...experimental approach you choose, it’s essential to consider the technical limitations of each approach and to...affect the phenotype. Viral vectors and genetic approaches are continuously refined and developed so there... Delivery of AAV into the brain can usually be achieved by a directed brain injection, but some serotypes.../loxP or Flp/FRT, with AAV vectors are common approaches to control transgene expression by restricting...types. Specific transgene expression can be also achieved in nontransgenic animals by using cis regulatory...post-translational regulatory elements. Examples of these approaches are given in Tables 3-7. 3. Making use of AAV...populations using viral strategies Combining different approaches can increase the specificity of cell targeting...

... executive director of Central Indiana Science Outreach and a Postdoc at Indiana University School of ...Biology, I realized I wanted a career in science outreach: engaging the public to promote an awareness and...founding a nonprofit called Central Indiana Science Outreach, or CINSO. We organize fun science events for ...do fun science activities with the public, so I reached out to people who were educating the public and...and learned a bit about recruiting scientists to outreach.  Try new things to further your career Take advantage...new people. There were visiting seminar speakers each week, my department and graduate office arranged...Especially as someone interested in doing science outreach, not only can I easily share science with the ...

...resulting in processing by the endogenous RNAi machinery. Alternatively, endogenous miRNA precursors can... either technology was equivalently precise, but each method recovered different genes known to be required... two. Consequently, I will focus on different approaches and considerations present at the first stage...initial state of the dsRNA upon activating the RNAi machinery (long dsRNA, typically hairpin RNA, or artificial...long-term RNAi phenotype. Long hairpin RNA The easiest approach to induce RNAi involves the expression of long...different constructs are available to the community. Each construct has unique features, such as inducible...RNA The greatest limitations of the long hpRNA approach are thought to be off-target effects and variable...

... passes through the machine’s lasers and gives off a signal read by the machine’s sensors. (Usually this...could be debris or unbound antibodies.) Each event read by the machine is plotted as a dot on an X,Y axis ...plotted on an invisible Cartesian plane (right.) Each event is plotted by the amount of signal read from...variations in fluorescent signals, and how the machine reads/interprets the signals, the axis is instead...populations. This shape is called a ‘gate’; it tells the machine that everything inside the gate is one population...doublets and single cell populations are very close to each other.        Figure 4: Gating strategy ... numbers representing the percentage of cells in each gate. Used under Creative Commons license.  ...

...single model. The first approach can lead to new directions, while the second approach could terminate with...the problem and goal, the approach, anticipated outcome, and payoff for each aim. Learn to communicate...preparing for both types. Your research vision is your approach to resolving a significant knowledge gap in your... you need to be able to describe experimental approaches in terms of the question, the methods, the tools...question, the anticipated outcomes, the alternative approaches, and what will you do if your model is correct..., a clear easy-to-understand question/goal, an approach, anticipated outcomes, and preferably a description...successful. Note that aims do not need to be grant aims. Each aim could be the basis of a single grant. Either...

...Celano of Propel Careers and Rachel Casseus, Esq. Founder of Casseus Law. Rachel Casseus Esq., Founder Casseus...guest bloggers Lauren Celano of Propel Careers and Rachel Casseus, Esq. Founder of Casseus Law. Disclaimer...numerical quota of 85,000 total H-1B visas approved each fiscal year: 65,000 visas plus 20,000 reserved for...and the arts, and requiring the attainment of a bachelor's degree or its equivalent as a minimum (8 U.S....). The foreign worker must possess at least a bachelor's degree or its equivalent and state licensure,...should only focus on your current scientific achievements. Participating in the scientific review process...international recognition for your outstanding achievements in your specific academic field, (2) that you...

...community, with approximately 100 new CRISPR depositors each year.    Figure 1: Addgene CRISPR distribution...CRISPR tools people were depositing and requesting each year (Figure 2). We did a similar analysis with ...function, by year. (B) Number of requests per month for each CRISPR function from 2012 to 2023; data are smoothed...libraries deposited and requested grows steadily each year. (The dip in 2020 is likely due to the pandemic... style When we plotted CRISPR plasmids requested each year by year of deposit (Figure 3), we saw that ... plasmids (by age) are requested and shipped out each year.     Figure 3: Percentage of requests...plasmids requested, categorized by year of deposit, for each year from 2012 to 2023.  What does this tell...

