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...cells are a great option for viral production, particularly for labs that already do mammalian cell culture...for the generation of viruses, though they are a popular choice for protein production. The most commonly...infect insects. This is what makes insect cells a popular choice for protein production. AAV and lentiviruses...systems. Baculovirus transfer plasmids have a much larger packaging capacity — up to 38 kb! Generally, insect...in between passages to eliminate dead cells and cellular debris. While adding time (about three to four...supernatant in a centrifuge to remove any unwanted cellular debris, as you would when collecting a lentivirus...

...depositing lab. There may be other plasmids with similar names in the collection, but you can use this name...some of Addgene’s plasmids is an indication of popularity. A yellow flame indicates that a plasmid has ... on our website, one of the available filters popularity (e.g. only showing those plasmids which have ... than 100 times).     I guess it's cool to be popular, but how do I know if a plasmid is actually useful...If the depositor has indicated that there is a particular mutation found within the gene/insert, this information...means you can see which labs have already used a particular plasmid, and which experimental systems it was...

...specific genes in mammalian cells. Despite the popularity of CRISPR-based knockout/knock-in systems, these...nucleotide sequences are exchanged between two similar or identical molecules of DNA. Gene targeting takes...targeting constructs. In the example shown in Figure 1, large sequences corresponding to exons 1 and 3 of the ... the cells that have randomly integrated all or large portions of the plasmid. Deliver your construct ...insert GFP into the genome, we’d create a plasmid similar to the one shown below, where the sequence of GFP...CRISPR is very good at making knockouts, knocking in large sections of DNA can be more difficult. Addgene depositors...

...role of individual genes or entire pathways in a particular disease process. While CRISPR is certainly not... mutant cells to identify genes involved in a particular phenotype. CRISPR can be readily scaled up for...fluorescence-based cell sorting and screened for a particular phenotype. For example, mutant cells can be used...cell types that are of low abundance are not particularly well suited for genome-wide screening. Avoid...targeting the same gene can be strong evidence that a particular gene is actually important for a given phenotype...Lentiviral CRISPR Libraries. Current Protocols in Molecular Biology 115: . https://doi.org/10.1002/cpmb.8 ...

...results. The default setting is “Optimize for Highly similar sequences (megablast)”, which is very fast and ...often unreliable, we suggest using the “Somewhat similar sequences (blastn)” program for blastn so that ...your sequencing trace file. Unlike megablast, the regular blastn program uses a smaller word size and lower...protein sequences (nr)” database as it contains the largest number of protein sequences. Just below the BLAST...options and for our suggested blastx customization. Similar to nucleotide sequences, proteins often have repeated...COBALT for aligning multiple protein sequences, particularly for comparing different isoforms. In addition...

...transistor-like digital genetic switches that enable cellular computing. These logic gates built from transcriptors...of promises to not assert property rights for particular engineered genetic sequences; but it’s only the...computer languages are free to use. Of the 15 most popular computer programming languages, 12 out of 15 are...are now available exist within the context of the larger bioeconomy, and that it’s been 40 years since the... on the domestic market generates $300 billion dollars every year. That's two percent of the U.S. gross... we anticipate plugging into the probiotic and cellular therapeutic space. If you were to look forward...

...information while using less resources and are particularly useful when trying to screen a limited number... libraries have been shown to outperform other popular CRISPR libraries (see below). Let’s take a look...Brunello library was more effective than other popular CRISPR libraries in the depletion of essential ...discriminate between non-essential and essential genes similarly to the Brunello knock-out library while mitigating...between 150-75 nucleotides upstream of the TSS. Similar to the Brunello and Dolcetto library, sgRNAs were...about the advantages of pooled CRISPR libraries for large-scale functional screens Read this blog post about...

... Boyden Lab, ExM has been steadily growing in popularity with an abundance of protocols and examples of...not something you find in the average cell and molecular biology lab, and EM is limited in the number of...provides new insights into the cytoskeleton of malaria parasites including the conservation of a conoid..., Mohar B, Eddison M, et al (2022) Visualizing cellular and tissue ultrastructure using Ten-fold Robust... Zwettler FU, Guennec ML, et al (2019) Imaging cellular ultrastructures using expansion microscopy (U-...Wijesekara P, et al (2023) Magnify is a universal molecular anchoring strategy for expansion microscopy. Nat...

...Interestingly, he also adds to the disproportionately large number of honorees to have trained under a previous...fortunate enough to apprentice with top scientists to larger groups of trainees at an earlier point in their...protein with easily measured activity, MDH is particularly amenable to in-class labs. Provost ultimately... at the American Society for Biochemistry and Molecular Biology Annual Meeting. Some of my students published...and green fluorescent protein. Biochemistry and Molecular Biology Education. 38(5):317-323. Knutson, K.,...dehydrogenase as a research model. Biochemistry and Molecular Biology Education. 38(5):324-329. Kowalski, J....

...specializes in developing novel technologies to analyze molecular mechanisms of genome functions such as epigenetic...transcription and epigenetic regulation. To elucidate molecular mechanisms of regulation of genome functions, ...isolate specific genomic regions while retaining molecular interactions. For this purpose, we developed the...merit of depositing plasmids to Addgene has been spectacular. We've got more than one hundred requests, and...been very difficult for us to respond to such a large number of requests by ourselves without Addgene.... experiences in research labs. What are some similarities and differences you have observed as you have...

