...article in the Addgene Blog Mentoring series will cover 2 of the 7 mentoring questions I set out to answer. ...101-Building Relationships for Success”. Choose 1-2 tactics to try. One of my favorites is making the ...ardor and attended to with diligence.” - Abigail Adams, 1780 There are potential mentors all around you...

... 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 ...systems to generate ABE8e(TadA-8e V106W), which edits ~590-fold faster than the TadA from ABE7.10 without ...Engineered TadA domain for cytosine base editing Lam et al., 2023 CBE-T, CABE-T   Engineered TadA domain...evolution to create one from the RNA adenine deaminase TadA.   After seven rounds of molecular evolution, they...approach, engineering and directed evolution of the TadA domain has yielded base editors that are equally...

...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...MagLOV’s parent protein AsLOV2 has already been adapted into a variety of optogenetic tools, they’re optimistic...optimistic that MagLOV can be similarly adaptable into future magnetogenetic tools.  Embracing science...

...double strand break), increasing editing efficiencies 2-3-fold. However, the process of double nicking increases... editing as efficient as possible. Figure 2: Decision tree for selecting a PE6 variant. PE = prime...using paired prime editing. Nature Biotechnology, 40(2), 218–226. https://doi.org/10.1038/s41587-021-01025...Traditional sgRNAs are protected from cellular degradation by Cas9 binding, but pegRNAs have an extended... RNA stability This recent contribution by the Adamson lab also addresses the problem of RNA stability...A., Sahin, M., Osborn, M. J., Weissman, J. S., Adamson, B., & Liu, D. R. (2021). Enhanced prime editing...., Bauer, D. E., Marson, A., Parsons, L. R., & Adamson, B. (2024). Improving prime editing with an endogenous...

...antibiotic selection. Essentially, this gene is a 2-in-1 providing both bacterial and eukaryotic selection...Zeocin-induced DNA cleavage in human cells. Mutagenesis, 20(2), 111–114. https://doi.org/10.1093/mutage/gei016I ...resistance to both carb and amp, due to their common degradation mechanism. How to choose between the two? Carb...high pH environments and is less susceptible to degradation in agar plates. This prevents satellite colonies...

...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)...spreading within and between genomes, and have been adapted for a wide variety of applications - as mutagenesis...at two different incubation temperatures. Data adapted from Vo et al., 2020. In E. coli, the new streamlined...

...connectivity to other data sources used in research; (2) connectivity to other tools used in research; and...by making it easy to get data out of the ELN; and (2) facilitate inter-operability with other research ...support entry of required metadata, with the option of adding additional metadata in the repository after...

...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...last in the cascade and are responsible for the degradation of over 600 cellular components ultimately leading...

...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...were known to be resistant to detergents and degradation under a variety of conditions. They are composed...

...PubMed Central PMCID: PMC3785459. 2. Krashes MJ, Koda S, Ye C, Rogan SC, Adams AC, Cusher DS, Maratos-Flier... tropism and axonal transport of AAV serotypes 1, 2, 5, 6, 8 and 9 in the mouse brain. PLoS One. 2013 ...

...VP64 fusion; 2. A sgRNA incorporating two MS2 RNA aptamers at the tetraloop and stem-loop 2; 3. The MS2...Repression and Activation. Gilbert LA, Horlbeck MA, Adamson B, Villalta JE, Chen Y, Whitehead EH, Guimaraes...

... 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...asapbio.org/preprints-and-covid-19 Ettinger CL, Sadanandappa MK, Görgülü K, Coghlan KL, Hallenbeck KK, Puebla...

...wheat germ, HeLa, and Leishmania (Gagoski, et al 2015[2]). The proteins encoded in this set range in size ...to generate Illumina sequencing libraries using a 2-step PCR-based protocol and are compatible with single-end...populations. This kit consists of a 48 plasmid adapter library, with each plasmid carrying a unique barcoded...selective pressures: Miller, Corwin, et al. "Adaptation of Enterococcus faecalis to daptomycin reveals...lead to surprisingly large fitness effects during adaptive evolution of antibiotic resistance." Proceedings...

...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...low-frequency indels will not cause frameshifts. Figure adapted from Osterout et al. (2015b) under a CC-BY 4.0 ...

...without leaving behind any accessory DNA sequences [2], [3]. Simply put, the piggyBac™ transposon containing...with a Footprint-Free™ genome edit.   Figure 2: Editing DNA using the piggyBac™ transposon selection...PMID: 26211958. PubMed Central PMCID: PMC4663986. 2.  A. M. Singh, V. V Adjan Steffey, T. Yeshi, and D.... Jonkers, M. van Lohuizen, A. Berns, and D. J. Adams, “A high-throughput splinkerette-PCR method for ...

...elution choices: TE, Tris buffer, or water Figure 2: Tris-EDTA, Tris, or water? The buffer you choose ...product, or chromosome. One of the main causes of degradation during plasmid DNA extraction and purification...protein endonuclease I encoded by the gene endA. Nowadays, the most common E. coli strains used for plasmid... extra steps like heat inactivation, protein degradation, and/or the use of chelating agents that can ...store your sample without having substantial degradation and which experiments you can use your plasmid...

...might otherwise be hard to identify. Figure 2: Users can cross-interrogate screens from the literature...relevant metadata and the screen results. Screenshots updated 2024. Importantly, metadata for each...gene hits for each screen are also captured as metadata.   Screen results are reported as the original... diversify, the BioGRID ORCS resource will be adaptable to a wide variety of screen formats and contexts...

...educational background and position at Novartis. Chapter 2: What's a lab group in industry? The collaborative...and leveraging your unique skills set while also adapting to needs as they arise. Tips on applying for industry...

...Hrit et al. 2018, Kaas et al. 2013). Figure 2: CRISPR TET-based epigenetic editing. In reality, ...approach for targeted DNA demethylation. Cell Discov 2, 16009. https://doi.org/10.1038/celldisc.2016.9.  ...methylation, leaving the target CpG unmethylated. Adapted from Sapozhnikov and Szyf 2021.   Let me leave...

...is funded have been well documented over the past 2 years (Alberts et al, PNAS 2014; Taylor, eLife, 2016...PMID: 24733905. Pubmed Central PMCID: PMC4000813. 2. Taylor, Justin Q., et al. “Avoiding a lost generation...Office Ben Corb - Director of Public Affairs at ASBMB Adam Fagen, PhD - Director, Early Career Scientist Segment...