We narrowed to 743 results for: UTY;
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TypeBlog Post... GCaMP containing plasmids are available for distribution. If your experiment involves delivering a calcium...types of searches including depositing labs and institutions. For example, search for “Melina Fan” and you...Lab published in eLife. Addgene is currently distributing 94 plasmids expressing validated antibodies ...
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Plasmids 101: Protein tags
TypeBlog Post... be to fuse an affinity tag to it. The FLAG, hemaglutinin antigen (HA), and c-myc tags have been the workhorses...KRRWKKNFIAVSAANRFKKISSSGAL 4 Affinity and Purification Binding and elution steps use very moderate buffer conditions FLAG...cytometry, but rarely used for purification as elution requires low pH poly His HHHHHH 0.8 Affinity... -
To Codon Optimize or Not: That is the Question
TypeBlog Post...in vivo experiments and in biotechnology and therapeutics (Mauro and Chappell 2014). Ultimately, it is...critical analysis of codon optimization in human therapeutics. Trends in Molecular Medicine 20:604–613 https...ZE, Kimchi-Sarfaty C (2011) Understanding the contribution of synonymous mutations to human disease. Nat... -
CRISPR-mediated Plant Base Editors
TypeBlog Post...by Gaudelli et al. resulted in the successful evolution of an adenosine deaminase that can act on DNA ...for surviving colonies and subsequent rounds of evolution and engineering gave rise to a mutant TadA (TadA...in rice. In Genome Engineering: The CRISPR-Cas Revolution, Cold Spring Harbor Laboratory (2018). Additional... -
New CRISPR Tools: Cas7-11 and PASTE
TypeBlog Post...The hardest part of any revolutionary discovery, including CRISPR, is turning potential into impact. ...collaborators of Omar and Jonathan and experts in CRISPR evolution, were also working on the project. Using computational...possibilities at the bench and potentially in therapeutic applications. The potential of CRISPR has yet... -
Viral Production at Addgene
TypeBlog Post...Briefly, 293T cells are transduced with serial dilutions of a lentiviral vector, incubated for 72 hours...line was obtained directly from Takara, and is routinely tested for mycoplasma contamination using mycoplasma...Cells transduced with our lentiviral vectors are routinely checked for signs of microbial contamination. ... -
Plasmids 101: Broad Host Range Plasmids
TypeBlog Post... Variation in ori structure allows plasmids to utilize diverse host replication machinery. For instance...Paracoccous denitrificans, Pseudomonas fluorescens, P. putida, Rhizobium meliloti, R. leguminosarum by. viciae... P. C., & Wong, H. L. (2023). Development of constitutive and IPTG-inducible integron promoter-based expression... -
Viral Vectors 101: Pseudotyping
TypeBlog Post...specific purposes as described previously (Table 1, Gutierrez-Guerrero, et al, 2020). Because retrovirus is ...disadvantages: Table 1 (Joglekar et al., 2017) Table 1 (Gutierrez-Guerrero et al., 2020) Table 1 (Cronin et al.,...–222 . https://doi.org/10.1006/mthe.2000.0116 Gutierrez-Guerrero A, Cosset F-L, Verhoeyen E (2020) Lentiviral... -
Plasmids 101: Codon usage bias
TypeBlog Post...; multiple codons encode single amino acids. Evolutionary constraints have molded which codons are used...express well in any organism. So what are the evolutionary constraints that lead to these preferences and...6. Pechmann, Sebastian, and Judith Frydman. "Evolutionary conservation of codon optimality reveals hidden... -
Using Video to Share Your Science: We Share Science
TypeBlog Post... variety of fields in order to provide useful solutions to complex challenges. As a cross-disciplinary...citation rates to measure the impact of their contribution. It offers a 5-step process and several examples...Social Networking for Scientists Read about the Evolution of Scientific Impact Data Freedom: The Expansion... -
Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus
TypeBlog Post...be dominant-negative and/or toxic. A potential solution is to include an in-frame degron, or degradation...in a single step and serve as donors for two consecutive CRISPR-mediated HDR events which results in the...Gibbs D, Kim SR, Maguire A, Rex TS, Di Vicino U, Cutillo L, Sparrow JR, Williams DS, Bennett J, Auricchio... -
Harnessing Bacterial Toxins for Allelic Exchange
TypeBlog Post...resistance marker from the vector (abR, Figure 1). The resolution step (“double crossover”) which leads to the ... program) (Latynski and Valentovich 2014). We routinely clone the purified amplicons upstream and downstream... in 5 days. Counter-selection with sacB has revolutionized allelic exchange. Even now, only half our lab... -
The Fluorescent Vegetables in Aptamer Soup
TypeBlog Post...fluorescent aptamer, was identified through directed evolution. First, a pool of RNAs that activate the fluorescence...fluorescence of DFHBI were identified using systematic evolution of ligands by exponential enrichment (SELEX). ... by fluorescence-based selection and directed evolution." Journal of the American Chemical Society 136.46... -
New Optogenetic Tools for Cytoskeleton and Membrane Control
TypeBlog Post...activity of the original, generated using directed evolution (Tei et al., 2023). PLD can hydrolyze phosphatidylcholine...matter. Plus, for folks interested in directed-evolution methods, the activity-based selection in mammalian...Baskin, J. M. (2023). Activity-based directed evolution of a membrane editor in mammalian cells. Nature... -
CRISPR 101: Homology Directed Repair
TypeBlog Post...external forces such as ionizing radiation, chemotherapeutics, and ultraviolet light. In this post, we will...via cleavage in one of two ways - each junction resolution could happen on the crossing strand (horizontally...repair and genome engineering CRISPR-Cas9 has revolutionized the genome engineering world and made targeted... -
Site Directed Mutagenesis by PCR
TypeBlog Post...can be used to introduce specific nucleotide substitutions (or deletions) in a tailored manner. The approach...perform alanine scanning mutagenesis or targeted substitution of key residues), and in SNP analysis (to introduce... bases at the 3’-end interfere with plasmid reconstitution, and consequently Taq polymerases cannot be... -
Hot Plasmids: Spring 2025
TypeBlog Post...much to learn about their myriad functions and therapeutic potential. Our partners at the Institute for ...proteins. Lucky for us, we now have a two-for-one solution: TurboCas!TurboCas provides the genomic targeting...-type-specific expression has been a promising solution. Recently, the labs of Gord Fishell and colleagues... -
Plasmids 101: Dimers and Multimers
TypeBlog Post...and accessibility, more researchers have been utilizing full plasmid sequencing services to verify the...independently increase the frequency of base substitution in human cells. J Biol Chem. 2011 Mar 25;286...E, Fournes F, Cornet F, Hallet B, Rousseau P. Resolution of Multimeric Forms of Circular Plasmids and ... -
PiggyBac-ing Through the Genome Editing Field
TypeBlog Post... transcription factors, or when the gene is constitutively-active or a dominant-negative mutant. Reducing...piggyBac™ transposase. This can be best achieved by utilizing a piggyBac™ transposon in a donor plasmid carrying...and M. H. Wilson, “piggyBac-ing models and new therapeutic strategies.,” Trends Biotechnol., vol. 33, no...Resources on the Addgene Blog Learn about the Sleeping Beauty Transposase Read up on Genome Engineering with ... -
A Guide to Getting Started in Undergrad Research
TypeBlog Post...involved in the science, experimental design, and execution. Can lead to an independent project. Lab technician...Other roles you might see Lab manager: managerial duties ex: order materials, lab maintenance Administrative...Administrative assistant: assists PI with admin duties Staff scientist: similar role to a postdoc but more permanent...