...properly exiting the ER and fluorescing (see Figure 2) [5].  When using FPs containing two cysteine residues...compartments including the secretory pathway (Figure 2), the inner membrane space of mitochondria and chloroplasts...PMID: 18228466. Pubmed Central PMCID: PMC2875081. 2. Snapp, Erik Lee. "Fluorescent proteins: a cell biologist's...dark fusions is likely misfolded, blocked from proceeding through the secretory pathway, and retained in...

...that the signal to noise ratio can be increased from 2-fold (by CRISPRainbow) to 3-fold with CRISPR-Sirius...CNV) analysis of the U2OS cells.   Figure 2: CRISPR-Sirius tricolor labeling. (A) Diagram of sgRNA-Sirius...and how these arrangements may affect nuclear processes. The latest of these visualization tools is CRISPR-Sirius...labeling of chromosomal loci in human cells.” Proceedings of the National Academy of Sciences of the United...

...while increasing the standard mutation rate by only ~2-fold over nCas9 levels. Two sets of modifications ...10-3 mutations per nucleotide per generation, and (2) the addition of the thioredoxin-binding domain (TBD... the nick site, which was expected since the processivity window of PolI is 15-20 bp. EvolvR’s on-target...bacteriophage T7 DNA polymerase increased the processivity of PolI3M, and thereby increasing the editing...

...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...easier with more perspectives contributing to the process Senior people (of which there are never enough,...during the meeting.  Scientists do better with process, so don’t just get together and chat. Have an agenda...started. Have regular meetings. Experiment with the process. Have each person write a Development Plan. Set...

... to share information about the emerging SARS-CoV-2 virus quickly and openly. By sharing results faster...Open Scientific Communication Ecosystem Fig. 2: The scientific ecosystem, enriched by open access...These each have different goals, formats, and processes, and they provide different amounts of access ...journal-based peer review is usually a confidential process among a few individuals.  Early-career researchers...errors, with a lack of transparency in most review processes. Preprints and their feedback, in contrast, are...

...recognize Fab fragments from the same species and 2) these fragments will be more likely to bind (and ...epitopes recognized by the secondary.   Figure 2: There are a few common solutions to avoid background...Created with biorender.com. Proper tissue processing and sample preparation can also help reduce the...

... (1) the struggle between two opposing forces or (2) the result of competing desires or (3) the presence... an introduction, and a resolution.   Fig. 2: The structure of a story - not just for fiction! ...find it within the science itself - within that process of failing, over and over again until you don't...

...panning and scFv characterization.    Figure 2: Phage display starts with a pooled library of scFvs...the short epitope GCN4, also called a scaffold; and 2) a GCN4 binding scFv that’s fused to GFP. When the...Fv (Variable Domain) (scFv) Fragments. Antibodies 2:193–208 . https://doi.org/10.3390/antib2020193  Khantasup...is displayed on the coats of bacteriophages, a process which links genotype and phenotype. Incubating ...single-chain Fv analogue produced in Escherichia coli. Proceedings of the National Academy of Sciences 85:5879–5883...

...11 and Cas13 are 1). non-specific RNA cleavage and 2). cell toxicity. Cas7-11 does not display collateral...Cpf1 is a single RNA-guided endonuclease of a class 2 CRISPR-Cas system. Cell, 163(3): 759-771 (2015). 10.1016...complementary site in the genome where it will proceed to generate a blunt DNA double strand break. Cas9...stranded DNA degradation. Cas3 DNA digestion can proceed up to multiple kilobases, but only in one direction...) and does not require a tracrRNA (it can self-process pre-crRNA into crRNA). The size and simplicity ...11 cuts ssRNA and is capable of pre-crRNA self-processing. Functionally, it is very similar to Cas13 and...

...the nuclear export and translation processes as well.   Figure 2: The conserved eukaryotic polyadenylation...due to the presence of additional helper sequences (2-3). As alluded to above, termination and polyadenylation...transcriptional unit (such as a gene) and initiate the process of releasing the newly synthesized RNA from the...terminator also plays an important role in RNA processing and contributes to variability in RNA half-life...Although mostly thought of as a eukaryotic-specific process, prokaryotes also add poly(A) tails to certain ...studied as prokaryotic termination, the basic processes for eukaryotic termination are understood and ...to terminate transcription. Polymerase I uses a process similar to the prokaryotic Rho-dependent mechanism...

