...relatively pH-insensitive RFP variant, DsRed.T3 and a pH-sensitive GFP variant, super ecliptic pHluorin (...trickier to identify because their contents can be at various stages of degradation. Additionally, electron microscopy...sequence. Under normal growth conditions, both variants are excluded from the vacuole and emit red and...the cytosol-targeted and mitochondria-targeted variants are available from Addgene. References 1. Rosado...

...Nabet (behnam_nabet@dfci.harvard.edu), Nathanael Gray (nathanael_gray@dfci.harvard.edu), or Jay Bradner ...an FKBP12F36V binding ligand (ortho-AP1867) via various chemical linkers. Our current lead dTAG molecule...Bradner (james.bradner@novartis.com). Considerations when getting the dTAG system up and running in your lab...While expression of these cereblon neo-substrates varies in different contexts (for example, IZKF1 and IZKF3...

... are created through enzymatic cleavage of the variable region (the antibody region that binds to the ...immunogenic version is needed.  Single-chain fragment variables (scFvs) are ~27kD engineered fusion proteins similar...instead of cleavage. They are comprised of the variable heavy and light chain regions, connected by a ...high-affinity RNA molecules can bind to a wide variety of targets, including proteins, peptides, amino... used either with or as fluorophores in a wide variety of colors.  To antibody or not to antibody?  That...

...mosaicism would occur. Although Ma et al. looked at various predicted off-target effects, they limited their... target. In addition, as on-target editing rates vary widely from gRNA to gRNA, it’s unclear where this...ethics committees comprised of individuals with varied backgrounds. Talking about this study with non-... Like me, you’ve probably seen this study (and various articles and commentaries) pop up on Twitter and...CRISPR Topic Page Read about High Fidelity CRISPR Variants Target Multiple Genes at Once with CRISPR Resources...

...co-factors, thus allowing for broad applications in a variety of experiments. LoxP (locus of X(cross)-over in...have been constructed that express Cre under a variety of ubiquitous and regulated promoters, and many... "NNNTANNN", where "N" indicates which bases may vary from the canonical sequence. Among these are loxN...lox2272 (GgATACtT), and lox511 (GtATACAT). These variant lox sites undergo recombination with other sites...not cross-compatible. Using different lox site variants allows Cre to catalyze more than one specific ...

...modified ABE7.10 accordingly which resulted in variants named “ABE8.x”: “8” stands for the eighth round...x-d.  This resulted altogether in 40 new ABE8 variants. When comparing on-target DNA editing efficiency...replaced S. pyogenes Cas9 in the ABE complex with PAM-variant Cas9 proteins: the S. pyogenes NG-Cas9 (PAM: NG...PAM: NNGRRT) to create Sa-ABE8.x-m/d. With these variants, ABE8 could target many additional locations in... base editor: ABE8e Read about xCas9, a CRISPR variant with PAM flexibility Resources on Addgene.org ...

...COVID-19 research in other labs. Spike protein, variants, and pseudotyping The SARS-CoV-2 virus depends...cellular receptor. To more easily study the spike variants, scientists have developed ways to pseudotype ... has had many opportunities to evolve. Several variants are now circulating and plasmids containing these...pseudovirus and those encoding different spike protein variants that can be incorporated into pseudoviruses. The...at neutralizing the different SARS-CoV-2 spike variants. You can learn more about their research in this...

...director of new initiatives at pgEd, I take on a variety of projects in the space of science education, ...genetics pgEd, based at the Department of Genetics of Harvard Medical School, has a mission of raising awareness...ethnic or personal identity. We do this through a variety of activities, including creating educational materials... disciplines, and everyone on staff takes on a variety of projects. With our focus on engaging the community... materials or speak about our experiences in a variety of other settings, including “conventional” academic...

...discussion on effective advocacy strategies on various levels, with panelists including: Kate Stoll, ...Project Lisa Coleman, PhD - Chief Diversity Officer Harvard University  The symposium will culminate with ...appropriate audience and communicating that message in various ways. Workshops will be run by experts in their...Cameron, PhD - Director of Media Communications, Harvard University Ray Howell -Founder of Howell Communications...David T. Riglar. David T Riglar is a postdoc at Harvard Medical School. In the lab, he uses synthetic biology...

