We narrowed to 823 results for: lars

...CRISPR, is turning potential into impact. In molecular and cellular biology, this happens through the development...this information to identify a large variety of related bacteria with similar Cas systems. Some of these ...exclusively on this space — developing tools for cellular targeting. Lately, they’ve been working on two...of Cas7-11 were reported as up to 85%, broadly similar to shRNA and Cas13 systems in a range of mammalian...fused integrase would then integrate into with a large DNA insert – or Programmable Addition via Site-specific...programmed for new genes. In comparison with other large insert techniques like HITI or HDR, which require... and adenovirus templates. The system, which is larger than Cas9, can also be used with mRNA and a synthetic...

... cell types depending on their serotype, are particularly challenging. Their efficacy in any given system...and to verify the quality of the prep. This was a large factor in Addgene’s decision to launch our viral...category, available from Addgene as of 2023. A large percentage of viral vector preps contain biosensors...were distributed in 2023 alone. You’ll also find a large number of requests for vectors in the “controls”... 4: A) Cumulative AAV preps for the seven most popular serotypes available from Addgene from 2016–2023...2023. B) AAV preps for the seven most popular serotypes distributed per year by Addgene from 2016–2023. ...in 2016, and it quickly became one of our most popular ready-to-use viral vectors, making up almost 10%...

...Replication and control of circular bacterial plasmids. Microbiology and Molecular Biology Reviews: MMBR, ...Plasmids 101 Posts Get practical molecular biology help with our Molecular Biology Reference Pages   ...maintaining a stable concentration of plasmid. Similar to replication in E. coli, regulation primarily...replication of the plasmid proceeds (and terminates) similarly to the bacterial chromosome. Since replication... thereby the overall replication of pSC101 (del Solar et al., 1998). If that’s not enough to keep replication...must fold into a “pseudoknot” held together by intramolecular base pairs for the protein to be translated...5213. https://doi.org/10.1093/emboj/17.17.5201 del Solar, G., Giraldo, R., Ruiz-Echevarría, M. J., Espinosa...

...designed to hold large amounts of data. Although it may be useful to selectively bring particular items into... permissions to view and edit to particular groups and/or particular individuals, the issue of third-party...the University of Goettingen, and several other large research institutions. The following graphic depicts...small fraction of all data directly relevant to a particular experiment, series of experiments, or project...Integrations with additional tools are added on a regular basis, so the ecosystem of related tools is always...capture the conversations and associate them with a particular experiment or project thus significantly enhances... Resources on Addgene.org Find protocols of molecular biology techniques Find plasmids at Addgene Make...

...distinguish between two similar types of HPV, HPV16 and HPV18, from human cells, at atomolar levels, within one...platforms to reliably detect nucleic acids at the atomolar level. The Zhang lab has adapted natural RNase...reporter enables SHERLOCK to detect DNA or RNA with atomolar sensitivity and single base pair mismatch specificity...be detected on commercial lateral flow strips, similar to pregnancy tests. With lateral flow strips, the...CRISPR Cas variant that can also cleave dsDNA similarly to Cas9 (Zetsche et al., 2015 ). Cas12a however...DETECTR. How does DETECTR work? DETECTR works similarly to SHERLOCK. Cas12a is targeted to a specific ...nucleic acids in a mixed population for an array of molecular and clinical diagnostic applications. The company...

...also be used to map molecular pathways and genetic interactions. Additionally, large scale CRISPR-based...guides directed towards that gene as there can be a large difference in guide potency. Thus, in cases where...oligo library synthesis. Prior to Cas9 activators, similar tools were made using other DNA binding proteins...makes it much cheaper to use Cas9 activators. This larger scale of activation allows for exploratory-based...given cell type or organism, have been used in a similar manner to Cas9 activators. However, these can be...gene, as, many times, the isoform(s) common to a particular cell type are unknown or not readily available...target effects are generally not regarded as being a large problem for Cas9 activators. This is believed to...

...smn1 gene is known to cause spinal muscular atrophy, the molecular mechanisms behind the disease are unclear...devastating afflictions. Elia Di Schiavi - Spinal muscular atrophy Dr. Elia Di Schiavi is a Research Scientist...Bioresources in Naples, Italy. His group works on spinal muscular atrophy, a disease in which disruption of the ...cells. With a more concrete understanding of the molecular biology behind SMN1 and its interaction partners...thereby restore motor neuron function in spinal muscular atrophy patients. Noushin Nabavi - Peritoneal ...mesothelioma subsets. Peritoneal mesothelioma in particular takes root in the tissues surrounding the abdomen...Systematic review of incidence and prevalence of spinal muscular atrophy (SMA)." European Journal of Paediatric...

