...interneurons using a viral approach Genetically modified organisms like transgenic mice are important tools...efficiency. To further improve specificity, they purified HF-BE3 protein for delivery in ribonucleoprotein...comprised of 2 plasmids, pPSU1 and pPSU2, that can be amplified affordably in bacteria. They are compatible with...strain and contains knockouts for all 3984 genes identified as non-essential. The knockouts replace entire...the arrayed knockdown library targets 289 genes identified or proposed to be essential in B. subtilis 168...systems, which include S. pyogenes (Spy) Cas9, been identified. In Rauch, et al. 2016 the Joseph Bondy-Denomy...inhibit CRISPR function. These Lmo proteins were identified using a bioinformatics approach examining genomes...

...for “technical or product support specialist,” “field application scientist,” or “scientific or technical...career, promote career growth, and/or switch your field of work, both in academia and in industry. Advancing...Advancing along a career path in these two fields looks different however. In industry the career ladder ...Working on a variety of projects and in different fields of research is one of the best parts of my career... with a master’s. While it's possible to change fields of research with a PhD, a master’s is less specialized...

...biomedical research". The family entered the biomedical field’s spotlight after researchers discovered heavy-chain...fragment was exciting, and it wasn’t long until the field was off and running with camelid-derived single-...out of reach and unappealing to many labs), the field is moving towards using synthetic single-domain ...al., 2021).      Figure 2: Genetically modified mice offer an alternative method to producing ... et al., 2021.   The single-domain antibody field has come a long way since the serendipitous discovery...

...understand where the field is today? Have you talked to veterans in the lab or the field? What are possible...paper look like? What will be the impact on your field? You want to choose a research question that people...approach with another in a way not done before in your field?” He points out to “not be shackled by your own ...end point of training is not to become a pro in a field, but to be trained to think and perform like a scientist...

...particularly suitable for in vivo screens, where CRISPR modified cells are transferred into an animal and the behavior...Antibodies could further be used to interfere with the identified surface proteins, and such antibodies could thus... genes were included, and how many gRNAs were identified for each gene. This file can be used to rapidly...choice using the adapter sequences that you have specified.   Run your screen! Final note I am very grateful.... Ma, Hongming, et al. "A CRISPR-based screen identifies genes essential for West-Nile-virus-induced cell...Konstantinos, et al. "A CRISPR dropout screen identifies genetic vulnerabilities and therapeutic targets...

...TANGO) (Kroeze et al., 2015). PRESTO-TANGO uses a modified Tango beta-arrestin recruitment assay to measure...Roth lab showed that this platform accurately identified 120 known GPCR targets demonstrating its potential...proteins near the GPCR. These proteins are then identified with mass spec. Using this technique termed “...optogenetics but instead of light, these genetically modified receptors are activated by small molecules to ...exhibited off-target effects thus scientists then modified some GPCRs further to respond to specific, pharmacologically-inert...

...the depositor field will rank higher than matches of the same name in the publication field. Suggestions... the different usage patterns the larger team identified. The basic function of a search engine is to ...through our educational resources. Filters We identified the most useful kinds of information per resource...words, phrases, partial phrases, or numerical identifiers. We also combine the different terms in a query...

...would have simplified the first weeks or months when I was learning the terminology of a new field. Annotate...collaborations with other scientists within and outside your field, and any number of interactions with professionals...towards solving this problem by 1) making sure we qualified these words when we discussed them in meetings...everyone in the lab to be using words for their field or technology in a consistent way. Just creating...

...nice way to see growth or decline of interest in a field over time. Creating an alert for a search term helps...people are excited about. Following scientists in fields outside your main research focus lets you keep ...universities often tweet about publications in the fields of research they support. Monitoring these types...papers too. You can follow journals related to your field of research or larger journals that publish about... Staying up-to-date on new publications in your field. Feeds are constantly updated, so you’ll never miss...

... in vivo applications, you can consider using unpurified AAV from the supernatant (Goodwin et al., 2020...virions from cells and supernatant then need to be purified by density gradient centrifugation (e.g., iodixanol... Titering Once you’ve produced, harvested, and purified your rAAV, you’ll need to determine the titer ... Transgene Expression in Cultured Cells Using Unpurified Recombinant Adeno-Associated Viral Vectors [Preprint...Y., … Golde, T. E. (2020). Utilizing minimally purified secreted rAAV for rapid and cost-effective manipulation...

