We narrowed to 330 results for: iCre

...the region between the microhomology and the retention of a single microhomology sequence. For more mechanistic...Step 1: Generate microhomologies in the PITCh vector ~20 bp 5’ and 3’ microhomologies are added to the...double-stranded break repair mechanism: MMEJ (microhomology-mediated end-joining)? MMEJ, a form of alternative...contrast, MMEJ uses regions with 5-25 bp of microhomology flanking a DSB to repair DNA. The DNA ends are...precise deletion of the sequence between the microhomologies.MMEJ has certain advantages over HDR. Some species...vector should be constructed with 5’ and 3’ microhomology to the insertion locus flanking the GFP-Puro... cassette and one in between the 5’ and 3’ microhomologies in the genomic locus. The first two breaks ...

...specific targeting of microglia with AAV By Alyssa Neuhaus Genetic modification of microglial immune cells is...valuable tool for microglia research and broader therapeutic applications.  Find microglia-targeting AAV ...Voltron2 voltage sensors AAV vectors for targeting microglia The REVeRT Dual AAV Vector System Neurodegeneration...for central nervous system research; however, microglia are notoriously resistant to viral transduction...IBA1 promoter (hIBA1a, 466 bp) that drives high microglia-specific gene expression when incorporated in ...Efficiency and specificity of these AAV vectors for microglia was further enhanced by packaging the virus as...transgene expression in neurons and other non-microglia cells.      Figure 5: A) Representative...

...the Addgene repository in 2015 was... lentiCRISPR V2   LentiCRISPR v2 originates from Feng Zhang’s lab... and how does it work? LentiCRISPR v2 purpose As its name suggests, lentiCRISPR v2 is a lentiviral transfer...,000 human and mouse genes respectively, use lentiCRISPR v2 as the backbone for their many gRNAs. The ...high titer lentivirus production afforded by lentiCRISPR v2 is key to its efficient use in screening experiments...pooled library screens. Availability and Use LentiCRISPR v2 is available from Addgene both as a single...CRISPR/Cas9. If you need help cloning a gRNA into lentiCRISPR v2, you can take a look at the protocol kindly...kindly provided by the Zhang lab. Even though lentiCRISPR v2 has only been available for a little over a ...

...PubMed Central PMCID: PMC3984138. Microscopy U. Basics of FRET Microscopy. Additional Resources on the Addgene...Visualization can be achieved using fluorescent microscopy - even in living cells and whole organisms. You... many Fluorescent Proteins (FP) available for microscopy, by cloning your protein of interest into a vector...must bear in mind that the resolution of light microscopy is limited by the wavelength of light and, practically...practically speaking, for common lab microscopes the detection limit is 200 nm. The size of most proteins...10 nm and therefore colocalization detected in microscopy cannot be interpreted as direct molecular interaction...without further investigation. One fluorescent microscopy method commonly employed for analyzing interactions...

...technology can have on the human microbiome. You can consider the microbiome as a collection of organisms,... honey bee and bumble bee gut microbiomes titling it the Bee microbiome toolkit (Leonard et al., 2018)... Pseudomonas syringae and Klebsiella oxytoca. Microbiome engineering Scientists have begun to realize ...methods of introduction. Engineering the bee gut microbiome Beyond the human gut, the Barrick lab has deposited...plasmids that can be used to engineer the bee gut microbiome. They created a modular system using a broad ...-Cas nucleases to produce sequence-specific antimicrobials. Nat Biotechnol 32:1146–1150 . https://doi....RJ, Mimee M, Lu TK (2014) Sequence-specific antimicrobials using efficiently delivered RNA-guided nucleases...

...experienced this at the ASM General Meeting (now Microbe) in 2013 in Denver, CO. While I had been to the...scientists and some rising stars in the world of microbiology, some before they had even established their...Porch with old and new friends while attending ASM Microbe in Atlanta, GA, in 2018. A lot of my experiences...from participants in Binning Singletons at ASM Microbe in 2019 were overwhelmingly positive. Susanna Harris...remarked that “This was not my first time at the ASM Microbe meeting, but it was my favorite because I finally...better Our experience running this program at ASM Microbe in 2019 is written up in mSphere. A post-meeting...future to match people within similar fields (ASM Microbe has eight tracks), something we didn’t have enough...