...miRFPnanos9 do not yield to (m)iRFPs in cellular and molecular brightness (Table 1).  NIR reporters and...). Modern state-of-the-art NIR FPs can be used similar to GFP-like probes by supplying a single gene to...brightness of NIR FPs in cells, in addition to their molecular brightness (a product of the fluorescence quantum...biosensors can be imaged at spatial scales from sub-cellular to whole-body in small animals, such as mice (...phytochrome (BphPs) photoreceptors have higher apparent cellular brightness than previously reported widely used...and biosensors NIR reporters can tell us about cellular events via  fluorescence changes occurring due...FP Ex, nm Em, nm ECa, M-1cm-1 QYb, %   Molecular brightness vs. iRFP713, % Oligomeric state Photo-stabilityc...

...disulfide bonds, made by molecular evolution 1 1Edited by I. A. Wilson. Journal of Molecular Biology 275:245–253...sequences of hybridomas, but phage display is a popular way to generate scFvs.  For phage display, a pooled...oxidizing cytoplasm. expression in the presence of molecular chaperones that help scFvs fold properly in the...blocks for engineered proteins. This flexibility is largely due to scFvs being smaller than antibodies. scFvs...cells to a tumor site. ScFvs can also be fused to cellular toxins, radioisotopes, cytokines, and enzymes ...also used for eye diseases, such as age-related macular degeneration (AMD) since direction injection to...expressed on T cells to make CAR T cells. The extracellular scFv portion of the receptors recognizes an ...

... have a large impact on downstream results. Examples: “HTML”, “visual track”, “plot”, “tabular”, “interactive...Pasteur, Paris) that changed genome editing in the malaria community for the better (Nat. Biotechnol., 2014...Plasmodium falciparum (the parasite that causes malaria). I attribute their success to technical expertise...audience blindly receiving them; it results in highly similar content and the absence of iterative design; it...The third, WGE (published 15 Sep 2015), is more similar to EENdb than CrisprGE, but is also a good example...interactive”, and “save”. Trends: Most tools provide tabular HTML formatted output. Surprisingly only 2 tools... agnostic”, “model organisms”, “small genome”, “large genome”, and “organism biased scoring”. Trends: ...

...Pro tip! Getting a vector into a particular cell type depends largely on the viral capsid, but specificity...varying tissue tropism, allowing them to target particular cell types. Figure 1: Diagram of AAV ...cells. Generally, AAV serotypes have a primary cellular receptor and sometimes one or multiple coreceptors...contexts outside the nervous system to confirm its cellular receptors and tissue tropism. AAV5 AAV5 is the...additionally been reported to have tropism for liver, vascular endothelial, and smooth muscle cells (Issa et ...al., 2023; Wu, Asokan, et al., 2006). AAV6 is particularly well-suited for transduction of lung cells via... cross blood vessel barriers. It is especially popular for applications targeting liver tissue. In addition...

...on the intracellular side of the cell membrane. Heterotrimeric G proteins are important molecular switches...induce or inhibit intracellular signaling cascades as a response to the extracellular stimuli. Ligand dissociation...to the seven alpha helices that transverse the cellular membrane. The receptor recognizes the appropriate...signaling is initiated when a ligand binds to the extracellular surface of the GPCR. This results in a conformational...response in a certain cell type. Chemogenetics is similar to optogenetics but instead of light, these genetically...activate or inhibit neuronal firing. As GPCRs are the largest class of signal transducing receptors in the brain...

...targeting is led by a ~20 bp gRNA which is part of the larger CRISPR RNA (crRNA). In addition to the 20 bp guide...evolved ways to prevent degradation of important cellular RNA molecules with post translational modifications... Moving one position over in the sugar ring, a popular stabilizing modification is 3’ phosphorothioate...resulting in less off-target events. While both have similar advantages, bridged nucleic acids with N-methyl...et al., Frontiers in Genome Editing). This is particularly important in experiments in which viability ...transfection efficiency and even track localization in cellular experiments. There are many suitable dyes to choose...References and Resources References Hendel, A., Bak, R., Clark, J. et al. Chemically modified guide RNAs enhance...

