...succeed in communicating. Even the most rigorous, insightful, and novel scientific research will be wasted...your past work has prepared you to contribute at a high level to their team.  When you're writing a grant...anyone else in the world to do it.  In the lab, it might mean persuasively articulating why the results you've...to be reviewed by interdisciplinary panels.  And highly successful scientists need to communicate with ...

...BioBuilder Educational Foundation in partnership with high school teachers to boost interest and understanding...what started in Cambridge with a handful of local high school teachers receiving training modules in synthetic...fall:  The BioBuilderClub BioBuilderClub engages high school teams around the world who combine engineering...and STEM-loving students include The BioBuilder High School Apprenticeship, preparing local (Massachusetts...

...instrumental in helping us establish consistent, high-quality protocols for lentiviral production. Bryan...Janelia Research Campus have provided valuable insight into AAV production. Finally, we are blessed to...surveyed hundreds of scientists about various ways we might expand Addgene’s services. There was an overwhelmingly...helps scientists and allows us to maintain Addgene’s high quality standards?” Under the leadership of Addgene...

...disease. CRISPR has made genomic SDM relatively straightforward, but plasmid-based SDM has lagged behind. While...well, but poor template digestion can result in a high number of background colonies carrying the unmodified...plasmid is not maintained. Recombination efficiency is high enough to negate the need for selection usually ...introduce mutations. Using commercially available high-fidelity polymerases such as KOD and Phusion, which...

...labeling tubes never takes longer than 5 minutes, right?) As a result, I’d always end up stressed and rushing...add in personalized steps like “check drawers for light-sensitive reagents” or “re-sterilize now.” Have ...Moving your reagents from your left side to your right as you use them can help you remember what's been... Fig. 2: Blugene moves reagents from left to right as they use them.    Prevent Interruptions If...

...Viral integration of a construct is also reliably higher than traditional transfection and random genomic...transfection method. This is because viral systems have a high infection frequency and the viral titer (the number...responses in addition to antibody response 2) very high immunogenicity 3) and long-lasting immune response...treatment and gene therapy have also been in the limelight. The FDA approved the first oncolytic viral therapy...

...Mendenhall: The benefits are two-fold: CRISPR/Cas9 offers high efficiency of inducing double strand breaks and ...weeks. CRISPR saves you a couple of months. It’s higher throughput and faster. It’s also easy to run things...transcription factors, but there are many reasons you might want a system like this. It’s actually surprising...with this antibody problem. We thought it was the right thing to do to put these out there. It’s a simple...

...interested in following? Someone you admire for their insight and way of expressing it in meetings? The more ...advice. Your family and friends know you well and might have good ideas for who you should connect with....cautious and make sure to choose one that comes highly recommended from someone you trust. A good coach...compliment to ask someone to share their wisdom and they might just not have the time. Thank them anyway and move...

...concentrations. Higher IPTG concentration leads to higher RepA expression which results in high plasmid copy...repressor because TALEs can be programmed to bind tightly to arbitrary DNA sequences and have been shown ...

... is weaker than reporter staining. Picking the right reporter for your RANBody While all three reporters... fluorescently labeled tyramide molecule into a highly reactive radical, which is then covalently linked...antigens. Spaghetti Monster Spaghetti Monsters are highly antigenic tags that have 10 HA or 10 MYC tags built... plasmids from Penn State Need help picking the right fluorescence microscopy technique to use? Check ...

...feels she was lucky to have been in the right place at the right time when she started her scicomm career...“There were jobs that I didn’t seem to have the right skills for in terms of PR types of things or communications...leverage unpaid work for paid work: “If you use it right, it could help you get your name out for stuff that...

...neutral pH compartment, ­­like the cytosol, to the higher pH of the vacuole. Read on to learn more about ...enzyme. Pho8 phosphatase activity or the molecular weight shift of Pho8 from its uncleaved to cleaved form... dual color-emission biosensor named after the brightly-coloured Australian parrot. It’s comprised of ...Oligomerization Seeing Red: Simple GFP Photoconversion Light Sheet Fluorescence Microscopy Additional Resources...

...cyan, or magenta light. A gRNA with all 3 distinct hairpin sequences produces white light. Created with ...from red, blue, and green channels overlaid on a bright-field image. Image from Ma et al, 2016. Using... A gRNA with all three hairpins generates white light. All 6 gRNAs can be expressed in a single vector...

...plasmid verification, and using NCBI BLAST. Tips for high-throughput plasmid sequencing If you’re like us,...Sanger sequencing is done from glycerol stocks (overnight cultures with 15% glycerol), as opposed to purified...handling of large numbers of samples. When we grow overnight cultures for glycerol stock production and storage...can make your life much easier by selecting the right primer. This often starts by determining what features...

... edition of “What’s New in CRISPR” - where we highlight a few of the newest CRISPR plasmids available ...cells. The “V4” version (named CasMINI) had the highest activity for gene activation  and base editing,... not modify the Cas12f1 enzyme. The result is a highly efficient Cas system with very compact genome editors...

