...problems. Rapidly dividing cells initiate genome replication more than once per cell division and thus, genes...genes closer to the origin of replication will be overrepresented compared to those further away. As a...cells are dividing rapidly, multiple rounds of replication are initiated before the cell has divided. This...maximal gene expression near genomic origins of replication, the stabilized promoters created by Segall-Shapiro...the control of IPTG. Because RepA is a plasmid replication initiator protein, they could alter copy number...dosage in E. coli using DIAL strains." Journal of biological engineering 5.1 (2011): 10. PubMed PMID: 21787416...

...days prior to the implementation date, our data indicated that, while 69% of all postdocs were expected ... department at the University of Massachusetts Medical School. We are working with postdocs at other institutions...sent, with the lack of transparency and poor communication of their plans (many postdocs - and in some ...to reach out to institutions with a statement indicating that we encourage them to keep raising salaries... December 1st. Salary inconsistencies across geographical regions When we originally started gathering...changes in science. Follow him on Twitter @BiophysicalFrog and Future of Research @FORsymp.   Additional...

...editing performance at therapeutically relevant gene targets and expand the application to the possible treatment...AsCas12f1. They found that this system, without any modifications, is effective in genome editing in both bacteria...ribonucleoprotein, and AAV.  Read the paper in Nature Chemical Biology Find the AsCas12f1 plasmids   Selective... Mike Lacy The retroviral-like protein PEG10 specifically binds its own mRNA to facilitate its packaging... the prime editor, yielding PEmax, to work synergistically with epegRNAs and PE4 and PE5. These enhanced...

... audience and convey your ideas clearly and strategically is powerful in almost any career. It allows ...interview better! As a scientist turned science communicator, I’ve done a lot of public speaking, and I'm... the open windows in the middle of your talk. Technical difficulties turn your slide deck into a chalk...everyone! Everybody makes mistakes, encounters technical difficulties, and runs into unexpected stumbling..., to a group of peers who are there to provide critical feedback, often with a semi-international audience...your own native language. And, at least in the biomedical field, with very little text on the slides and...

...composed of four peptide chains (two identical heavy chains and two identical light chains) each with a constant...antibodies means that it is relatively easy to physically divide up the different regions, both through... as secondary antibodies, have much broader applicability. It is fairly easy to find these in a Fab fragment...fragments described above can be used for similar applications and have similar advantages, though with some...David vs. Goliath: The Structure, Function, and Clinical Prospects of Antibody Fragments. Antibodies 8(...

...and on self-propagation by reproduction. Both physiological processes are controlled by deliberate and compulsory...receptors (NRs), their coregulators, and their physiological endocrine ligands. Nuclear receptors and their...murine members. NRs permit the integration and communication of such signals between central and peripheral...circadian regulation, as well as in important physiological states such as adipogenesis and macrophage activation... activation. Our mass spectrometry-affinity purification proteomic profiling of coregulators in HeLa cells...widely used today.  The sheer abundance of these critical research tools is a double-edged sword, however...

...prevent the formation of replication competent lentivirus (RCL). Replication competence can arise from...end-users, size of the transgene, and downstream applications should all be considered when choosing the correct...unconcentrated prep. For example, at Addgene we typically concentrate 100-fold meaning that a 10 cm2 dish... as resources available, cost and downstream applications. For example, while the PEG precipitation method...lentivirus is the cell line that is used for titering. Typically researchers will choose a relatively permissive...

.... When an ELISA kit is ordered, the wells are typically already coated with the antibody. Then, you add...process is the addition of a detection antibody (typically a monoclonal antibody) that also recognizes the...the most common type of competitive ELISA is a modification of the sandwich ELISA as described here, the...ELISA, the only difference being the additional amplification step, which helps to visualize low-abundance...concentrations of your protein of interest. The optical density (OD) - the measure of light absorbed by...concentration of any experimental wells in that range of optical densities. If your standard curve does not have...

... for PAM-less editing   Guides for CRISPR are typically selected by the ‘quality’ of the gRNA sequence...location of the cut and subsequent edit. In these applications, location takes priority over cutting efficiency...the Liu lab through phage-assisted evolution, specifically with the goal of generating Cas enzymes with...compatibility. This alternative PAM requirement adds a significant amount of flexibility compared to a strict single-compatibility...in double stranded cleavage and base editing applications, with high editing capacity in both contexts...increases the flexibility of gRNA placement significantly over SpCas9 and more so than Cas9s with an NG...

...targeting anywhere in an exon. But things get more complicated if you’d like to use homology directed repair...edits or knock-ins. Editing efficiency decreases drastically once your cut site is more than 10 bp away from...They created a selection phage (SP) to carry catalytically dead SpCas9 (dCas9) fused to the bacterial polymerase...loci displayed a broad range of PAM sequences, indicating they may be less predictable than SpCas9 off-... that we have only scratched the surface of modifications that can be made to Cas9. It is a pleasant surprise...editing rates. This PAM flexibility should expand applications that require very specific targeting, like homology...