We narrowed to 769 results for: IRE
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TypeBlog PostPublishedNov. 12, 2020, 2:15 p.m.... the ideal codon for every amino acid across an entire polypeptide. Many researchers have created codon-optimization...necessary for proper protein folding (Stadler and Fire, 2011). Therefore, codon optimization may disrupt...1987) The codon adaptation index-a measure of directional synonymous codon usage bias, and its potential...doi.org/10.1016/s0165-0270(02)00362-x Stadler M, Fire A (2011) Wobble base-pairing slows in vivo translation...
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Harnessing TnpB for Genome Editing: A Compact and Versatile Tool for Your Research
TypeBlog PostPublishedJuly 29, 2025, 1:15 p.m....cleave both strands. Similar to how Cas proteins require a PAM (protospacer adjacent motif) near the target...a different TAM (5′-TTAT-3′). The unique TAM requirements expand the number of genomic sites that can ... and prime editing (Molla et al., 2021), which require fusing Cas proteins with additional enzymes or ...transformation into plant cells. Conclusion & Future Directions TnpB represents a new generation of compact, ... -
QC Sequencing Technologies at Addgene
TypeBlog PostPublishedJuly 15, 2025, 1:15 p.m....strand is synthesized (Figure 1). Sanger sequencing requires four separate reactions, where a fraction of one...in ultra-long read mode), enough to sequence an entire plasmid in one read. Runs can be completed in anywhere...sequencing, as well as the ability to sequence an entire viral genome in one read. We also use Nanopore ...keeping up with the large amount of QC analysis required every week. They are constantly exploring ways... -
Plasmids 101: Shuttle Vectors
TypeBlog PostPublishedMay 20, 2025, 1:15 p.m....helps increase cellular uptake, but this approach requires compatible cell lines that express viral machinery...ribosome. One resistance gene on your vector might require two different selection molecules! It all depends...option is to use a different method of selection entirely. Auxotrophic selection relies on the host organism...organism being unable to synthesize a nutrient required for growth. Before delivery of the vector, the deficient... -
Transferable Skills Guide: Creativity
TypeBlog PostPublishedJan. 18, 2022, 2:15 p.m....skill developed during PhD training. Creativity is required to develop new projects and test novel approaches... all the funding in the world? What if you could hire more people? What would you do with unlimited time...over half of which, when implemented, impacted the entire company. It is important to note that the person...product team is an incredibly innovative group. Our entire company benefits from their policy! Perfectionism... -
Four Base Editing Reporters to Monitor and Enrich Editing in Real-time
TypeBlog PostPublishedJuly 7, 2020, 1:15 p.m....DNA strain which results in two DSBs that can be repaired by NHEJ restoring mCherry expression. Cells can...populations. To develop this method the lab was inspired by work that used an integrated GFP reporter that...fluorescent protein) upon CRISPR/Cas9 homology directed repair (HDR) (Glasser et al., 2016). For TREE ...different reporter proteins. Protein translation requires the start codon of AUG immediately downstream ... -
Plasmids 101: Plasmid Incompatibility
TypeBlog PostPublishedMarch 31, 2020, 1:15 p.m.... Antisense RNAs encoded on the plasmid either indirectly inhibit replication through inhibiting translation...translation of replication machinery proteins, or directly inhibit replication by binding to the origin of...sequences within the origin of replication and are required for replication in plasmids that contain them ...Kamruzzaman M, Shoma S, Thomas CM, Partridge SR, Iredell JR (2017) Plasmid interference for curing antibiotic... -
Plasmids 101: Expressing shRNAs for RNA Interference
TypeBlog PostPublishedJune 17, 2025, 1:15 p.m....composition of the RISC varies, but at the very minimum requires an Argonaute protein. This Argonaute protein is...shRNAs using plasmids The design of shRNA plasmids requires three important steps: Designing the shRNA sequence...This is much shorter than miRNA plasmids, which require additional flanking regions comprised of upstream...35(9), 872–878. https://doi.org/10.1038/nbt.3947 Fire, A., Xu, S., Montgomery, M. K., Kostas, S. A., Driver... -
Viral Vectors 101: Integrase-Deficient Lentiviral Vectors
TypeBlog PostPublishedMay 13, 2025, 1:15 p.m....Ciuffi, 2012). One family of viruses, retroviridae, require integration for their life cycle to continue. They...IN (integrase). These viral packaging genes are required for the production of lentiviral particles. The... keep in mind is that titering these IDLVs may require a slightly different protocol than regular lentiviral... (2008). Insertional mutagenesis combined with acquired somatic mutations causes leukemogenesis following... -
Hot Biosensors 2022: Year-End Roundup
TypeBlog PostPublishedDec. 6, 2022, 2:15 p.m....variety of behaviors, opening up new research directions that were inaccessible with earlier tools. They... in localization to the cell membrane and not a direct change in the fluorescent properties, pYtags are...variations and heat transfer at the subcellular level requires a probe with fast kinetics and high signal/noise...temperature indicator B-gTEMP designed to meet those requirements (Lu et al. 2022). By fusing two bright fluorescent... -
