We narrowed to 492 results for: ara-2
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TypeBlog Post...amilCP, tsPurple, and gfasPurple) (Figure 2). Figure 2: Chromoprotein expression from bacteria on.... https://doi.org/10.1007/bf00428249 Chan MCY, Karasawa S, Mizuno H, Bosanac I, Ho D, Privé GG, Miyawaki...Miyawaki A, Ikura M (2006) Structural Characterization of a Blue Chromoprotein and Its Yellow Mutant from ...Hoegh-Guldberg O (1995) Isolation and Partial Characterization of the Pink and Blue Pigments of Pocilloporid...
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Viral Vectors 101: Voltage Indicators
TypeBlog Post...Indicators GEVIs can be divided up structurally into 2 main groups: voltage sensitive domains and Rhodopsin...depending on your experimental system. Figure 2: General overview of chemigenetic tools. A genetically...mechanism of their voltage sensing component, which separates them into two distinct categories detailed below...phosphatases from sea squirts. This design structurally separates the two components of the indicator, which can...FP-rhodopsin GEVIs have been engineered to have kinetics comparable to its one component counterpart and is significantly... -
Deep Dive: Fixing and Permeabilizing for Immunofluorescence
TypeBlog Post... of permeabilization and membrane integrity (Fig. 2). For example, saponin interacts with cholesterol ... at too high a concentration. Figure 2: Examples of different permeabilization strategies...cause some confusion. Paraformaldehyde: Many IF protocols call for paraformaldehyde (PFA) as the fixative...recover good antibody binding by tweaking those parameters, but in some cases your antibody may just not... -
Fluorescent Proteins 101: Green Fluorescent Protein (GFP)
TypeBlog Post...from GFP and the mutation definitions found in Table 2 may not apply. Table 1: The specific mutations comprising..., V163A EBFP F64L, S65T, Y66H, Y145F Table 2: Functional role of specific mutations in GFP derivatives... color also exist with only slight variations separating them. Please note that many fluorescent proteins...(FACS): This is a type of flow cytometry that separates mixtures of cells into distinct populations based... fluorescent signal. Thus, FACS can be used to separate cells expressing GFP from cells that are not. ... -
R Bodies: Membrane-Rupturing Microscopic Tools
TypeBlog Post...be decorated with proteins and remain functional, 2) break membranes outside of their natural context ...PubMed PMID: 3759909. PubMed Central PMCID: PMC213417. 2. Schrallhammer, M. et al. Tracing the role of R-bodies...toxicity of R-body producing recombinant E. coli on paramecia. European Journal of Protistology 48, 290–296 ... -
Pooled CRISPR Libraries Offer Genome-Wide Control for Large-Scale Functional Screens
TypeBlog Post...VP64 fusion; 2. A sgRNA incorporating two MS2 RNA aptamers at the tetraloop and stem-loop 2; 3. The MS2...Adamson B, Villalta JE, Chen Y, Whitehead EH, Guimaraes C, Panning B, Ploegh HL, Bassik MC, Qi LS, Kampmann... -
Advice for Starting a Biotech Company from a Grad Student turned Entrepreneur
TypeBlog Post...go anywhere. The team should initially consist of 2-3 individuals that are aligned in vision and outcomes... the technology. This transition takes around 1 – 2 years. The product is the measure of success. Technology...constantly practicing. “The expenditure of effort in preparation and practice is directly related to success presenting... -
Form Your Own Peer Mentoring Group: A How-To Guide for Scientists
TypeBlog Post...The groups in these programs typically consist of 1-2 mentors and 3-5 mentees who meet together for an 8... together. If you all work at the same place, a 1-2 hour lunch on a regular schedule might work. If not...pre-reads, videos or exercises to be completed in preparation for the meeting or to be read or watched during... -
Getting to Know Expansion Microscopy
TypeBlog Post...your sample (i.e. cultured cells or tissues). Step 2: attach anchors to your sample that will link biomolecules...quite so much. Created with biorender.com Steps 2-4 are not found in standard IHC/ICC protocols and ...to ~4-10x linearly! This expansion physically separates molecules while maintaining their relative positions...pre-expanded samples can sometimes take on a fragmented appearance post-expansion, but that doesn’t mean we should...understanding of neuronal synapses, centrosomes, parasites, and more (Bertiaux, et al. 2021; Gambarotto, ...new insights into the cytoskeleton of malaria parasites including the conservation of a conoid. PLoS Biology...10.1038/s41592-018-0238-1 Klimas A, Gallagher BR, Wijesekara P, et al (2023) Magnify is a universal molecular... -
Tips for Screening with Yeast Two Hybrid Systems
TypeBlog Post...domains retain their individual activities when separated (2), and that DBD and TAs from different systems...308. PubMed PMID: 6312261. 2. Keegan, L., Gill, G., and Ptashne, M. Separation of DNA binding from the transcription-activating...PubMed PMID: 2547163. 5. Ozawa, T., Kaihara, A., Sato, M., Tachihara, K., Umezawa, Y.l. Split luciferase...possibility for problems with one or more of these parameters can lead to a high number of false negatives ... -
Antibodies 101: Polyclonal Antibodies