...be needed to achieve your intended next career stage and create a timeline for achieving these goals. ...and teach classes. Therefore, seek out management and conflict resolution workshops. Get teaching experience...the position, you may also want some evidence of teaching experience. How do you get there and more practically...you constructive feedback on your research, and teaches you how to write as well as mentor. Yet, even the...not always available for mentees due to travel, teaching, grant writing, etc. Also, it is unlikely that...perspectives, a shared communication style. You “get” each other. The other factor is how available the potential...writings. 8. Learn to mentor, manage, communicate, and teach The dirty secret about becoming a group leader is...

...microscopy (FLIM) method. Acceptor photobleaching method The acceptor photobleaching method is simple but limited...An EAMNET Teaching Module http://www.embl.de/eamnet/downloads/modules/FRET_teaching_module.pdf Day RN, ... knowing if these students know or interact with each other. FRET reduced the scale of our perception ...spectrum overlap. If you choose a pair too close to each other in the spectrum, you can easily directly excite...contrary, if you choose a pair that are too far from each other, you don’t have any cross-talk but you also...experiment. You can then choose the most efficient approach for your own system. The simplest and the most...fluorophores does exist in the real world and corrective approaches and appropriate controls are required to make...

...endogenous molecular target, meaning the dye will be attached to the target through binding of the molecular...encoded tag is required. These dyes can also be attached to an antibody to detect various targets, such...or glycans. Fluorophores can also be covalently attached to self-labeling proteins. Some common examples...covalently reacts with a small substrate ligand motif attached to a fluorophore. When paired with a protein of...higher specificity, increased versatility in probe attachment, and compatibility with far-red and near-infrared... must be taken to assess if proper staining is achieved. Fluorescent ligands have more flexibility due...peak wavelength can also decrease the rate of photobleaching. Interaction with receptors: Understanding ...

..., but you’ll only spend half of each day on it. The other half of each day you can spend accomplishing...how much time I have blocked off for each project and how useful each item is. For instance, if I don’t ....e. those things that will be most useful for achieving your goals. However, you should have a good mixture...experiment, but you’ll have to spend the entirety of each day doing the experiment. The second way, it will...fails, you’ll still have made small accomplishments each day that should help keep you content and give you...thinking about when or how long it would take to achieve these goals. I have far too many projects to operate...Knowing this, I hand write to-do lists at the start of each day to remind myself of the specific activities ...

...gRNAs on the same plasmid, you’ll make sure that each cell that gets the plasmid contains all of the desired...for processing a continuous multi-gRNA transcript, each gRNA must be expressed from its own promoter. But...multiplexing is to clone the oligonucleotides specifying each gRNA target sequence into distinct expression vectors... using the enzyme BbsI. These expression vectors each contain Type IIS restriction sites flanking the ...but they’re optimized for different purposes.   Gersbach Lab multiplexing plasmids: This plasmid set allows...generate four unique kanamycin-resistant plasmids, each containing a different gRNA target sequence downstream...you’ll clone in a polyT-termination sequence for each unused promoter. This step is necessary to generate...

... final wake it all settles down and the value of each item changes from what the reviewers so painstakingly...subjectively ascribed relative to the basic needs of each individual; the  community as a whole has yet to...and it means that there is no one-size-fits-all approach. You will have to select a tool based on your ...opinion. I could spend a lot of time running through each tool in detail, but that is already covered in the...the tools into the individual features that make each tool unique. The CRISPR Software Matchmaker is composed...organisms: This section lists the genomes supported by each tool. The genomes are denoted by organism name and...CRISPR experiments evaluate the predictive power of each tool they are interested in by comparing their past...

