblastine and vincristine, are regulated...transferred into the plant. In the laboratory we can utilize this natural process by replacing the genes on ...easily show which tissue have been transformed. But caution: check for GUS-like activity of your tissue. Run...or whole plants are placed in an Agrobacterium solution and a vacuum is applied with a vacuum bell or ...the dark 16 hours before the transformation. We routinely transfer our plants 16 hours before the transformation...

...commercialization office, the CIO’s office, and research computing. The two guiding principles emerging from the ...Goettingen, and several other large research institutions. The following graphic depicts the ways in which...banks and sequencing files is likely stored in institutional and lab servers specifically designed to hold...illustrated in this video, and to data stored on institutional and lab servers. Connectivity to other tools...well as the DSpace repository. These integrations utilize an interface that enables users to conveniently...

...typically carried out in one of two ways. Both methods utilize the same basic strategy: immobilize the antibody... One of the most common immobilization methods utilizes bacterial cell wall proteins, Protein A and Protein... be performed even with a pre-cleared sample.  Eluting the target protein for downstream testing Once ...washing, the target protein can be eluted. The elution method will vary depending on the specific target...with nucleic acids. Labs studying epigenetics routinely use the chromatin IP (ChIP) technique to identify...

... contaminated or misidentified cell lines. The solution Fortunately it has never been easier to conduct...working cell bank has been made by a research institution (an excellent practice), or maybe cells have ...major cell lines on public databases. Image attributions (from top to bottom) Database Center for Life...experience through working for a variety of pharmaceutical companies including GSK, AstraZeneca-MedImmune...talks and seminars on cell culture and other pharmaceutical topics. The LGC Group also provides standards...

...resultant gene targeting system could be used to routinely deliver sequences of interest to cells with efficiencies...CRISPR editing CRISPR, of course, has started a revolution in gene editing. However, while CRISPR is remarkably...the AAV approach is most well suited, since it routinely generates knockin efficiencies of 1-40% with G418...PMC4752446. 5. Asuri, Prashanth, et al. "Directed evolution of adeno-associated virus for enhanced gene delivery...nucleases with adeno-associated viral vectors for therapeutic gene editing." Precision Medicine, CRISPR, and...

...have waited. Both tasks have some utility, but I lost all the utility (fun) of eating with friends by splitting... two main things: How useful the task is (its utility) How much time it will take you to complete the...should err on the side of things that have the most utility; i.e. those things that will be most useful for...and think about what they mean in terms of the utility of your tasks. In my cell splitting anecdote, I...

... Returning to an earlier analogy, no amount of shouting will make an apple fall up. As scientist and politician...GMOs), the absence of a link between vaccines and autism, and other hypotheses mount yet policies and public...proposing to defund aspects of research at National Aeronautics and Space Agency, the Environmental Protection...Transgender issues Reproductive health Immigration Pollution Healthcare Science education To make scientists...and inclusion through funding and at their own institutions, while many also seek to honor and credit women...