... the dsRed protein isolated from Discosoma sp. Similar work has been done to expand the red-fluorescent...fluorescent proteins have become a mainstay in molecular biology. Scientists can easily utilize GFP-containing...biosensors has been designed to detect a variety of intracellular conditions, including ion (such as Ca2+) concentrations...can use light to detect, measure, and control molecular signals, cells, and groups of cells in order to...applications for GFP. It has also been used to identify particular cell populations in drug screens, to visualize...strand break repair, and to label pathogenic intracellular microbes to visualize host/pathogen interactions...

... your ocular lens. For example, if you observe something using the 10x objective and your ocular lens ...transmission electron microscopes that allow us to see cellular components with science-fiction levels of detail...are magnification (the ability to make an image larger) and resolution (the ability to distinguish between...Lenses Focus Knobs (Fine and Coarse) Nosepiece Arm Ocular Lens (Eyepiece) The route the light follows from...objective lens which magnifies the image, then the ocular lens, where the image is further magnified before...objective magnifies it 10 times, and so on. The ocular lens also provides magnification and the power ...diaphragm and use the coarse focus adjustment knob (the larger of the two focus knobs) until you can see the edges...

...commonly used to create functional knockouts or large deletions, the ability to analyze and interpret ...which of the multiplexed sgRNA was involved in a particular edit, and which type of edit was produced.   ...characterizes the sequence and abundance of each particular indel.   For a complete guide to ICE analysis...Typically, a yellow check mark indicates that a particular parameter was automatically adjusted to generate...+F or Command+F functionality to search for a particular sequence or name. You can perform more in-depth...Contributions, Indel Distribution and Traces - provides particular details about the indel profile of the edited...

...Bird A (2020) The Molecular Basis of MeCP2 Function in the Brain. Journal of Molecular Biology 432:1602... consolidated and centralized a collection of molecular biology tools so that both veteran and new Rett...severity. This exciting finding injected hope that a similar reversal may be possible for children and adults...RNA-mediated correction of MECP2 mutant alleles, in particular, have shown exciting and promising results for...Apparent Toxicity after Intracisternal Delivery. Molecular Therapy - Methods & Clinical Development 5:106...

...deliver larger cargo such as base editors and prime editors. While using AAV to deliver large transgenes...Kenkel.  CRISPR genome editing has quickly become a popular system for in vitro and germline genome editing...in mammalian cells. SaCas9 produced indels at a similar efficiency to SpCas9, leading the group to focus... with split AAVs. In general split AAVs break a large transgene in two pieces and package each piece into...two proteins, mediates protein trans-splicing, similar to an intron in pre-mRNA splicing. In 2016, Fine...

...two independent groups, led by Jack Szostak and Larry Gold, developed methods for selecting and amplifying...metal ion, or fold to incorporate themselves into a larger target, such as a protein. Such flexibility makes... Design and selection for targets can be done largely in vitro; Easy and reproducible commercial synthesis...advantages of oligonucleotides over proteins is their modularity, which is lost as soon as the aptamer is bound... Assays that require recognition of ions or particularly small molecules that do not generate an immune...Central PMCID: PMC4071757. 6. Hong, He, et al. “Molecular Imaging with Nucleic Acid Aptamers.” Curr Med ...

...your science. While Reddit isn’t for everyone, particularly if you’re more interested in selling a product...topic”) which are essentially lists of forums on particular topics. Volunteer moderators set the rules for... and, at least sometimes, civil. r/Science in particular hosts a variety of conversations with people ...can ask designated scientists anything about a particular topic. Given Joanne’s expertise in science careers...Reddit. Here are some reasons why: 1. Reddit has a large and active community of people interested in science... moderators did not delete or suppress were the large number of scientist posters with serious career ...

...and its family members serve as the cellular receptors for vesicular stomatitis virus.” Proceedings of ... can persist undetected; unlike infections with larger microbes such as yeast, fungi, or bacteria, mycoplasma... medium. Daily medium exchanges help alleviate cellular stresses associated with pH changes and resource...Detection of mycoplasma contaminations.” Methods in Molecular Biology 946 (2013). 1-13. Pubmed PMID: 23179822...lentiviral vectors: effect of sera and polycations.” Molecular Biotechnology 53.3 (2013). 308-14. Pubmed PMID...

...behind the cytotoxicity can vary. For example, the popular VSV-G envelope protein causes cells that it infects... were produced in. For example, production of vesicular stomatitis virus, HIV-2, and human foamy virus...intact SARS-CoV-2 virus. Scientists have been using similar pseudotyping methods with other viruses in a BSL...envelope protein for pseudotyping is VSV-G, or vesicular stomatitis virus G protein. VSV-G is a trimeric...in Human Cells Are Inactivated by Human Serum. Molecular Therapy 2:218–222 . https://doi.org/10.1006/mthe...nervous system after peripheral delivery. Human Molecular Genetics 10:2109–2121 . https://doi.org/10.1093...

..., to search the literature, and to synthesize a large volume of information: basically, a scientist’s ...colleague, or your adviser was invited to write on a particular topic. Alternatively, maybe you’re researching...helps you later on to remember where you read particular studies or experiments. Later, you can easily... searched for recent research papers on these particular themes, for seminal papers on these themes, and...also did not limit myself to the well-cited or popular papers, but looked for papers from a wide range...present it all in one place to get a sense of the similarities and differences among proteins in the class....