Antibodies 101: Single Chain Fragment Variables (scFvs)
TypeBlog PostPublishedJune 3, 2021, 1:15 p.m....be expressed in bacteria, while mAbs generally require mammalian expression systems. scFvs are also small...to avoid animal use by using semi-synthetic or entirely synthetic VL and VH sequences. Phage display also...for the scFv (Gaciarz and Ruddock., 2017), but requires an oxidizing environment to form. When expressed... age-related macular degeneration (AMD) since direction injection to the eye results in high drug concentrations... -
Four Ways to Package Transgenes That Exceed the Size Limit of Adeno-associated Virus
TypeBlog PostPublishedSept. 1, 2020, 1:15 p.m....AAV donor template for CRISPR-mediated homology directed repairs (HDR). Split AAV vector approaches Split...trans-splicing (62%) vector. 4. Sequential homology directed repairs Overview: The sequential homologous recombination... donor templates using two sequential homology directed repairs (HDR) with CRISPR/Cas9. Donor A contains...M, Colella P, Garcia-Hoyos M, Gibbs D, Kim SR, Maguire A, Rex TS, Di Vicino U, Cutillo L, Sparrow JR, ... -
Multiple Plasmids at a Low Price: Kits from Addgene
TypeBlog PostPublishedSept. 14, 2016, 2:30 p.m....bypasses the 5′ mRNA cap required for eukaryotic systems and instead directly engages the ribosome for... protocol and are compatible with single-end or paired-end read flow cells. FREQ-Seq is an open source... The kit contains all the variant PUF segments required to bind any 8 base RNA sequence. The segments ... -
Antibodies 101: Multiplex Immunofluorescence
TypeBlog PostPublishedMay 23, 2023, 1:15 p.m....secondary antibodies altogether and multiplex with direct IF, using primary antibodies that are conjugated...However, the primary-secondary workflow (a.k.a. indirect IF) is more flexible and efficient, since you ...primaries for whichever targets you like. Plus, indirect IF amplifies the signal since multiple secondary...design. Like selecting an antibody, some of the requirements are similar to standard IF. Here, we’ll walk... -
Plasmids 101: Dimers and Multimers
TypeBlog PostPublishedJan. 10, 2023, 2:15 p.m.... on as a dimer, depicted on the right, with the entire monomeric sequence duplicated in tandem, resulting... with large inserts (>7 kb)(Berg et al., 1989), direct repeats (Ribeiro et al., 2009), and at high copy...frequency plasmid recombination mediated by 28 bp direct repeats. Mol Biotechnol. 2008 Nov;40(3):252-60....Summers D. Timing, self-control and a sense of direction are the secrets of multicopy plasmid stability... -
Antibodies 101: The Basics of Western Blotting
TypeBlog PostPublishedMarch 8, 2021, 2:41 p.m....overnight. In contrast, a semi-dry transfer only requires enough buffer to saturate the sandwich and typically...without any transfer buffer. These systems, however, require you to purchase expensive system-specific sandwiches...on the antibodies and labels being used and may require some trial and error (Yang et al., 2012). Choosing... they provide qualitative measurements. If you require precise quantification of a protein, consider using... -
Finding Your Perfect Job After University
TypeBlog PostPublishedJan. 12, 2016, 3:30 p.m.... degree. I loved parasitology. However, when I enquired about roles in this field, most jobs needed at...understood why parasitology roles in the field require experience. Working in industry: Microbiology ...redundancies can be made within the company; new staff hired in anticipation of the contract suddenly have less...institutes. The lab at the University of Queensland required a scientist for a 6 month project to validate ... -
Mycoplasma Contamination: Where Does It Come From and How to Prevent It
TypeBlog PostPublishedOct. 8, 2020, 1:15 p.m.... to stain all DNA. Some amount of expertise is required in this process because result interpretation ...serum to be used for subculturing. Avoid using expired products or reagents that have been stored for ...cross-contamination. Make sure you keep all the required equipment and reagents inside the laminar airflow... Biotechnology. References Degeling MH, Maguire CA, Bovenberg MSS, Tannous BA (2012) Sensitive ... -
Typing CRISPR Systems
TypeBlog PostPublishedMarch 18, 2025, 1:15 p.m....systems has led to extreme sequence divergence that requires a multi-pronged analysis to sort effectively. ...efficiency when conducting edits using homology-directed repair. There are 10 subtypes of type V, named...select RNA transcripts, or be engineered to conduct direct RNA editing. There are four subtypes, A–D, with...surface-level function of each type could lead you to new directions in your research. We all have our preferences... -
SciComm with the Experts at Science in the News Part 1
TypeBlog PostPublishedMarch 16, 2017, 1:06 p.m....University. Effective science communication is required to promote public support for research and to ...graduate student at Harvard University, former Co-director of Science in the News, and member of the SITN...her PhD from Harvard University, is a former Co-director of Science in the News, and is a member of the...