...in bacteria, most CRISPR-based genome engineering has taken place in other organisms. In many bacteria,... targets cells in which homologous recombination has not occurred. This powerful negative selection system... works similar to the original pCas/pTargetF but has many other advantages including shorter plasmid curing...plasmids Noting that the more common spCas9 system has generally low efficiencies or can be proteotoxic ...system can be delivered a number of ways but success has been observed with Phagemids (Selle et al., 2020,...microbiome Beyond the human gut, the Barrick lab has deposited plasmids that can be used to engineer the...

...Henry Bourne, Professor Emeritus at UCSF. Dr. Bourne has some radical (and some not so radical) suggestions...called The Balsa Group. University of Pennsylvania has the Penn Biotech Group and PBG Healthcare Consulting...

...and you need the right experimental materials. Who has time for paperwork? Here’s why a Material Transfer...confidence. Benefits of the UBMTA: Peace of mind Addgene has adopted the terms of the Uniform Biological Material...

... flow rate into the pipette tip. Once the liquid has finished drawing into the pipette tip, lift the pipette...

...effective than both AAV9 and AAV5, which has tropism for lung cells, and has increased overall transduction throughout...this definition: A systemic capsid is a capsid that has been engineered to have enhanced function after intravenous...both). Therefore, a high-efficiency systemic capsid has better efficacy than AAV9 and/or its intermediate...used in LY6A-nonpermissive mouse strains. **AAV1-X1 has not been tested in rats. For more information on ...neurons in the CNS (Chan et al., 2017). AAV-PHP.eB has frequently been used for screening candidate regulatory...depositing manuscript) across the brain. AAV-BI-28 has been tested in C57BL/6J mice and effectively transduces.... 4c, Jang et al., 2023; Brown et al., 2021.) It has a similar transduction rate to AAV-PHP.eB in the ...

... to its unrivaled contrast, fluorescence imaging has emerged as the dominant light microscopy contrast...blue to excite a molecule of GFP. Because red light has approximately half the energy of blue light, two ... be establish for this to happen.Using red light has a second advantage: red light penetrates deeper into...imaging depths closer to 1 mm. Traditionally, 2-photon has been a very slow imaging technique but recent advances... modern explosion in light microscopy techniques has provided biologists with a host of tools and opened...

... cell mobility within these complex environments has never been easy and, for many years, researchers ...Intravital imaging powered by two-photon microscopy has uncovered host-pathogen interactions leading to the...proteins can be used in your experiments, experience has shown that some are more reliable and more efficient...important to finely tune the excitation wavelength as it has been shown that even a small incremental change of...this system called Brainbow, the Ubow mouse strain has been created to fate map Langerhans cells and follicular...

...the Fire-Cas9 Podcast Segment Jerry Crabtree’s lab has developed a new CRISPR/Cas9-based system to study.... While catalytically inactive cas9 (e.g. dCas9) has been fused to a variety of protein modules to repress...synthetic transcription factors, the El-Samad lab has deposited plasmids encoding fluorescent reporters...

...VSV-G that generally mediates broad cell tropism and has a high percentage of neuronal targeting after stereotaxic... damage to the brain tissue. The injection speed has to be slow to allow the viral particles to diffuse...order to minimize the pressure into the tissue, but has to be large enough to target larger areas, typically...

...called the 4D nucleome. Visualizing specific loci has historically been difficult, as techniques like fluorescent...visualize genomic loci in vivo The flexibility of CRISPR has improved our ability to target almost every genomic... system and induce genome editing. Such a system has been previously described for transcriptional activation...

...all four parts of a STAR answer once the interview has started. Instead, I preferred to prepare stories ...stories for an interview, since one story usually has more than one theme and can be used to answer multiple...new idea to the company’s area of expertise. This has potential to make your audience really sit up and...

... a lot quicker to pick up new languages!) Jargon has a place in science communication, but the more you...length of your average PhD to achieve!) Each field has its own framework: a particular way of organizing... amount of knowledge and expertise your audience has is a major factor in how quickly they can understand...

...antibody is biotin-conjugated, streptavidin (which has a high affinity for binding biotin), is conjugated...detecting one protein from the sample. A direct ELISA has many fewer steps than a sandwich or competitive ELISA...linear portion or the OD of your experimental samples has reached 4.0, you can resolve this issue when you ...

...CombiGEM-CRISPR: The marriage of two simple platforms CRISPR has revolutionized how we decode the genome by making...guide RNAs (gRNAs) for genome editing in large-scale has led to the rapid generation of genome-wide gRNA libraries...the users’ specific research questions. CombiGEM has been successfully applied to functionally characterize...

...auditory neuronal circuitry in vivo. A variant of RABV has been developed to only label a neuronal connection...example only dopaminergic neurons, neuronal tracing has the added benefit of only labelling the subset of... regions used. Evidence for retrograde transport has been found for AAV1, -2, -5, -7, -8 (Taymans et al...AAV5 (Aschauer et al., 2013). Anterograde transport has been observed for AAV1, -5, -8 (McFarland et al.,...variability compared to chemical tracers. rAAV2-retro also has lower affinity for heparin, which possibly explains...vectors cover some of these attributes and their use has allowed for the visualization of single connection...intracerebral application, which is more invasive and has a much higher risk of complications. References...

...IgG2a  Rat   Targeting integrins in the lab The Ha and Springer labs recently showed RGD-binding integrins... interpret the roles of individual integrins. The Ha lab — in collaboration with the Springer lab, the...Leucine-Valine-Aspartate-Proline (LVDP) motif. The Ha and Springer teams employed IPI synthetic antigen-binding... J., Yan, J., Waterman, C. M., Springer, T. A., & Ha, T. (2022). Single-molecule characterization of subtype-specific...

...in the same field that I do: prostate cancer. She has guided me in my next steps. For example, how to get...to travel abroad, how to write cover letters, and has suggested important things to emphasize in order ...insecurities… all shrink when shared with a mentor who has experienced them before and who can serve as a role...universities and future generations of professionals has made possible a significant qualitative and quantitative...

...lentiviral service is going strong, the AAV service has shown incredible growth this year. pAAV-hSyn-DIO-...pmSyn1-EBFP-Cre from Hongkui Zeng’s lab. This AAV has had over 150 orders since coming online! pmSyn1-EBFP-Cre...

...community has always emphasized the need for open science and collaboration. The field already has comprehensive...first virtual organism in a computer.” This project has produced various resources for the worm community...

...the idea that DNA methylation is linked to disease has now become much like the 1980s song “Don’t Stop Believin...none existed. Recently, however, a particular tool has gained widespread popularity – and notoriety – as...may later be excised by Cre recombinase. Addgene has several great options: pLV hUbC-dCas9-T2A-GFP for...