...transcription machinery.   Well, I’ve got some good news: complete DNA demethylation can be achieved by dCas9...can be achieved by fluorescence-activated cell sorting, and stable gRNA expression can be achieved using...simpler, yet more effective, CRISPR-based epigenetic approach (spoiler alert: it’s dCas9… with no modifications...). But there are a number of drawbacks to this approach: TET proteins are not demethylases; they are ... activity. They interact with transcriptional machinery to modify local gene expression, confounding any...and Szyf 2021).    Figure 3: A simpler approach - dCas9 blocks DNMT1 methylation, leaving the ...eLife.34870.  Kaas GA, Zhong C, Eason DE, Ross DL, Vachhani RV, Ming G-L, King JR, Song H, Sweatt JD (2013...

... At the end of the run, each droplet is read as positive or negative for each reaction, math happens (...and departments will have qPCR machines available for use. ddPCR machines are not as commonly available...molecules added to the mastermix that are read after each cycle. The raw data is then normalized to a reference...droplet immersion technique, courtesy of CAR15.) Each droplet contains a primer and probe set for both...can be confidently interpreted as integers. Since each sample is partitioned into thousands of reactions...experiments, a number of factors will determine which approach is right for your experiment. You’ll have to consider.... PMID: 21554069. Haridan, U. S., Mokhtar, U., Machado, L. R., Abdul Aziz, A. T., Shueb, R. H., Zaid, ...

...’s lab at the University of Tokyo and Charles Gersbach’s lab at Duke University developed similar systems...light-activated CRISPR/Cas9 effector (LACE) system, the Gersbach lab utilized a similar strategy to develop an ...fusing a photoinducible, heterodimerizing domain to each of the Cas9 fragments, the authors created a photoactive...and scattered by the skin, so it is not able to reach deeper tissues. Additionally, high intensity exposure...photoswitches Photoactivation of Cas9 can also be achieved using single-chain fusion proteins by fusing Cas9...saCas9) proteins, showing the versatility of this approach. This study generated the first optically active...control over Cas9 activity. Dominik Niopek’s lab achieved this by fusing the anti-CRISPR Acr protein, AcrIIA4...

...going away parties and potlucks for each other. The Church Lab enjoys each other’s company outside of the ...productive and successful research lab. Once goals are reached, how do science labs celebrate accomplishments ...work-life balance. How do you celebrate your lab’s achievements? Tom Ellis highlights the importance of acknowledging...to eat out together. “We also have a lab day trip each year and as we do a lot of yeast research, a few...Although hard work and perseverance are required to achieve success in the lab, it is important to take the...along the way. Take time to celebrate the lab’s achievements and enjoy time with friends and family. And ...fun outside the lab! How does your lab celebrate each other's accomplishments? Any tips on making time...

...serviceman in the U.S. Army, and hockey coach all shaped his teaching style, with the former offering excellent...interpreted this trend as evidence that the best teachers and mentors pass on more than just technical knowledge...University of San Diego), decided to take a different approach. His own most impactful learning experiences happened...opportunities for his students. But he also found that his teaching responsibilities left him with limited time for...while reminding participants of the value of this approach and encouraging them to engage in the problem-...encourages faculty mentors.)  Collaboration Maximizes Reach Many new professors are wary of the time and effort...in setting up a CURE, especially if it’s a new approach within their department. So in 2016, Provost partnered...

...same cloning procedure can be used to incorporate each element into the final construct. This strategy ...Four individual expression constructs were created, each containing a unique fluorescent protein with minimal...minimal spectral overlap between them (Table 1). In each construct, 3 copies of the fluorescent protein were...enabled labeling of particular structures (Table 1). Each construct was flanked with a CMV promoter (to drive...labeling, with strong expression and detection of each fluorophore at the respective subcellular structure... 2) are contained in individual plasmids and are each flanked by the same multiple cloning site (MCS)....pattern of the MCS is regenerated at the flanks of each ligation product, which is the basis for the repeatability...

...glimpses into your personality that a list of achievements can't”. John Little, Managing Director of Successful...listing all your professional experience and achievements), but also to tailor and make your cover letter...particular skills required, and the expectations for teaching and other university obligations. If you really...postdoctoral position so I thought it might be good to reach out to you.   Dear [recruiter name], I would like...closely as possible. Always support skills or achievements with measurable facts and do not forget to explicitly... future research plan and new ideas, plus your teaching experience. In the case of non academic positions...formulas. Tailor a totally different cover letter for each position. Everything you want to say should fit ...