...double stranded break (DSB) at a genomic site specified by a guide RNA (gRNA) sequence. Users can design...remains challenging to screen for the genetically modified strains that contain your newly tagged gene. Most...simultaneously introduce a selection marker for screening modified strains. With SapTrap, the user first designs ...multiple target sites. SapTrap vectors can be easily modified to tag hundreds of genes, generating powerful ...localization and activity, CRISPR tagging systems can be modified to assay additional protein characteristics. For...

...withstand changes in temperature. Taq was first identified in the late 1960s during research at hot springs...the reaction is compared to a standard curve of amplified DNA of a known starting concentration. The cycle...reverse transcription and the cDNA is subsequently quantified by qPCR. qPCR is used in a variety of applications... is a method in which multiple targets can be amplified in a single PCR experiment using multiple primers...Single template PCR reaction - one template is amplified using several forward and reverse primer sets....

...AAV-containing cells and media are harvested and purified. In total, this process takes 4-7 days, not counting...exchanged to remove the iodixanol and concentrate the purified virus. This process can be completed in one long...solutions were prepared correctly. When aliquoting purified and buffer exchanged virus, remember to make one...titration by qPCR Overview: AAV titering by qPCR quantifies the number of genome-containing viral particles... primer/probe set can be used and samples are quantified by comparing to an AAV standard curve, which ...

...It was by serendipity that I got into the field of gene therapy, more specifically AAV-based retinal ...trial gone horribly wrong. It brought the whole field to a grinding stop and only now, 2 decades later... work and a little bit of luck A pioneer in the field, Dr. Jean Bennett started working on ways to deliver...Gene therapy was in its infancy, but the eye was identified early on as an ideal target. It is an easily ...have broad tropism in the eye. AAV-2, the first identified serotype and the one used in Luxturna™, can efficiently...

...studied during graduate school. As a stable, easily purified protein with easily measured activity, MDH is ...Provost’s CURE prepare to analyze their recently purified MDH to study the impact of phosphorylation on ... genuine interest to other researchers in their field. Indeed, sharing results with the broader scientific... original and recombinant enzymes in bacteria, purified them to near homogeneity using a 6XHis tag, and...B. 2020. Mentorship and protégé success in STEM fields. Proceedings of the National Academy of Sciences...

...major players in the CRISPR field. Just a year after Feng Zhang’s lab identified Cas13a (C2c2) (Abudayyeh...opposite the target A to be edited. This change specifies the edit to be made when multiple As are present... see if they could shrink the construct. They identified ADAR2DD(delta984-1090), which decreases the REPAIRv1...the authors performed rational mutagenesis and identified that a S375A mutation in RESCUE (named RESCUE-S...

...has been a long and winding path for many in the field. The vast majority of grant professionals didn’t...and by finding our individual niches within the field of grants. What is a grant professional?  In the...grants has fostered the demand for experts in the field of grants. It is no surprise that the U.S. Bureau...less than 400 international Grant Professionals certified through the Grant Professionals Certification ...by the Grant Professionals Association, and a Certified Social Media Strategist through the National Institute...

...that MPG could be engineered to act directly on unmodified G bases, resulting in a G → Y (Y = T or C) base...thymine Since an enzyme that excises inosine can be modified to target guanosine instead, perhaps analogous... et al., 2024). In each case, a UNG enzyme was modified to recognize T and/or C substrates. The HuidaGene...protein engineering going on in the base editing field! From CRISPR-free approaches to mitochondrial base...more diverse. We can’t wait to see what tools the field will come up with next! References and Resources...

... Cas9, Cas9 modified 9/12 known off-targets with a 14% indel rate, while ABE7.10 modified only 4/12 off-targets...target base for editing within the editing window specified by the Cas protein These elements were the starting...of dCas9. The resulting editor, BE3, nicks the unmodified DNA strand so that it appears “newly synthesized...efficiency. To further improve specificity, they purified HF-BE3 protein for delivery in ribonucleoprotein... and that they are expressed well. The Liu lab modified the nuclear localization signals and codon usage...

...isolation of modified cells; and (5) is compatible with the introduction of unique molecular identifiers, which... In vitro CRISPR screens in immune cells have identified many putative regulators of immunity. However... of the pRDA_526 vector. Moreover, the screen identified putative novel regulators of CD8+ T cell responses...