...parts: (1) β-strands 1-9 (β1-9) and the one ɑ-helix, (2) β-strand 10 (β10), and (3) β-strand 11 (β11). Each...activating caspase-3, and detected fluorescence within 2 to 5 hours, a time consistent with previous methods...plays a pivotal role in an array of biological processes including development, the immune system, and ...cell turnover. Apoptosis is a highly controlled process that is triggered by internal and external signals...

...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...the importance of understanding these complex processes can’t be overstated, we can also learn a lot from...

...represent the preexisting resistant subpopulation 2. Resistant cells among the separate replicates would...medicine 21.5 (2015): 440-448. Pubmed PMID: 25849130. 2. Hata, Aaron N., et al. "Tumor cells can follow distinct... she could “get exposure to the drug discovery process.” The ClonTracer library originally came about ...that these are selected for during the treatment process. The authors went on to show that combination treatment...

... pCRISPomyces-2 contains Cas9 and a gRNA cassette. The simpler system of pCRISPomyces-2 displays a higher...and a subsequent curing protocol that requires only 2-3 hours incubation. E. coli and T. citrea scarless...four major components including (1) a CRISPR RNA, (2) four proteins forming the QCascade DNA-targeting ...inefficiency? Use CRISPR to make it a selectable process! As NHEJ is ineffective in bacteria, CRISPR-induced...collection of organisms, their genes, and metabolic processes, occupying the gut. Many human diseases have connections...properties of CRISPR-Cas systems is base editing. This process is generally accomplished by fusing an enzyme capable...

...host range plasmids. FEMS Microbiology Letters, 348(2), 87–96. https://doi.org/10.1111/1574-6968.12241  ...mobilization of broad host-range IncQ plasmids. Plasmid, 62(2), 57–70. https://doi.org/10.1016/j.plasmid.2009.05.001...negative hosts. Biotechnology and Bioengineering, 106(2), 326–332. https://doi.org/10.1002/bit.22695  Schmidhauser..., as they allow for simplified transformation processes, and can be used by scientists across disciplines...

...the reporter mRNA is labeled by PCP-mCherry (Figure 2). The 3’ UTR also contains a CAAX sequence to bind...PMC2746483 Find ribosome profiling plasmids at Addgene. 2. Halstead, James M., et al. “Translation. An RNA biosensor...useful, this technique requires extensive sample processing to avoid disrupting ribosome-mRNA complexes. ...

...nonsense-mediated decay. Although Nelson et al. observed only 2% genome editing in one experiment, they found the ... cells throughout the body, especially stem cells 2. Remove any immunogenicity of the AAV vector Once ...Science. 2015 Dec 31. doi:10.1126/science.aae0169 2. Long C, Amoasii L, Mireault AA, McAnally JR, Li H... mature tissues - no problem, as exon skipping proceeds through the non-homologous end joining (NHEJ) ...is upregulating HDR in mature tissues, as this process is necessary to precisely edit point mutations....

... you can read more about them in review articles (2–4). You can also take a look at my recent paper to...Development 118, 401–15 (1993). PubMed PMID: 8223268. 2. J. B. Duffy, GAL4 System in Drosophila : A Fly Geneticist...post, I’ll explain the relatively straightforward process of generating your own fly lines. In the meantime...

...is always best to have options.      Fig. 2: Addgene’s western blot for mEGFP-HRas (from Plasmid...the presence of oxidizing agents. Adapted from Fig. 2 Campbell et al, Nature Methods   Find YFP plasmids...exposure can negatively affect FPs during these processes. The Liu and Petsko labs recently engineered hyperfolder...

...Interview with Stem Cell Researcher Darrell Kotton 2. Stem cell scientists care about patients As any grad... Yamanaka explained that it took 50 people nearly 2 years to create the first stem cell line that could...be used in clinical trials. The quality control process alone takes nearly a year. Addgene depositor Juan...