...of human codon-optimized  cryptochrome 2 (CRY2) from Arabidopsis thaliana. When exposed to blue light, ...protein. After fusing the CRY2 and CIB1 domains to separate proteins of interest, a researcher can cause them...

...dimerization-dependent protein interactions. FEBS letters, 591 2, 415-424. PubMed PMID: 28032891. This is a different...feature is particularly useful for in-depth characterization of dynamic protein complexes. Split-BioID,...of the bait proteins’ interactome. Further characterization of these candidates is critical to understanding...

... already have an end product in mind, steps 1 and 2 will generally be straightforward. Likewise for step...present in a single lentiviral transfer vector or separate transfer vectors. May contain reporter gene (e.g...model.  If your expression system is not well characterized in terms of CRISPR use, you will want to invest...

...GUS-staining is a very useful visual reporter (Figure 2). The GUS enzyme cleaves a substrate, which will then...vincristine, are regulated in the medicinal plant, the Catharanthus roseus (Figure 1). The ultimate goal of our ...   C. roseus is not a model organism such as Arabidopsis or tobacco. Studying gene function in C. roseus...method for overexpressing genes In plants like Arabidopsis and tobacco, Agrobacterium infiltration into ...expression assay by agroinfiltration of leaves." Arabidopsis Protocols. Humana Press, 2006. 225-229. PubMed...transient gene expression analysis in seedlings of Arabidopsis and other plant species." Plant methods 5.1 (...expression method for analyzing gene function in Catharanthus roseus seedlings." Frontiers in plant science...

...system (by measuring the fluorescence intensity); and 2) to visualize the localization of the FP (fused to...experimental parameters (e.g. excitation light intensity, pH, or temperature). pH Stability: This parameter is ...Photostability can be affected by experimental parameters (e.g. excitation light intensity, pH or temperature...

...the necessary steps to perform:      Fig. 2: Quick-stepping to a FASTA file   With our outline...QC'ing data at Addgene is like standing under a Niagara Falls sized rainbow   Over the years we have...pipelines, including bioinformatics. We loved the parallels between Seqera’s mission and our own, as both ...the sequential tasks in our pipeline, we wrote a separate Nextflow process. In Nextflow, a process defines...concept of channels and implemented the input declaration correctly, the script block in the second process...

...inside the cell,” says Pedre Pérez. Figure 2: Micrographs of the biosensor expressed in the human...colleagues Filip Tomaska and Tim Hanson, are characterizing variants of iGluSnFR3 in vivo and have used...

...overview of these new variants:   Figure 2: Inner mitochondrial membranes imaged with structured...and screening that led to identification and characterization of AAV.CAP-Mac. Image reproduced from Chuapoco... Endothelial-specific AAV delivery By Cythina Arokiaraj (Gradinaru lab) Another useful viral tool deposited...

...You’ve probably heard that only 2% of our genome is made of protein-coding genes, and you might be wondering...transcriptome engineering References Jinek M, Chylinski K, Fonfara I, Hauer M, Doudna JA, Charpentier E. A Programmable...CRISPR-Based Approaches for the High-Throughput Characterization of Long Non-Coding RNAs. Non-Coding RNA. 2021...

...) 12 (>100) xCas9(3.7)-ABE 69 (1.4) 43 (2) 21 (3) Not examined 16 (>100) Average percent...xCas9 3.6 displayed the second best PAM range. Characterizing xCas9 in mammalian cells Hu et al. compared...SpCas9. Hu et al. performed GUIDE-seq analysis to characterize genome-wide off-targets, since increasing PAM...

... Werner S, Marillonnet S. PLoS One. 2011 Feb 18;6(2):e16765. doi: 10.1371/journal.pone.0016765. PubMed...few new ones that are not used so much but have comparable expression levels to things people do use a lot... developed mostly with model plants in mind, Arabidopsis and Nicotiana benthamiana. N. benthamiana is ...

...strain (which will be covered in more detail in part 2 of this blog series) controls the cell types in which...derived from random crosses between mice of uncharacterized backgrounds to create a limited but genetically...

...company culture.  It’s also a good idea to prepare 1-2 unique questions for each interviewer by looking at...of the theme of my interview stories as the characteristic which the story demonstrates I possess.   Bullet...

...of these issues are not particularly new. Chapter 2, “The Professional Scientist,” addresses the challenges...over two decades, attempts to answer it in five paragraphs, with examples. If that sounds ambitious, that...them wanting to learn more. There are individual paragraphs that may have warranted their own chapters, and...

...mazF gene and a gene for Zeocin resistance (Figure 2). This MazF-ZeoR cassette is flanked by two direct...or diluted as the cell grows resulting in the appearance of unmethylated sites in the chromosome. The ... without plasmid addiction systems. For example, Cara Morin and James Kaper used the hok/sok system to...223.1 (1992): 41-54. PubMed PMID: 1370544. Morin, Cara E., and James B. Kaper. "Use of stabilized luciferase-expressing...

...al (Nature. 1988 Jul 28;334(6180):320-5.) PubMed. 2. A segment of the 5' nontranslated region of encephalomyocarditis...multicistronic vectors simultaneously express two or more separate proteins from the same mRNA. We've discussed promoters...at the C-terminus of a 2A element, leading to separation between the end of the 2A sequence and the next...

...that numbers are not switched around or misread.  2. Track the container - It is just as important to ...to fulfill orders.  The barcoded tubes help to guarantee that no samples get lost during their journey....Addgene Get help with plasmid transformation, preparation, and analysis from our protocols and molecular...

...Hoogsten base pairs and wobble pairs.  Figure 2: (A) The codon wheel depicts the 64 codons that encodes...https://doi.org/10.1038/nsmb1031 Gustafsson C, Govindarajan S, Minshull J (2004) Codon bias and heterologous...3645–3650 https://doi.org/10.1073/pnas.0909910107 Varani G, McClain WH (2000) The G·U wobble base pair. ...

...PMID: 18478103. PubMed Central PMCID: PMC2364656. 2. Dittmar, Kimberly A., et al. "Selective charging ...PMCID: PMC552224. 4. Gustafsson, Claes, Sridhar Govindarajan, and Jeremy Minshull. "Codon bias and heterologous...: 346-353. PubMed PMID: 15245907. 5. Maertens, Barbara, et al. "Gene optimization mechanisms: A multi‐...

...Henrietta Lacks. New York: Crown Publishers, 2010. 2. Gold, Michael. "A conspiracy of cells." State University...line identity and purity making it easier for comparable studies to be performed on a cell line even decades...preferably some of these vials will be stored in a separate backup liquid nitrogen store to prevent loss from...