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...simultaneously. Image from Li et al., 2021.   The updates involved the following changes. In pCas9, we 1)...the ccdB gene containing BsaI restriction enzyme sites at its ends to generate pEcgRNA. The ccdB serves...32 hours instead of ~60 hours. We successfully tested the pEcCas/pEcgRNA system in 4 E. coli K-12 strains...strain, and 1 Tatumella citrea, with a total of 70 sites edited. We optimized the strategy for plasmid curing... into crRNA arrays targeting multiple different sites in E. coli. pCutamp cures the plasmids by targeting...Chinese Academy of Sciences. He joined Shanghai Institutes for Biological Sciences in 2000 and became a ...

...instance, GFP may occlude catalytic sites or obstruct binding sites or motifs that are necessary for post-translational...avidity because it doubles the number of binding sites per unit.  There are several ways to measure stickiness...Where should the fluorescent protein be inserted? Sites that are less likely to disrupt the structure of...this is not conclusive evidence that the fusion operates in the same way as the native protein. Although...PMC4927352. Janetopoulos, Chris, Tian Jin, and Peter Devreotes. "Receptor-mediated activation of heterotrimeric...

...   This system is distinct from those used in eukaryotes in that CRISPR isn’t the primary editing force... similarly to CRISPR/Cas9 systems adapted for eukaryotes in that Cas9-mediated cleavage induces HDR to...design more familiar to those using CRISPR in eukaryotes (Qi et al., 2013). As in other systems, catalytically...either Gram-negative bacteria or Gram-positive firmicutes, to bring CRISPRi to a much wider swath of bacterial...Pseudomonas syringae. These vectors contain BsaI sites for easy cloning of a gRNA of interest. Cytidine...organisms in the genus. Work from the Sun lab demonstrates the utility of cytidine and adenine base editors...variant of the BE3 that further improves editing rates. References Bikard D, Euler CW, Jiang W, Nussenzweig...

...follow #SfN14 on Twitter for live tweeting updates and the latest news from attendees. And for those of you...students and postdocs, which accounts for many, many lattes and espressos. The energy boost is well needed ...symposium discussing progress made in reprogramming astrocytes to functional neurons in vivo. And I’ve seen ...cup and get back to our booth, I will have more updates coming soon. Stop by booth #3431 if you’re at #...

...each tagged protein must be tested carefully to ensure it retains the attributes of the native protein. Since...accessibility to an antibody. From their initial pool of candidates, they selected 12 candidate protein domains from... from non-vertebrate systems. Preliminary testing indicated that inntags IT5, IT6, and IT10 expressed ...expressed well in cells and did not form undesirable aggregates. Using budding yeast as a model system, Georgieva...

...efficiently transfer a range of protein sizes in minutes without any transfer buffer. These systems, however...Different immunoassays detect proteins in different states. For example, in immunoprecipitation (IP) the antibody...physiologically relevant levels. Some vendors will also test their antibody in a variety of tissues to demonstrate... methylation and ubiquitination. For example, to test if a protein is phosphorylated following a specific...for subcellular fractionations, a procedure that isolates proteins from different cellular compartments ...antibodies that are validated for your application and test the antibody in your specific system. If you found...

...Dephosphorylation Rates: FRET-based reporters can have slower dephosphorylation rates than KTRs which is...import rates can influence KTR translocation. Work from the Covert lab suggests that these rates have negligible...negligible effect on KTRs, however nuclear translocation rates are another variable to consider when designing ...

... frequently used elements in plant expression cassettes: plant promoters, N-terminal tags, coding sequences...C-terminal tags, plant terminators and resistance cassettes for selection of transgenic plants. The complete... gives the option to stack several expression cassettes onto a single T-DNA in subsequent rounds, dramatically...always have the same recognition site but distinct sites where the enzyme will cut. You only need a single...available here.) Although Lohmann said they’ve only tested GreenGate in Arabidopsis, it should in principle...

..."The greatest good you can do for another is not just to share your riches but to reveal to him his own...characteristics of a good mentor: Yoda image courtesy of jorenl/Vecteezy.com Helps set concrete... helps you reach your goals Listens well and communicates clearly and openly Gives honest feedback without...to take risks A great Mentor is often one that devotes time and energy to thinking about how to be a great...professor at a research university in the northeast, writes a thoughtful treatise on the kind of advisor/mentor...

