We narrowed to 321 results for: POR C

...into an attL-containing entry vector.   Method C: Restriction cloning of a restriction enzyme fragment...attL-containing entry vector.   Figure 5: Method C to create an entry clone: Restriction cloning the ...coding sequence. If you choose this strategy, it’s important to include the proper protein expression elements...Clone When making the expression clone, it is important to choose the destination vector that best fits...

...create an N- or C-terminal fusion based on your experimental needs. Equally important, before cloning ... retrieval sequence only functions at the extreme C-terminus [3]. You must therefore determine your cloning...potentially affect function [5]. We and others have reported that FP cysteine residues form inappropriate disulfide...

...fluorescent proteins (GFP and YFP) and protein tags (c-MYC and HA). We have also created the Monoclonal ...repositories, such as Addgene and the DSHB, play an important role in scientific exchange. In addition to ensuring...provide standardized quality-control and technical support to the community. Thanks to Our Guest Blogger! ...

... a AAV8 expressing eGFP under the human ubiquitin C promoter. Anatomical abbreviations: Gr= Gracile tract...Deisseroth. (2014) Nat Protoc., 9, 1682-1697. Pubmed. C. Soderblom, D.-H. Lee, A. Dawood, M. Carballosa, et...addition, the use of viruses allows much better spatiotemporal control over labeling methods as compared to...

...similar nuclease folds found in other proteins. The C-terminal domain (CTD) of the protein also belongs ...https://doi.org/10.1038/s41586-022-04470-1 Cofsky, J. C., Soczek, K. M., Knott, G. J., Nogales, E., & Doudna...Sternberg, S. H., Redding, S., Jinek, M., Greene, E. C., & Doudna, J. A. (2014). DNA interrogation by the...it is limited to only 20 bases, or why it’s so important to choose a target sequence with a nearby protospacer-adjacent... lobes laying open like a book (Figure 2). The portion of the CTD that recognizes the PAM is so flexible..., but they improve cutting efficiency and are important for robust activity in cells (Jiang & Doudna, ... guide RNA stabilizing the DNA-binding protein portion of the CTD, ready to check for PAM sequencines....

...References and resources Milling, L. E., Markson, S. C., Tjokrosurjo, Q., Derosia, N. M., Streeter, I. S....screens have been used to interrogate T cells, an important immune cell type, in more complex and physiological...

...reactions, where a fraction of one of the four bases (A, C, T, or G) is supplied as a dideoxyribonucleoside triphosphate...regions of interest, or complex sequences such as G-C rich areas, secondary structures, or repetitive regions... technology, but Oxford Nanopore is one of the most popular options. Nanopore sequencing deviates from...pulled through the pore via motor proteins. When each nucleotide passes through the nanopore, the electrical...3: Overview of Nanopore sequencing technology. Created with BioRender.com. Nanopore can provide longer...sequencing methods (Figure 3). It uses (aptly named) “nanopores” that are attached to the flow cell membrane and...-read sequencing. At Addgene, we routinely use Nanopore technology to sequence our packaged viral DNA ...

...these methods a try.  References:  Branon, Tess C., et al. "Efficient proximity labeling in living cells...1837. PubMed PMID: 30676735. Kim, D. I., Jensen, S. C., Noble, K. A., KC, B., Roux, K. H., Motamedchaboki...the mitochondrial calcium uniporter (Martell et al., 2012). It is important to note that the biotin-phenol... of exogenous biotin allowing for more precise temporal control.  Find plasmids for TurboID and miniturbo... where two proteins are fused to complementary reporter proteins that can only assemble when in close ... ascorbate peroxidase as a genetically encoded reporter for electron microscopy." Nature biotechnology...proteomics approach to monitor the composition of spatiotemporally defined protein complexes." Nature communications...

...specificity of GFP expression for the indicated ssAAV5s. C) Quantifications showing high microglia transduction... microglia-targeting AAV plasmids here! Serrano, C., et al. (2023). Simple and Highly Specific Targeting...2024). They developed a new prime editor that incorporates La’s N-terminal domain, PE7 (Figure 2).  PE7...high microglia-specific gene expression when incorporated in an AAV vector (Figure 5). Efficiency and ...including in human retinal organoids and live mice. Importantly, REVeRT avoids the introduction of extraneous...associated with viral vector-based therapies. REVeRT supports advanced gene expression regulation, enhancing...

...due to poor folding and thermal instability at 37°C (Ouellet et al., 2016) and requires a tRNA scaffold...imaging single molecules and for live-cell imaging in C. elegans. Other groups have used Mango to study RNA-protein...split aptamer is composed of a “Top” and “Bottom” portion, each fused to a separate piece of RNA. Fluorescence...photostable, and were presented in the study to support the uniqueness of the photostability of the Corn-DFHO...growth as RNA biology, we are looking forward to reports on the next additions to aptamer soup! References...

