...various mutations used to increase its stability activity, can be readily purified using plasmids found ...protein since the enzyme cleaves the SUMO tag in the same manner as it would have in the context of a cell...

... However, at the same time, it's fun to be involved in these professional activities because I can contribute...

...It was discovered as an antigen upregulated on activated lymphocytes and is commonly used to enrich adult...Deverman group then looked to see if PHP.B required the same receptors that its parental AAV9 relied on for transducing...

...target gene Many different activators exist, including the multi-plasmid SAM system. Select Expression ...application. For example: To activate or repress a target gene using dCas9-activators or dCas9-repressors, gRNAs...effective than dCas9 alone for mammalian cell lines. Activate (CRISPRa) Increase expression of an endogenous...without permanently modifying the genome dCas9-activator (such as dCas9-VP64) gRNA(s) targeting promoter...expression of CRISPR components Short window of CRISPR activity may decrease off-target effects Additional Resources...gRNAs based on predicted on-target and off-target activity A PAM sequence is absolutely necessary for Cas9...high-fidelity Cas enzyme. In addition to off-target activity , it is also important to consider factors that...

...dCas9, zinc finger proteins, and transcription activator-like effectors (TALEs), with fifteen total variants...protect fragile samples during cryo-EM grid plunge freezing By Rachel Leeson Cryo-EM samples can be damaged...and structure determination LEA proteins protect samples during cryo-EM grid freezing Engineered and evolved...antibodies for marking astrocytes MagIC-Cryo-EM for sample enrichment and structure determination By Mike ...enrichment streamlines the purification and minimizes sample loss while also supporting imaging, making it easy...cryo-EM grid.     Figure 1: MagIC-cryo-EM for sample enrichment and structure determination. A) Schematic...beads for scarce macromolecules in heterogeneous samples. bioRxiv 2024.01.21.576499; doi: https://doi.org...

...the integrin from its closed to its open state, activating the receptor. Ligands can bind to integrins in...Cells often express multiple integrins that bind the same ligand, making it difficult to dissect the function...-open. In the two closed states, integrins are inactive, with low affinity for ligands. When integrins...extracellular matrix. During immune responses, this same conformation encourages immune cell adhesion and...IPI antibodies  Antibody Target Proteins Reactivity Isotype Source Species Anti-Integrin alpha...

... design has been shown to increase T7 promoter activity. To avoid re-cutting by Cas9 after editing, make...They have successfully used guides targeting the same or complementary strands, as well as guides facing...

... as increased specificity and lower off-target activity (Hu et al., 2018).  In 2020, the Kleinstiver lab...and crRNA (or synthetic gRNA) are derived from the same species. Expanding the CRISPR toolbox The isolation...

...homology-directed genome editing by catalytically active and inactive CRISPR-Cas9 using asymmetric donor DNA. Richardson...by intracellular factors such as nucleases and reactive oxygen species, or external forces such as ionizing... occur; however, if both HJs are resolved in the same manner, this results in a non-crossover event.  ...

...effect within the body, or they can be used to activate cellular pathways by binding to cell surface receptors... exact amount of protein in your sample, you’ll compare your samples to a standard curve. This curve is...the known samples is used to calculate the concentration of protein in the experimental samples. Get more...multiple proteins on the same cell simultaneously as well as compare between samples. Learn more about flow...of different isotypes can be used together in the same experiment because the reagents used to detect the... population of antibodies that all recognize the same epitope of the target protein. Most commonly these...expression in cell lysates, whole cells, or tissue samples. Many experiments that use antibodies use a primary...

...expressing TH protein indicating that there is promoter activity. Additionally, some neurons express low levels... with the same titer and serotype previously used in a different brain region that the same result will...sites will be flanked by two lox sites now in the same orientation. Next, the loxp site is excised since...since it is between two lox2272 sites in the same orientation preventing the transgene from reverting. Figure...scrambled one pair of lox sites and either used the same sequence to flank a transgene (homologous pairs)...the DIO/FLEx vector. This step is essentially the same as what was used to optimize the serotype, except...

...chemogenetic tools, where modification of cellular activity outside of your experiment site or circuit can... different phenotypes of the same cell type, are intermingled in the same brain nucleus: a serotype that...tropism between animal species or even strains of the same species. What works well in a mouse may not work...doi.org/10.3390/cells12050785 Murlidharan, G., Samulski, R. J., & Asokan, A. (2014). Biology of adeno-...Response against the Tetracycline-Dependent Transactivator Correlates with Long-Term Doxycycline-Regulated...10.3390/pharmaceutics13050750 Murlidharan, G., Samulski, R. J., & Asokan, A. (2014). Biology of adeno-...

... hybrid vector, as measured by β-galactosidase activity, was 81% of that from a single vector, but significantly...nbt0702-697 Hirsch ML, Agbandje-McKenna M, Jude Samulski R (2010) Little Vector, Big Gene Transduction:...Bellon I, Yin C, Chavala S, Pryadkina M, Richard I, Samulski RJ (2013) Oversized AAV Transductifon Is Mediated...

...RNA cleavage, and translational repression or activation: Kellermann, Stefanie J., and Andrea Rentmeister...plasmids used to generate these fusions all use the same restriction enzymes for cloning. Furthermore, some...

...far-red genetically encoded indicators of neuronal activity. J Neurosci Methods 362, 109314 (2021). Yu, D...., and deep penetration of red-shifted light. The same NIR FPs and biosensors can be imaged at spatial ...

... metastasis as well as teaching and mentoring activities. Follow her on twitter @zcastan or on linkedln... my mentor, since she does research at UK in the same field that I do: prostate cancer. She has guided... Pilot Program 2015-2015 Scholars need strong proactive mentors who have relevant expertise and skills...

...Griesbeck, Heim, & Tsien, 1999), monitoring kinase activity (Ni, Titov, & Zhang, 2006), quantifying antibody... However, the acceptor doesn’t need to go to the same exact location as the donor. As long as some of ...targeted nearby (such as a neighboring house on the same street as the donor), then FRET measurements are...Ebright, 2001). These Cy/NTA conjugates work the same way as NTA-sepharose beads used for protein purification... binding sites are adjacent to each other on the same protein and a negative control experiment conducted...

...effects, each group finds very low to no off-target activity at the ten highest-ranking predicted off-target...slightly different methods, they each observed the same positive phenotypes in cardiac and skeletal muscle...

...protein level at the timepoint studied Off-target activities of the RNAi Correctly interpreting your data...the abundance of the epitope in the sample, relative to cross-reactive antigens – that is, an antibody shown... recognize different epitopes of the same protein, show the same pattern of labeling, this is supportive...detecting the target-of-interest in your assay and sample-of-interest. The approaches available broadly map...determining the selectivity of antibodies for other sample types and for non-surface (intracellular) antigens...approaches used will be different depending on the sample and target type. To start the validation process...the target at a robust level, similar to that in samples of interest. If you are working with cells that...

...Resources on the Addgene Blog Learn to Use Cas9 Activators Perform Genome-Wide Screens with CRISPR Generate...investigator can be relatively certain that their samples don’t have off-target mutations if they remain ...