...our in-stock viral preps: ddPCR, purity testing, sterility testing, and viral genomic sequencing to confirm...newsletter for updates on all things Addgene; or subscribe to the Addgene blog for updates and educational...that Addgene cannot perform functional or in vivo testing. We recommend you check the original paper of your...

...induce recombination at pseudo- or cryptic loxP sites (estimated to occur at a frequency of 1.2 per megabase...used successfully in Drosophila, so Rubin sought to test the suitability of four additional yeast recombinases...recombination site for each enzyme, were generated to test recombination efficiency. When expressed in neuronal...each recombinase and reporter construct pair were tested for excision efficiency. B2 and R showed partial...doi:10.1007/s004270100167. PubMed. Cryptic loxP sites in mammalian genomes: genome-wide distribution and...

...mismatched recognition sites, its presence induces first opsin inversion, and then lox sites excision, therefore...Mei and Zhang, 2012). For example, blue light activates channelrhodopsins that, in turn, trigger neural...neural excitation, whereas yellow light activates halorhodopsins to silence neuronal activity (Figure 1) ...induce DNA recombination at defined recognition sites (i.e., loxP and FRT, respectively) (Abremski and...floxed”) by two sets of incompatible recognition sites (e.g., loxP and lox511) in opposite orientations...

...from the NIH, in the United States) Strong letters of reference that attest to your talent and expertise...fellowship lasts five years. Most universities and institutes annually advertise for Group Leader positions... a postdoctoral mentor. A good mentor typically writes the grants that fund your research, guides the ...Here is my personal list:   Exercise - nothing dissipates stress and improves your perspective like intense... Jennifer Lippincott-Schwartz at the National Institutes of Health, was an Associate Professor at Albert...

...also write test question banks, case studies, and experiments for labs, in addition to testing those experiments... at eScience Labs, an educational company that creates hands-on science lab kits and digital curricula.... [We ask them,] 'Can you read this lab. Can you test this experiment? Can you follow and understand the...working with students and faculty, Caitlin also collaborates with the sales and operations departments at...

...Harrington et al tested GeoCas9’s love of heat in two ways. The first set of experiments tested the ability...following heat treatment. The second set of experiments tested the ability of GeoCas9 RNPs to cleave substrate...genomic edits in mammalian cells. Harrington et al tested GeoCas9’s editing skills by transfecting HEK293...thermostability also had a longer lifetime in human blood. To test if this was also true for GeoCas9, Harrington et...

...recombination sites. To introduce a gene of interest into the destination plasmid, recombination sites must first... fragment within the gene or loss of the gene alleviates its toxic effect. Negative selection is sometimes...and integration of the gene of interest at these sites in the plasmid excises the ccdB gene and the cells...gene is used for negative selection on 5% sucrose plates. Exchange of the sacB gene for the gene of interest...

...and Rep40) Rep proteins. Alternative splicing generates the different C-terminal regions of the Rep proteins...key role in AAV genome replication. Replication initiates from the AAV genome ITRs on the transfer plasmid...in the ITRs, uses its endonuclease activity to generates a ssDNA substrate which is then nicked by its ...Alternative splicing and differing translational start sites generate the three VP proteins. The three VP proteins...alternative open reading frame (ORF). While AAP promotes virion assembly, the role of MAAP has not been...

...drop off the box at any location where the United States Postal Service picks up mail. NEB actually started...drop off the box at any location where the United States Postal Service picks up mail. Recycle plastic...it has to be packaging (no coated tissue culture plates, for example). Check the link for more details ...back. Be that person! If you use serological pipettes from Corning®, Falcon®, or Axygen®, set up a bin...in sustainability. Sign up for blog posts and updates from Labconscious. If you’re inspired by this...

...talks by prominent scientists, including Nobel Laureates, who speak about their work and current, trending...society currently has 1,283 members, who comprise undergraduates, graduate students, postdocs, faculty members...The society encourages interactions between undergraduates and PhD students so that they can help support...students to attend. It also hosts high level academic debates for its members to become involved with. The society...over 170 members in its first year. The society facilitates opportunities to learn about Synthetic Biology...

...year, scientists hijacked survival tactics of prokaryotes to deliver the technological biological blockbuster...demands of the biotech field, let's look at how prokaryotes originally utilized CRISPR/Cas as a powerful ...experiments exposed S. thermophilus to predatory phages to test if the exogenous DNA would be incorporated into ...these studies led scientists to hypothesize that prokaryotes developed an adaptive immune system that utilized...an interesting "arms race" between phages and prokaryotes; the ultimate victor will be the species who ...