...frozen glycerol stocks. It is very important that these stay frozen at -80 °C, since multiple rounds of freezing...; CDSns, coding sequence with no stop codon; TAG, C-terminal tag; TER, terminator. Created with BioRender.com... perfect Our first tip, and arguably the most important, is to take some time to sit down, plan your cloning...

...into single-use aliquots, and stored at -80°C. Some reports suggest rapidly freezing virus in a dry ice... for less than a day, lentivirus can be kept at 4°C. For long-term storage, viral preps should be divided...protein attaches to cell surface receptors, other important binding events take place during the course of...

...conditions. Many recombinant proteins express better at 30°C or room-temperature, which is accomplished by growing...growing your culture to the desired density at 37°C and reducing the temperature or moving it to a bench-top...added to the growth medium. Induction timing is important, as you typically want to make sure your cells... optimal because of the high cell-density they support; however, minimal media supplemented with M9 salts...

...entire gene? Perhaps you are more interested in the c-terminus and would like to target that region. Or ... can be cloned directionally into the BbsI site. (c) The pCAG-EGxxFP target plasmid was co-transfected...expressing Cas9 and single guided RNA." Scientific reports 3 (2013). PubMed PMID: 24284873. PubMed Central...

...Zhi Ying Chen, Robert C. Robbins, Mark A. Kay, Michael T. Longaker, and Joseph C. Wu. "A nonviral minicircle...a Sendai Virus-Based Vector Using a Nonstructural C Protein and Endogenous MicroRNAs." Plos One 11, no...of Different Methods for Generating iPSCs. One important note: in this table, reprogramming efficiencies...high is based on the reprogramming efficiencies reported in González et al. A percentage is not listed ... be used to package reprogramming factors. An important milestone of reprogramming is the silencing of...cells for the presence of transgenes is still an important control. Additionally, Sendai virus doesn’t rely...comprised of OCT4, SOX2, LIN28, NANOG, and a GFP reporter. Minicircles are removed from cells by dilution...

...’s most common diseases. References Zhang Y, Yin C, Zhang T, Li F, Yang W, Kaminski R, Fagan PR, Putatunda...Gendron P, Zhu W, Guo F, Cen S, Wainberg MA, Liang C. CRISPR/Cas9-Derived Mutations Both Inhibit HIV-1 ...infection, including both common infections and opportunistic infections not usually seen in healthy individuals... genome had been excised, leaving only a small portion of the LTRs joined together. Clonal Cas9/gRNA-expressing...populations were also immune to HIV-1 reinfection. Importantly, no off-target effects were detected. Kaminski...patients. Although this is an exciting finding, it’s important to note that these HIV-1 patients were ART-naive...Cas9. While these results are discouraging, it’s important to note that these experiments were done in cell...

...Sato T, van Es JH, van den Born M, Kroon-Veenboer C, Barker N, Klein AM, van Rheenen J, Simons BD, Clevers...melanogaster. Hadjieconomou D, Rotkopf S, Alexandre C, Bell DM, Dickson BJ, Salecker I. Nature Methods. ...the 1980s, is one of the most important ways to spatially and temporally control gene expression, especially...ten colors (see table to the right); in various reports, Sanes and Lichtman have observed from 90-160 distinct...undergone recombination. This default state was disproportionately expressed by a majority of neurons, limiting... 8 colors. The use of AAV provides spatial and temporal control without the need for germline modification...refinements of the Brainbow technique will provide important insights into the complicated physical organization...

...expressed GFP in exogenous organisms (E. coli, and later C. elegans) for the first time. After Chalfie’s work...responsible for encoding GFP, and, in 1992, he reported its sequence. Soon after that, in 1994, Prasher...he lived! His hunch paid off, and he was able to report new fluorophores like DsRed from Discosoma sp. ...provided by the studies in Tsien lab, covered a portion of the color spectrum that GFP and its derivatives...reach. Nobel Prize for fluorescent proteins The importance of fluorescent proteins was recognized in 2008...

...involve a huge group getting from point A to B to C to D, etc. 7. Don’t let a tiny [or non-existent] ...that company culture and enjoying your job were important parts of team buidling - they relieved some of...

...solubility (Monnier et al., 2013). The linker keeps the C-terminus of one variable domain and the N-terminus...10.3390/antib2020193  Khantasup K, Chantima W, Sangma C, Poomputsa K, Dharakul T (2015) Design and Generation...cells to make CAR T cells. The extracellular scFv portion of the receptors recognizes an antigen of interest...interest while the intracellular T cell receptor portion of the CAR facilitates signal transduction and release...