...Orientation)) is a powerful tool that provides precise temporal and spatial control of gene expression in vivo...- or reward-related behaviors. The high spatio-temporal precision and reversible modulation of FLEx vectors...

...the pSer-incorporated polypeptide. These three components were shown to effectively incorporate phosphoserine...Creating phosphoserine-incorporated proteins The generation of pSer-incorporated proteins requires a molecular...strain for pSer-incorporation The second component necessary for phosphoserine incorporation is a modified... actually been incorporated into the full-length protein. Up to 80% p-Ser incorporation into E17TAG GFP...different levels of incorporation due to differences in stability of the p-Ser incorporated protein as well...determine the stoichiometry of phospho-incorporated to non-phospho-incorporated proteins purified from the system...pSer-incorporated to non-phosphorylated protein. Verification of site-specific pSer incorporation can ...

...it is incorporated. We were surprised to see how rapidly the efficiency of mutation incorporation falls...contains the intended sequence changes to be incorporated into the edited genome. In mammalian cells such...these two strategies allows you to specifically incorporate homo- or heterozygous mutations. 1. Increasing...therefore strongly encourage any researcher to incorporate CRISPR/Cas-blocking mutations at both targeted... our frustration) in many cases we only saw incorporation of the blocking mutation. Despite the fact that... away from the cut site were not efficiently incorporated during HDR. We decided to systematically characterize...only about 30 bp, it is no longer feasible to incorporate mutations without screening thousands of clones...

...describing a technique that allowed for intentional incorporation of an unnatural amino acid in translation (Noren...natural translation machinery; these are often incorporated into the cells via plasmid. Third, a second ...containing the chosen codon that will result in the incorporation of the non-canonical amino acid at the site ...integrated into the cell’s genome, instead of incorporation via plasmid or media. Theoretically, if you ...will produce a protein with a site specifically incorporated ncAA, ready for you do to any number of experiments... (1989). A General Method for Site-specific Incorporation of Unnatural Amino Acids into Proteins. Science...Mehl, R., & Cooley, R. (2022). Site-specific Incorporation of Phosphoserine into Recombinant Proteins in...

...interacting proteins. No spatiotemporal control. Many PPI detection methods lack spatiotemporal control or an “...this limitation by creating Split-BioID, a spatiotemporally controllable version of the proximity-dependent...The Split-BioID method Enter Split-BioID, a spatiotemporally controllable version of the BioID protocol... other PPI methods, including BioID, is its spatiotemporal controlled labeling of interacting proteins...proteomics approach to monitor the composition of spatiotemporally defined protein complexes. Nature communications...

...self-assembly. In an effort to obtain even more precise temporal control of gene knockout or activation, the c-...)-FRB-NES). This system provides users greater temporal control over CRISPR/Cas9 mediated genome modification...activation allows scientists to spatially and temporally control which genes are turned on or off in a....0043072. Photoswitchable tools for spatial and temporal control of cell events Three years ago, Brian ...allows scientists to use light to spatially and temporally control endogenous gene activation. This system... anyone interested in understanding the spatio-temporal regulation of human genes. The plasmids are all...fluorescent reporters can be challenging enough, but incorporating multiple reporters into a single expression ...

...plasmid of interest. Scientists can also use electroporation, the application of an electrical charge to...lysogenic cycle, the phage’s genetic material is incorporated into the host’s genome at a particular integration... and stable cell lines, where foreign DNA is incorporated into the cell’s genome and is thus passed down...charged cell membrane. Physical methods such as electroporation or microinjection actually pokes holes in the...deliver nucleic acids to individual cells. Electroporation on the other hand uses electrical pulses to...

...azidohomoalanine (AHA). In the technique, AHA is incorporated into newly synthesized proteins instead of methionine...endogenous protein of interest that also had incorporated the AHA tag after pulse labeling of a determined...FUNCAT-PLA provides both quantitative and spatio-temporal information about the turnover of specific endogenous..."Puromycin inhibition of protein synthesis: incorporation of puromycin into peptide chains." Proceedings...Gabriel. "Inhibition by puromycin of amino acid incorporation into protein." Proceedings of the National Academy...

...want to turn things on or off, permanently or temporarily, we’ve got you covered. Defining your control...inducible? How long do you need control? Will temporary control (a few days) be enough, or do you want...than making a knockout and is a great way to temporarily reduce expression of essential proteins (when...expression (more on this below) will both achieve temporary repression of protein expression. Addgene has ...via a custom gRNA, to your gene of interest to temporarily increase expression. There are several flavors...