... its variants allow for inhibition of neurons using blue-green light. Mac Variants. Mac variants have ...Inhibitory (Hyperpolarizing) ChR Variants. Alternatively, ChR variants that inhibit neurons have been created... these variants result in the hyperpolarization of neurons. Examples of anion channel variants from other...thus inhibiting function of the neuron. NpHR Variants. Variants have been engineered with enhancements such... triggered by green-yellow light. Arch Variants. Arch variants have been developed with the following ...plasmids . Glossary of Microbial Opsin Variants Opsin Type Variant Description Peak Response Spectra (nm...Phobos Blue-shifted iC++ variant 467 Aurora Red-shifted anion-selective ReaChR variant 517 Halorhodopsins Jaws...

...downstream applications. As individual libraries can vary significantly, it’s important that we have an amplification...unique plasmids does your library contain?), the variance of size of plasmids in your library (Are the inserts... cells you need total and per transformation may vary depending on the particular type of cells you are...plasmids to overtake larger plasmids, if the library varies significantly in plasmid size). On plates, each... Library Type Name Number of Plasmids Variable Region Size DNA per Amplification Amplification...

...in a variety of applications including gene expression profiling, studying copy number variation, and ...nucleotide concentration, unlike qPCR where results can vary across replicates. ddPCR can be used to quantify...concentration Gene expression profiling Copy number variations Quantify RNA levels Molecular Diagnostics NGS...of starting material AAV Titrations Allelic Variations Quantifying low abundance targets Monitoring ...

... Throughput Low Low Low Efficiency Variable Variable Variable Difficulty Hard Hard Hard These ...system of choice. The table below summarizes the various components included with each expression system...tunable expression of Cas9 and/or gRNA in a wide variety of mammalian cell lines. Useful for difficult to...

...the genetic elements may differ (e.g., a set of various mammalian promoters), the same cloning procedure..., titrate inducible transgene expression, or a variety of other clever applications. In one example, ...downstream applications. The tradeoffs between various cloning methods are listed in Table 2. For example...representation. The specific restriction enzymes used in various chaining-based cloning methods are listed in Table...MXS-chaining method have used the technique to engineer various constructs with applications in flow cytometry ...

...immunoglobulins, the strength of the interaction varies between antibody host species and isotypes. Consequently...factors to consider. Agarose beads are sponge-like and vary in shape and structure. Their porous surface provides...immobilized. The ideal antibody concentration to use will vary between capture antibodies and can be determined...target protein can be eluted. The elution method will vary depending on the specific target protein:capture... of interest on the C- or N-terminus. In this variation, called a pull-down assay, an antibody against...

...at all in E. coli (you can find a variety of databases showing various organisms’ codon preferences online...have different codon usage biases? The reasons for varied codon preferences among organisms aren’t completely...different levels under different conditions. With varied codon usage, an organism can change which proteins...the commercial Rosetta E. coli strains express a variety of tRNAs that are normally found at low abundance...

...either transcript variant led to robust and specific upregulation of the targeted variants without inducing...activation? To summarize, CRISPRa uses a nuclease-dead variant of Cas9 (dCas9) and engineered combinations of ...inducing expression of the non-targeted variant. Moreover, isoform-specific expression of Bdnf resulted in... between the Bdnf I and Bdnf IV overexpression variants were other immediate-early genes associated with...

...Mycoplasma contamination has been shown to arise from a variety of sources such as serum, other cell lines, or ...freeze-thaw cycles on lentiviral stability depends on a variety of factors such as the transgene expressed, the..., this study found that transduction efficiency varied not only between serum from different manufacturers...produce lentivirus may want to consider testing a variety of cation and serum sources to ensure the best ...applications, stick with them! Changes in such variables as cell density, volume, incubation time, and ...

...transpeptidase sortase A variant (mgSrtA), which is capable of promiscuously labeling various cell surface proteins...suitable for organoid or in vivo studies (Che and Khavari, 2017). Find APEX plasmids at Addgene! Since the...a small peptide (LPETG). In EXCELL, the mgSrtA variant can be displayed on the surface of a cell of interest...Central PMCID: PMC6126969. Che, Yonglu, and Paul A. Khavari. "Research Techniques Made Simple: Emerging Methods...

...by Jacob Lazarus, a postdoctoral researcher at Harvard. There’s an astounding number of ways to create...there is also a large amount of user-to-user variability, and in some cases, counter-selection escape ...thanks to our guest blogger Jacob Lazarus from Harvard. Jake received his MD and a PhD in Cell and Molecular...post-doctoral research in the lab of Matthew Waldor at Harvard. Now that he has an efficient allelic exchange ... PubMed Central PMCID: PMC3748948.  Khetrapal, Varnica, et al. "A set of powerful negative selection systems...