...imaged in distinct fluorescent channels on the microscope. With the proliferation of fluorescent proteins...and that can be imaged with the optics on the microscope(s) you intend to use. An accurate determination...your fluorescent proteins compatible with your microscope optics? To determine if the fluorescent proteins...you are interested in are compatible with your microscope optics, you will want to compare the excitation...protein with the filter sets or lasers on your microscope. Ideally, you would like to have substantial ..., so that the protein is well excited by your microscope and the fluorescence emission of the protein ...protein is efficiently collected by the microscope. To compare the match between a fluorescent protein and a...

...with BioRender.com. Resistance (R) plasmids: microbial defense Genes allowing bacteria to survive toxins...resistance to them are the tools of a millennia-old microbial war that humans only noticed last century! Nevertheless...implications for public health. Bacteriocin plasmids: microbial offense One way to triumph over your neighbors...plasmids” or “Col factors.” Virulence plasmids: microbial infection Virulence plasmids encode genes that... transform plant cells. Degradative plasmids: microbial hardiness There are also less aggressive ways ...of metabolic pathways. Fertility (F) plasmids: microbial sharing F plasmids contain the genes for bacteria...through a population of cells. Cryptic plasmids: microbial mysteries Cryptid hunters watch out — cryptic ...

...home edition, the MicrobiGals created glow in the dark petri dish ornaments and microbe cookies! 🤗 🤗 ...Td2M6AOpuW pic.twitter.com/ofLI4eD8Zs — Microbigals (@MicrobiGals) December 23, 2020   In second place,...bacteria, fungi, viruses, and one microbe ornament even dyed with real microbial pigment. Check out the hashtag...place, Dr. Anne Madden created a microbe advent tree displaying over 25 archaea, bacteria, fungi, and viruses...viruses. Final tree with the complete #MicrobeAdventCalendar ornament set (plus more!) It features archaea...

...long dsRNA, typically hairpin RNA, or artificial microRNA). Using a DNA template for the RNAi trigger is... hpRNAs through Golden Gate cloning. Artificial micro RNA and short hairpin RNA The greatest limitations...shortcomings, both short hpRNAs (shRNAs) and artificial micro RNAs (amiRNAs) are frequently used. Although similar... are engineered from endogenous precursors of microRNAs found in the genome, whereas shRNAs are exogenously...hairpin RNA (shRNA) is conceptually similar to the microRNA precursor mentioned above and is a popular RNAi...limited, single cell protoplast analysis. Instead, microscopic plants must be visually identified as undergoing...the entire culture can be harvested instead of microscopic isolation. The plasmids required to conduct an...

... your plans. This situation may arise for a microbiologist, where cloning two plasmids into one bacterium...deal with the more sinister plasmids of the microbiological world. (Want to know more about these plasmid...beginning to use plasmid incompatibility to combat antimicrobial resistance. Using novel plasmids based partly... and Control of Circular Bacterial Plasmids. Microbiology and Molecular Biology Reviews 62(2): 434–464...assortment of pure plasmid clusters. Molecular Microbiology 56:1430–1440 . https://doi.org/10.1111/j.1365... plasmid incompatibility. Letters in Applied Microbiology 47:235–240 . https://doi.org/10.1111/j.1472-...19241516 Novick RP (1987) Plasmid incompatibility. Microbiology Reviews. 1987;51(4):381–395. Pubmed PMID: 3325793...

...- a chemically stable tool for microscopy by: Susanna Stroik Microscopy techniques have been moving at...advanced microscopy applications such as super resolution, electron, and expansion microscopy. hfYFP is...hfYFP in expansion microscopy as well as correlative light and electron microscopy and observed high fluorescence...re looking for a super stable FP for your next microscopy experiment, consider hfYFP and LSSmGFP.   ...Chemically stable fluorescent proteins for advanced microscopy. Nat Methods 19, 1612–1621 (2022). https://doi.org...

...contributed by guest blogger, Jessica Sacher, a microbiology PhD student at the University of Alberta studying..., see Simpson et al., 2015). Visit Addgene's MIcrobiology Pages The problem: Host binding proteins are... blogger Jessica Sacher! Jessica Sacher is a microbiology PhD student at the University of Alberta studying...new applications for old resources." Trends in microbiology 23.4 (2015): 185-191. PubMed PMID: 25708933....bacteriophage-based therapeutics."Frontiers in Microbiology 7 (2016). PubMed PMID: 27536293. PubMed Central...metagenomic screening." Applied and environmental microbiology 74.5 (2008): 1649-1652. PubMed PMID: 18083885...Learn How CRISPR Can Be Combined with Phage for Antimicrobial Purposes Use Yeast Two Hybrid Systems to Probe...

