...After several decades of trying to address these challenges, researchers have amassed a considerable molecular... overexpress a recombinant protein? It is not generally in a cell’s best interest to overexpress a protein...peptide-based proteosome inhibitors, glutathione S-transferase (GST), which can be fused to recombinant proteins...for affinity and purification An affinity tag, generally a relatively small sequence of amino acids, is...Purification Binding and elution steps use very moderate buffer conditions FLAG DYKDDDD or DYKDDDDK or...interest would eventually get ridiculous (you generally don’t want more tag than protein), 2-3 tags is...IgG-binding domains. TAP has since come to encompass several other tag combinations, though frequently those...

...lines in tissue cultures takes half a year, and generating a transgenic plant can take even longer, a transient...are investigating how the production of cancer therapeutic drugs, vinblastine and vincristine, are regulated...transgenic hairy roots takes at least six months. Generating a transgenic plant takes even longer, is more... that neither a stable cell line or plant is regenerated from the transformed tissue. Gene function is...by working with young seedlings and optimizing several other aspects of the transient transformation procedure...Once we were on the right track, we switched to luciferases to more easily quantify the effect of the different...GUS::tMAS to confirm transformation success. Luciferases are great quantitative reporters and use a substrate...

...composed of actin and myosin which interact with each other to generate forces that cause cells and muscles...signalling pathway, which is a key regulator of all bilateral animal development. SHH plays a key role in proper...into the right and left hemisphere. Addgene has several plasmids that contain SHH, you can find them all...that are necessary for myosin activity and thus interaction with actin in nonmuscle systems in the fly. Spaghetti...other hand, is important in the formation of the visceral mesoderm which differentiates into the muscles...Embryos that lack bagpipe have severely disrupted visceral mesoderm and subsequently the gut does not form...is required for specification of the heart and visceral muscles in Drosophila." Development118.3 (1993...

...directly into at least one and preferably more than one of these five general purpose repositories. RSpace...own pros and cons. All ELNs have the virtue of liberating data from paper into an electronic environment...data out of the ELN; and (2) facilitate inter-operability with other research tools and infrastructure ...of Edinburgh, the University of Goettingen, and several other large research institutions. The following...repositories Over the past decade or so a number of general purpose repositories have been launched and are... associated with publications into one of five general purpose open source data repositories: Dataverse...one foot on the other side of the pond through interacting with colleagues in Edinburgh. You can follow ...

...detected on commercial lateral flow strips, similar to pregnancy tests. With lateral flow strips, the presence... to cleave ssRNA or mRNA. It also exhibits a “collateral effect” of non-specific ssRNA cleavage. How does...sample is first amplified using RPA (recombinase polymerase amplification) or reverse transcriptase (RT)-... for nucleic acid detection almost anywhere as lateral flow strips are easy to transport and work rapidly...diagnostics (e.g. pathogen or virus detection), therapeutics and sensitive genotyping. The beauty of SHERLOCK...Cas12a however recognizes a different PAM site and generates 5’ and 3’ staggered ends after dsDNA breaks. The... DETECTR platform to detect SARS-CoV-2 using a lateral flow strip format. They published this in Nature...

...workflows considerably easier, faster, and more efficient than previous methods. Still, generating reliable...sequence you used. The ICE tool will calculate overall editing efficiency and determine the profiles of...detect or analyze complex edits, such as those generated by multiplexed delivery of multiple sgRNAs. Since...particular parameter was automatically adjusted to generate ICE results. If there are no results or there ... the ICE linear regression is computed during generation of the ICE Score, the Pearson correlation coefficient...around the cut site. It shows, base-by-base, the average amount of signal that disagrees with the reference...sequencing bases downstream of the cut. Summary ICE generates NGS-quality CRISPR editing analysis from Sanger...

... are several factors that may prevent your gene from being up-regulated. For instance, generally, the ...known functions has the potential to be used in therapeutics, as it can increase gene expression at certain...be used to map molecular pathways and genetic interactions. Additionally, large scale CRISPR-based screens...Cas9 cutting activity, off target effects are generally not regarded as being a large problem for Cas9...driving aberrant transcription. That being said, we generally pick guides by putting the promoter of the gene...innovative technologies. To date his group has develop several popular CRISPR-based tools for altering DNA sequence..., Garnett MJ (2019) Prioritization of cancer therapeutic targets using CRISPR–Cas9 screens. Nature 568...

...what online tools people generally like to use to share ideas – with the general goal of seeing if and how...their disciplines. Included in the article is an interactive chart where you can see the yearly interdisciplinarity...abstracts identified. The journal Cell had the largest average view count per video abstract at 2,947 views. The...Gastroenterology (1,046 views) rounded out the top four average video abstract view counts.” For researchers looking...offers a low-barrier opportunity for researchers to leverage the video medium to communicate their research...their contribution. It offers a 5-step process and several examples for creating effective research videos... botanist and oceanographer who has a blog and several videos on sharing your research through video. ...

...them remain uncharacterized. This represents a considerable barrier to furthering research and developing...Cancer Center tackled this problem by developing a moderate throughput functional genomic platform to annotate...over 1,000 mutations, and the group has deposited several hundred plasmids from this study with Addgene. ...research team identified 1,049 perturbations in several genes spanning 33 cancer types. The types of mutations...indels (Ng et al., 2018). From this data, they generated lentiviral constructs using the High-Throughput...cell lines and any mutations that that confer proliferation advantages will be revealed in the absence of...is important for the development of targeted therapeutic interventions for cancer. This screening platform...

... gene expression in yeast Megan McClean’s lab generated an optogenetic toolkit that controls repression...frequency of genome-wide off-target DNA editing. Using several rapid and cost-effective screening methods, David...with reduced Cas9-independent deamination. They generated these editors using the BE4max architecture. The...reported in this study have ~10-100-fold lower average Cas9-independent off-target DNA editing, ~5–50-...off-target DNA editing, and maintain ~50–90% of average on-target DNA-editing levels relative to BE4max...division, and cellular differentiation, but it takes several weeks to delete or deplete individual genes. Michael...depending on transgene design and delivery. They generated hPSC lines with AAVS1-integrated, inducible, and...