...ampicillin resistance cassette and an origin of replication, which are not schematized. Sizes of fragments...validated and were chosen because of their broad applicability in different model systems. What’s the error...five months. When they tested those clones (in duplicate), 93% had the correct banding pattern in the restriction...correct. Because they created the assemblies in duplicate, they were able to confirm that at least one plasmid...

...search algorithm rather than pre-designed sets, so identical searches can return multiple answers. You might...of a lac operon dropped to 49% fidelity, with significant lot-to-lot variation. Additionally, this assembly...assemblies are possible, the two sets of data together indicate that 35-pieces is a reasonable upper limit for...Overhang Ligation Fidelity by T4 DNA Ligase and Application to DNA Assembly. ACS Synthetic Biology. https...

...its design changes for every new prime editing application. And optimizing its sequence is important, as... gRNA (Anzalone et al., 2019), disrupting its canonical structure and Cas9 binding (Nishimasu et al., ...other bases in this location bind to Cas9 with canonical sgRNA structure (right). Created with BioRender.com...structured motifs like mpknot, use the pegRNA Linker Identification Tool (pegLIT) to create linkers that minimize...

...and should be varied as it can have important implications for the fusion protein effectivity), and a fluorescent...to visualize DNA, an editor targets DNA (via catalytically inactive dCas9) to modify chromatin. A fluorescent...both spatiotemporal protein degradation and quantification of this depletion using imaging-based measurements...BioTag, which can be used in protein affinity purification studies.    Duong et al., Genetics 2021. https...

...BioRender.com In their recent ACS Synthetic Biology publication, Mary Dunlop’s lab generated a photosensitive...regions with Vivid (Vvd) photodimers. Upon the application of blue light, Vvd dimerizes bringing the C-terminal... Jesse Zalatan’s lab deposited plasmids for a chemically inducible CRISPRa system in yeast is mediated... channelrhodopsin (C1V1) targeting parvalbumin cortical interneurons is now available as a ready to use...

... first at Brigham and Women’s Hospital/Harvard Medical School and then Boston University (institutions...committees would not function without long distance communication and learning to work with people despite the...the distance improved my verbal and written communication skills. Volunteering on these committees was ...Chair then stepped down, I became interim Chair automatically as it was an understood expectation of any Vice-Chair...

...detect explosives and rapidly indicate their presence. They are genetically modified organisms, and GMOs...things more interesting, we are working in a new technical discipline that nobody understands (synthetic ...that survived the literature review, survived technical feasibility analysis, and was a marketable product...

...to the same principles listed here. It employs biological best practices. For example, it ranks guides ...with NGS data were: Crispresso CrispRVariants AmpliCan We chose to adapt Crispresso by Luca Pinello ...HDR. Optimal mutation in-depth One the most sophisticated features of CrispyCrunch is how it mutates guide...published on how to do it best. Scientists are typically advised to mutate the PAM to which the guide binds...

...your gene of interest is difficult to target specifically with a gRNA). Be sure to keep these techniques...sequences are exchanged between two similar or identical molecules of DNA. Gene targeting takes advantage...homology is required, but 6 to 14 kb of homology is typical for targeting constructs. In the example shown ...cells participating in development or other physiopathological events to which the chosen promoter responds...

...genome editing technology that allows precise modifications of DNA (replacements, small insertions, and ... Liu, 2023). This method uses a prime editor (typically a Cas9 H840A nickase fused to a reverse transcriptase... design of the pegRNA has been shown to play a critical role in the editing efficiency of prime editing...configurations can be used to introduce specific target modifications. Since this means we could design hundreds to...