TypeBlog Post...animal is given additional booster immunizations every 2-3 weeks to increase the antibody titer. The titer ...level of antibodies is reached - typically within 2-4 months. After the animal has the desired level of...animal. The red blood cells and the serum are separated, and the serum - which contains the antibodies... -
INTEGRATE: Bacterial Genome Engineering Using CRISPR-Transposons
TypeBlog Post...et al., 2020). Genomic targets require an upstream 2-bp PAM sequence for recognition by QCascade; although...cells of the plasmid after integration. Figure 2: (A) Streamlined VchINT expression constructs. (B)...target DNA sequences using RNA-DNA base pairing. Separately, TnsABC forms a transposase complex with the ...bp target-site duplication (TSD), a hallmark characteristic of the Tn7-family of transposons. Insertion...Sternberg group’s initial report of VchINT, three separate plasmids and multiple promoters were used to express...Additionally, by expressing multiple crRNAs in parallel (Fig. 3), multiplexed insertions into multiple... -
Empty Backbones - Choosing Your Perfect Plasmid Backbone
TypeCollection...backbone with MS2 loops at tetraloop and stemloop 2 and EF1a-zeo resistance marker pLenti CMV/TO GFP-Zeo...Tet) Inducible Expression page Bacteria Lac, T7, araBAD, trp mTagBFP2-pBAD - Protein expression vector ...astrocyte-selective expression in AAV Dual promoter Separate expression pRGEB31 - Deliver sgRNA and Cas9 into... -
CRISPR 101: Cytosine and Adenine Base Editors
TypeBlog Post... a template, copying the base edit. Figure 2: Cytidine deamination takes place on the free strand...SpCas9 base editors with editing windows as small as 1-2 nucleotides. To reduce off-target effects associated...formation. To further decrease indel formation 1.5-2 fold, the team fused bacteriophage protein Gam to ...which we've discussed in two follow-up posts: (1) (2). Base editors have a narrow editing window of only...K., Arazoe, T., Yachie, N., Banno, S., Kakimoto, M., Tabata, M., Mochizuki, M., Miyabe, A., Araki, M.,...., Slaymaker, I. M., Yen, J., Young, L. E., & Ciaramella, G. (2020). Directed evolution of adenine base...Schwaegerle, N. D., Peraro, L., Young, L., Lee, S.-J., Ciaramella, G., & Gaudelli, N. M. (2023). Improved cytosine... -
CRISPR 101: Epigenetics and Editing the Epigenome
TypeBlog Post...form or as ready-to-use lentivirus. Figure 2: dCas9-Tet1 demethylates cytosines at promoters and...pX330a dCas9-LSD1 for targeted removal of H3K4me1/2 and H3K9me2 histone modifications. Like the TALE-LSD1...approach for targeted DNA demethylation. Cell Discovery 2: . https://doi.org/10.1038/celldisc.2016.9 Find plasmids...could identify the target genes of previously uncharacterized enhancers. The popular TALEN-based LITE system... -
A New Generation of Adenine Base Editors Improves Editing in Primary Human Cells
TypeBlog Post...NGG-3′ (“N” can be any nucleotide base, followed by 2 guanines). This limits the use of ABEs as DNA sites... in the promoter region of gamma hemoglobin (HGB1/2) leads to continuous expression of fetal hemoglobin...found that the smaller version, ABE8.x-m, showed comparable editing behavior to ABE8.x-d. Overall, ABE8s ...MS, Rinaldi C, Slaymaker IM, Yen J, Young LE, Ciaramella G (2020) Directed evolution of adenine base editors... -
Magnetic Control of Proteins: More than a Dream
TypeBlog Post...of ~75% in response to the magnetic field (Figure 2); they named this variant MagLOV. Andrew was thrilled...didn’t need any added cofactors. Figure 2: Fluorescence intensity and ΔF/F of MagLOV, a variant... that don’t even exist yet? Andrew York, Maria Ingaramo, and their team at Calico Life Sciences recently...across. Video courtesy of Andrew York and Maria Ingaramo. MagLOV shows a wealth of potential, and while...Lazzari-Dean, J. R., Lefebvre, A. E. Y. T., York, A. G., Ingaramo, M. (2024). Magnetic control of the brightness... -
Fluorescent Proteins 101: Fluorescent Protein Timers
TypeBlog Post...linear region to solve for total expression time. 2. Timer functionality with a single gene Although it...290.5496 (2000): 1585-1588. PubMed PMID: 11090358. 2. Tsuboi, Takashi, et al. "Age-dependent preferential...possible to make a timer system composed of two separate FPs (one that matures quickly and the other slowly...s directed mutagenesis also produced the three separate timers shown in the table: Fast-FT, Medium-FT,...varied blue-red maturation times (Table 1). These separate proteins should be useful for monitoring cellular... -
Deciphering the Mysteries of Behavior with Viral Vectors
TypeBlog Post...of these neurons via the channel Channelrhodopsin 2 (ChR2) reduced glutamate transmission from the mPFC...deliver Cre-dependent expression of Channelrhodopsin 2 (ChR2), a modified receptor protein that activates...movement in an open field box or a circular corridor apparatus using a camera or tracking software. Locomotion...the open (risky). Taking advantage of these characteristic traits, rodents can be used to study anxiety...measuring the time spent in a particular part of the apparatus, it turned out that the IL-1 beta overexpressing... -
GPCRs: How Do They Work and How Do We Study Them?
TypeBlog Post...plasmid kit consists of 20 plasmids encoding 14 alpha, 2 beta, and 4 gamma subunits of the heterotrimeric G...activation via bioluminescence resonance energy transfer 2 (BRET2). Check out our blog post on GPCR signaling...created an open source resource called PRESTO-TANGO (parallel receptorome expression and screening via transcriptional...