...delivery include plasmid transfection, RNP electroporation, and viral transduction; but these methods ...Gag protein fused to a Flag-tagged SpCas9 is incorporated into Nanoblades, just like the unmodified MLV...MLV Gag protein. After Gag-Cas9 is incorporated, the MLV protease, supplied as part of the MLV Gag-Pol...gene, while editing was not detected in cells electroporated with RNPs using the same guide. Nanoblades ...influence the level of off-target effects, with electroporation of RNPs generally having lower off-target edits...

...not millions, of plasmids, they are generally electroporated into electrocompetent cells to ensure the highest...chemical transformations is less than that of electroporation, but an advantage is that chemical transformations...protocol, the size of the cuvettes your lab’s electroporator uses, etc. The DNA amount can be anywhere from...DNA in 100 μL of cells over four individual electroporations. The specifics of this can be based on the...5,000,000 30 bp At least 200 ng Liquid Electroporation Protocol (DOCX, 15.8 KB) Barcode SPLINTR...

...and gRNA expression       Plasmid DNA may be incorporated into the genome     Off-target effects can ... common techniques for delivery of RNPs is electroporation (A in the figure above), which generates pores...through a process known as CRISPR-EZ (CRISPR RNP Electroporation of Zygotes) (Chen et al. 2016). For CRISPR ...CRISPR experiments that involve HDR, electroporation can be combined with cell-type specific reagents in a ...genome editing by CRISPR ribonucleoprotein electroporation of zygotes." Journal of Biological Chemistry...

...FLP-FRT. In addition, a variety of means to spatiotemporally control FLP and Cre expression have been developed... only occur at a specific time point? To allow temporal control, ligand regulated Cre and FLPe recombinase...control of Cre recombinase. After in-utero electroporation, this plasmid mix results in sparse labeling... brain area can be transfected by in utero electroporation. To achieve sparse labeling of individual neurons...instance) can be controlled with FLP-FRT. For further temporal control, Tamoxifen inducible Cre versions can ...

...of Brainbow, Brainbow 3.0 (Cai et al., 2013), incorporates the highly photostable fluorophores mOrange2...default, they are not fluorescent. Instead, they incorporate a truncated, non-fluorescent form of YFP as the...overcame the low expression of XFP of Brainbow by incorporating a Tet-Off system to boost XFP expression. The...Tetbow can be introduced in vivo either by electroporation of plasmids, or using AAV vectors. The Imai... lab also developed a version of Tetbow that incorporates chemical tags instead of XFP to optimize it ...

...the first fluorescence microscopy technique to incorporate optical sectioning. Optical sectioning refers...their fast movements. Depending on the sample, a temporal resolution on the millisecond scale may be needed...resolution due to the lack of a requirement for high temporal imaging speeds. Standard fluorescence widefield...sectioning, imaging over large distances and high temporal resolution. Traditionally, this type of sample...technique for many of these samples due to its fast temporal resolution and decreased phototoxicity. Lightsheet...

...you like the gene to be incorporated into the genome? Some vectors incorporate into the genome randomly... of interest in a plasmid and get the vector incorporated into the germline. To begin this process, you...pick out flies where your transgene has been incorporated into the germline. Outsourcing the plasmid injections...

... optogenetics is renowned for enabling precise temporal and spatial control. Optogenetics uses genetically... on and off gene expression while imparting spatiotemporal control, reversibility, and repeatability. ...codon, TAG. (To learn more about site-specific incorporation of amino acids using pyrrolysl tRNA synthetase...) A Singular System with Precise Dosing and Spatiotemporal Control of CRISPR‐Cas9. Angew Chem Int Ed 58...Rauen RR, Kahn SI, Tsang M, Deiters A (2020) Spatiotemporal Control of CRISPR/Cas9 Function in Cells and...

...expression, sometimes incorporating chemically inducible approaches to provide temporal control, this new ...can deliver these components together using electroporation. In these bulk cell experiments, Clift et al...no stress or damage upon TRIM-Away, and the electroporated cells re-adhered to the culture plate within...

...gene incorporated into their genome. While transgenic mice have the new gene construct incorporated randomly...embryonic stem cells. Non-homologous recombination incorporates this DNA randomly into the mouse genome. In ...homologous recombination. A targeting vector is electroporated into embryonic stem cells and this will insert...

...FPs) were derived from natural photoreceptors incorporating biliverdin IXα (BV) autocatalytically. Fortunately... a cell and depends on the efficiency of BV incorporation by an apoprotein and protein expression level...Oliinyk et al., 2019). They were engineered to incorporate BV. Their advantages are their small size (17...Michnick, S.W. An infrared reporter to detect spatiotemporal dynamics of protein-protein interactions. Nat...