...using bacterial phage vectors with CRISPR/cas9 sensitizing gene cassettes. Thse vectors are engineered to...genes in the bacteria thus making the bacteria sensitive to known antibotics. These might be used on hospital...

...institutions. The conversations often move quickly into sensitive topics and uncomfortable places, from sticky ethical...rapidly changing landscape of genetics and our consistent stream of events mean that there is never a dull...

...lack all viral genes, including those that are responsible for integration into host chromosomes, further...to other gene delivery tools that are under consideration, recent advancements in various AAV technologies...

...in different fields that can provide different insight into an area - not just working within the established...collaborators can also help you get some critical insight into what you're doing wrong. All this helps to...

...strengths and work effectively. Leadership responsibilities Being a leader in an organization, you need... different hats and came to understand the responsibilities that come with leadership. The NPA was a great...

... a company. Revolution Bioengineering is two responsible adults in their 30s who have quit their academic...below. We’re happy to answer questions about this transition (so far) and what we’ve learned.  Thank you ...

... increase efficiency of these systems, though consistent expression can still be a challenge. A multitasking...powerful than it is. The nuclease domain of Cas3 is considered weak, especially compared to the power of Cas9...

...which way your population needs to be shifted, consider your perpendicular lines. In this case, the population...applying very large compensation values (100+), consider what two markers your fluorescent markers are ...

...epitope, a phenomenon known as sequence context sensitivity (Schüchner, et al. 2020). In their study, some... newer Myc-tag antibodies appeared to be less sensitive to sequence context.    So, do Schüchner, et al...

...nuclease-null—in order to effect different perturbations. Consider, for instance, the challenge of attempting to ...alterations can be exploited to quickly and inexpensively generate a potent Cas9-based transcriptional...

...suppressors, and their protein products. Proto-oncogenes consist of a group of genes/proteins that, when expressed...Ian A., Paul D. Lewis, and Carla Mattos. "A comprehensive survey of Ras mutations in cancer." Cancer research...

...normally lies around 7.35-7.4 so mammalian cells are sensitive to pH. Anything between orange and light orange...Additional passages will allow your cells to achieve a consistent growth rate. For every minute spent organizing...

..., cultures treated with a folA gRNA were more sensitive to trimethoprim when compared to control gRNA ...for trimethoprim, there would be no changes in sensitivity. Pooled knockdowns are also possible with Mobile-CRISPRi...

...primarily found in the culture media, you should consider performing PEG precipitation. Instead of sonicating...solutions are carefully layered and then the viral suspension generated during AAV production is overlaid. ...

... 3-D path of the protein’s backbone in three-dimensional space, but their flexible and jiggly nature suggested...structure into experimentally determined electron density, have been known to say something silly like “See...

...can survive positive selection, but it will be sensitive to negative selection. Remove the positive selection...creating knockout animals or cell lines, please consider sharing them by depositing with Addgene! Figures...

... capture large field-of-views (6). For more comprehensive reviews on several other LSFM configurations...Bradke laboratories later developed the 3DISCO (3-dimensional imaging of solvent cleared organs) method of ...

... 10% bleach. Safety Warnings AAV is generally considered biosafety level 1 but may require BSL-2 handling...45° angle. Count to 20 while slowly and gently dispensing the droplets down the side of the tube. Place... 95 10 2 1 Denaturation 95 0.5 2 50 Annealing/Extension 60 1 2 50 Signal Stabilization 98 10 2 1 Hold ... should be close to zero (B08). At Addgene, we consider runs with an NTC > 10 to be invalid. To reduce...

...students. But he also found that his teaching responsibilities left him with limited time for one-on-one ...notes the importance of acknowledging student apprehensions about “failure” (protocols not working, hypotheses...

...connections between biology and ancestral groups or, by extension, race. Many researchers do large-scale CRISPR ...potential for bias but to actively look for and consider bias as a confounding factor when interpreting...