...using i-GONAD (15%) vs. microinjection delivery (25-67%). However, microinjection does require 2.5x more...for clones with the recombined product, you’d microinject into blastocysts to be implanted in pseudopregnant...efficiency. Easi-CRISPR is compatible with standard microinjection and electroporation methods using in mouse ...and delivery methods Using RNP delivery and microinjection, the efficiency of Easi-CRISPR for simple knock-ins...anatomy with the equipment used for i-GONAD. A micropipette is used to inject solution into the oviduct ...components to the germline. In contrast to standard microinjection, i-GONAD is performed directly on pregnant ...electroporation. Ohtsuka et al. compared traditional microinjection and i-GONAD knock-in efficiency using a small...

..., check out the OpenWorm website. microPublication microPublication is a unique publication for the worm...otherwise be lost in lab notebooks forever. microPublications are peer-reviewed and assigned a DOI that ...referenced. In addition the data published in microPublications is curated and added to other databases including...including wormbase and flybase. microPublication is run and maintained by members of the worm community... four-dimensional contrast interference (DIC) microscopy and computer imaging processing. The combination...introducing point mutations that was published in a microPublication (Schwartz and Jorgensen, 2018).  These are...point mutations with unc- 119 + selection." MicroPublication 1 (2018): 2–3. Wang, Han, et al. "cGAL, a ...

...and electron microscopy (CLEM) is an imaging technique that combines electron microscopy (EM) with fluorescence...the second, and vice versa. Finally, as every microscope will have it’s own quirks (level of zoom, intensity...to revolutionize the field of super resolution microscopy. PA-FPs, especially the monomeric, reversible... Indeed, within the world of super resolution microscopy, PA-FPs have helped create several new imaging...methods, including photoactivation-localization microscopy (PALM, also FPALM) (11). For PALM, a small area...engineered. By combining PA-FPS with superresolution microscopy, scientists now have at their fingertips the ...mCherry for high-resolution two-color fluorescence microscopy." Nature methods 6.2 (2009): 153-159. PubMed ...

...Strategic immune evasion mechanisms - The tumor microenvironment Through interactions with the immune system...and thrive within what is known as the tumor microenvironment - a vital atmosphere created by the cancer...to mature into effector cells in the tumor microenvironment. Tumor cells in this environment produce growth...recruit regulatory T cells (T-regs) into their microenvironment by producing C-C motif chemokine 22 (CCL22...function. Creating Tolerance T-regs in the tumor microenvironment produce CTL associated protein 4 (CTLA-4),... above. Finally, signals released from the microenvironment help create tolerance beyond direct proximity...growth and subjecting patients to the risk of microbial infection. Chronic inflammation from infection...

...whistles’ – including microscale light emitting diodes, thermal sensors, and microfluidic channels. These features...neural fibers developed by the Anikeeva lab are microelectronic fibers made from thermoplastic polymers that...desired thickness. The fibers are drawn with microelectronics included in them for downstream control – ...(Orguc et al., 2020). Using a programmable microcontroller, NeuroStack coordinates optical stimulation...A., … Anikeeva, P. (2023). Multifunctional microelectronic fibers enable wireless modulation of gut and...resources on the Addgene blog Plasmid tools for microbiome studies Channel rhodopsins for optogenetics Viral...

...sterilization as mycoplasma can even escape 0.2-micron filters. Figure 1: Contamination! Tissue culture...media that turns yellow from lowered pH due to microbial contamination. Photo by Kaustubh Kishor Jadhav...Detection of mycoplasma by naked eyes or optical microscope can be very difficult, so how do you know it’...mycoplasma. When observed under a fluorescent microscope, the mycoplasma DNA appears in the form of small...lines for assay or analytical purposes. Use 0.1-micron filters as are they are a better way of avoiding...avoiding mycoplasma, rather than using 0.2-micron filters, for media filtration. There is no 100% assurance...the Detection of Mycoplasmas Using Fluorescence Microscopy. Cells 8:1510 . https://doi.org/10.3390/cells8121510...

...Fluorescence Microscopy. iBiology. https://www.ibiology.org/talks/fluorescence-microscopy/   Uhlen M, ...Learn more about fluorescence microscopy in Which Fluorescence Microscopy Techniques is Best for Me? Read...commonly proteins. You can then use fluorescent microscopy to look for those fluorescent molecules, knowing...fluorescence, try the Introduction to Fluorescence Microscopy video from iBiology). IHC and ICC are essentially.... The methods used depend on the sample type, microscopy tools available, and your experimental goals....