... that actively engage with the community are particularly successful. Machine-learning denoising of raw...segmentation of imaging data (ILASTIK), or the popular plugin-based image process framework FIJI are just...such as the OpenFlexure microscope or open-source modular optical toolbox openUC2 are trying to overcome ...rely on such tools has allowed the community to regularly replicate designs worldwide. Distributing hardware...facilitates the reproduction of the structure in a similar way to the biological protocol, and assembly designs...Nevertheless, there are still many open questions to be clarified, especially with regard to licensing models. Direct...is a great example of how sharing of data and molecular tools can lead to faster and more efficient science...

...Mutagenesis is a tool that both evolution and molecular biologists use to tinker with DNA. Making changes...phenotypes, and forward genetics harnesses this at a large scale by screening diverse libraries of genetic ...must be grown in a turbidostat. Only mutate particular types of nucleotides. Both the Bassik Lab and...PolI3M that’s fused to a nicking Cas9 (nCas9). Similar to mutagenesis with error prone PCR, EvolvR uses...However, by using nCas9 to direct PolI3m to a particular genomic loci, EvolvR allows for targeted mutagenesis...replication by 120-fold over background. Due to its modular nature, the Dueber Lab created a few versions of...increasing the editing window of EvolvR to ~56 bp. A larger editing window also increased the likelihood that...

...key tools researchers use to prove causation in molecular biology and genetics. It can be used to characterize...are available for making small point mutations, large deletions/insertions require complicated, often ...possible to use the same technique just using a large primer known as a megaprimer. Piecing together multiple...methods can be expensive because they require particular enzymes, some of which are proprietary. These...have secondary structure or too much sequence similarity. REPLACR-mutagenesis: Letting bacteria do the...then joins the linear PCR products into stable circular plasmids - no nick repair is needed. Since the...L-arabinose. These cells can be made and frozen in large batches suitable for many experiments. When REPLACR-mutagenesis...

...another challenge associated with molecular plant parts, particularly for applied projects intended for...challenging to engineer a specific function in these multicellular organisms. However, the rewards are great if...biotechnology and provide a much needed boost to basic molecular plant research, more initiatives are needed to...associated with plant synthetic biology. Plant molecular biology has traditionally relied on overexpression...processes. Often the tools developed for plant molecular biology are specific to a given project, and like...pieces developed over the years were generated by large agricultural companies and carry associated intellectual...and the ocean. After receiving her bachelors in Molecular Biophysics and Biochemistry from the Illinois ...

... tumor’s backstage team. Microenvironments are cellular regions with specialized structures. Tumors are...neuroendocrine, adipose, immune, inflammatory, angiogenic vascular, endothelial, and stromal cells - a rich soup ...promoting proteins, such as human growth factor, vascular endothelial growth factor, and interleukin-6 (...2009). Then, scientists began focusing on the extracellular matrix and fibroblasts. With each cell mechanism...Centro Ciência Viva" in Aveiro, Portugal. She is particularly interested in science journalism and writing...BrbaraPinho6.      References Allinen, Minna, et al. "Molecular characterization of the tumor microenvironment...PMCID: PMC4788391. Kogure, Takayuki, et al. "Intercellular nanovesicle‐mediated microRNA transfer: A mechanism...

...Research Symposium organizer, David T. Riglar PhD. Here, Dr. Riglar discusses one of the six panel discussions...Many thanks to our guest blogger David T. Riglar! David T Riglar is a postdoc at Harvard Medical School.... problems that were identified last year.  In particular, at the symposium we will discuss where better...Jonathan Dinman, Chair of the Cell Biology and Molecular Genetics Department at University of Maryland ...collection about the scientific workforce, in particular about postdocs. Among other involvements, he ...science and public policy. Follow David on Twitter @driglar.     References 1. Alberts, Bruce, et al. "Rescuing...

...sign up for Google Scholar alerts. Based on keywords that you provide, Google Scholar will send you daily...Johnna's New Under The Sun Blog and is part of her larger tutorial series, A Beginner’s Guide to Writing ...useful to make literature searching and reading a regular part of your routine. Set a goal to read a certain...engines for finding scientific literature are Google Scholar and PubMed. You can search keywords, titles, authors...given research area. Unless you are looking for a particular protocol or are trying to replicate an experiment...possibilities until the results focus in on a particular conclusion. Many papers aren’t always written...of sufficient quality (error bars, statistics, clarity in images etc)? Did they perform all of the appropriate...