...RNA! Double stranded? Well, that’s just for DNA – right? RNA performs almost all of its biological functions...replication process. dsDNA viruses yield dsRNA through a slightly different mechanism thought to be associated with...always around. R-loops are particularly enriched in highly transcribed regions and repetitive regions, such...RNA:DNA hybrids levels are allowed to reach very high or low levels, possibly due to dysregulation of ...

...result in any cells, or all the colonies screened might not contain the correct plasmid. There’s a lot that... 24 bp with the template and if the sequence has high GC or AT content, increase the overlapping region...Full lawn of cells. The antibiotic in your plate might not work. Check the plates! Streak on them an E....t get any positive clone after these steps, you might want to reconsider your approach, and try a different...

...interactions by cross-linking RNAs and proteins with UV light, followed by an antibody pulldown of a protein of...for Affinity Purification (CRAPome), to identify highly biotinylated background proteins reported by other...and identified QKI, an RNA-binding protein that’s highly expressed in neural progenitor cells. Knock-down...Response Element (IRE) RNA motif of the ferritin light chain RNA transcript. IRE motifs are bound by IRE...

...initiation sites of at least two genes with high expression and high cell-type specificity.  Strategy #3 (ProC...promoter designs, promoters from strategy #4 had the highest rate of targeted expression. Surprisingly, <1% ...tracing experiments: While synthetic promoters with high rates of labeling are ideal of gene therapy applications...

...acknowledgements sections. Some other additions you might wish to include are a references section, contact...navigate the poster from column to column left-to-right, which is the natural way that people expect to ...illustrations above) so that the color fades to a lighter or darker version of itself from the top of the...sure that the color of your font is compatible. Use high-contrast between these colors so that your text ...

...systems Gamma-retroviral vectors have a relatively high transduction efficiency, but in contrast to lentiviral...small-molecule libraries and can be combined with high-throughput sequencing to perform multiplexed analysis... cassette. It can be used to produce a ~10-fold higher titer of lentiviral particles than the original...lentivirus provides significant genetic power and allows high throughput genetic modulation studies in mammalian...

...NGG) in both distribution and GC-content, which might improve editing in AT-rich genomes. Additionally...editing efficiency for the Cpf1 orthologs was only slightly lower than that of the widely used SpCas9 and ...that AsCpf1’s off-target rate is similar to that of high fidelity Cas9s eSpCas9 and SpCas9-HF1. Both AsCpf1... cleaves relatively far away from the PAM, NHEJ might retain the PAM site. Therefore, if HDR does not ...

...elements. A high affinity terminator may be desired for multi-cistronic constructs where high termination...terminators, such as T7 and rrnB, as well as engineered high-efficiency terminators such as T0. Rho-independent...poly-uracil tract as shown in the figure to the right. The tertiary structure of the hairpin-DNA complex...

...RNA. Measuring p24 levels in a viral prep is straightforward and rapid as there are several commercially...Studies have found that values can be 10-1000 fold higher than for functional assays depending on the vector... collected lentiviral prep may be significantly higher than the same prep that has been stored in the ...routinely produce virus may want to choose more straightforward approaches such as the FACS-based or colony-forming...

...Maya Kostman for the IGI.     Here are some highlights of the plasmids shared over the last year and... more tools, head over to this blog post that highlights five different methods to use CRISPR to detect...

...location bind to Cas9 with canonical sgRNA structure (right). Created with BioRender.com. What to edit The...3 or more mismatched bases can often evade MMR (right). Created with BioRender.com. For specific prime...prime editors bound to sgRNAs only drive nicking (right). Start with nicking sgRNAs ~50 bp from the prime...2024). Figure 9: epegRNAs (left) and PE7 (right) offer two different strategies for protecting pegRNAs...

...genome is made of protein-coding genes, and you might be wondering what the rest of our genome could possibly...non-coding RNAs! You probably didn’t see that coming, right? Non-coding RNAs (ncRNAs) are transcribed but do...versatile option. It can be used to target lncRNAs with high specificity, particularly in applications requiring...Hazan J, Bester AC. CRISPR-Based Approaches for the High-Throughput Characterization of Long Non-Coding RNAs...

...just for lab specimens — there is evidence that lightning can also promote gene transfer in soil bacteria...microbial offense One way to triumph over your neighbors is to eliminate the competition, and Penicillium...specialized degradative capabilities are often tightly regulated to be active only in the presence of ...Simonet, P. (2001). Laboratory-scale evidence for lightning-mediated gene transfer in soil. Applied and Environmental...

...If you’ve ever been looking for just the right CRISPR vectors on Addgene and found instead ones that ...Doench reports even the greenest members of his lab, high school interns, were able to use Fragmid to design...making it easier to get the vectors that are just right for your experiment. As always, we recommend checking...components by clicking the blue text in the upper right-hand corner.      Step 3: Reviewing the ...

...the Mobile CRISPRi cassette flanked by left and right Tn7 end sequences. Donor bacteria with a chromosomal...scheme. Mobile-CRISPRi integrates downstream of the highly conserved glutamine-fructose-6-phosphate aminotransferase...cell, the CRISPRi DNA (purple) flanked by left and right Tn7 end sequences (green) integrates downstream ...mirabilis, had only ~1 transfer per 1,000,000 colonies. Higher transfer rates are better suited for genome-wide...