...Patterson GH, Ting LM, Wang Y, Condeelis JS, Verkhusha VV. A photoswitchable orange-to-far-red fluorescent...proteins Newly developed photoconvertible FPs like PsMOrange and mEos add another layer of complexity to imaging...fluorescent protein, PSmOrange. Nat Methods. 2011 Jul 31;8(9):771-7. doi: 10.1038/nmeth.1664. PubMed. Find ...

...BA, Schnitzbauer J, Zhang W, Li GW, Park J, Blackburn EH, Weissman JS, Qi LS, Huang B. Cell. 2013 Dec 19...inter- and intrachromosomal dynamics, and how they change in response to the cell cycle or other stimuli....cells. Ma H, Naseri A, Reyes-Gutierrez P, Wolfe SA, Zhang S, Pederson T. Proc Natl Acad Sci U S A. 2015 Mar...

...indicate the amino acid change as well as any functional consequences of that change. For example, "Mutated...number of unique plasmids in a pooled library can range from a few hundred to millions. Pooled libraries...through Addgene. For example, you could enter "Feng Zhang (Addgene plasmid #52961)".   The vector type should...by Tyler Ford in January 2016. It was updated by Angela Holmes and Christina Mork in February 2024.  Additional...

... by integrating a SL-insensitive fluorophore (LSSmOrange) as an internal control and a SL-sensitive fluorophore...enzymatic receptor DAD2. SLs induce a conformational change of the receptor which modulates the fluorescence...fluorescent biosensor has a high sensitivity (50 nM-500nM range) and a high specificity for SLs in vitro, and they...acssynbio.0c00192   The CRISPR corner By Jennifer Tsang New CRISPR plasmids are always being added to the...

...to the distinct overhang patterns generated by the nickases; D10A creates 5’ overhangs and H840A creates...creates 3’ overhangs in a PAM-out design.  Exploring nickases for homology directed repair The potential ... benefit of using nickases for HDR is targeting range: using an individual gRNA with WT Cas9, repair levels...

...Isaacman-Beck J, Paik KC, Wienecke CFR, Yang HH, Fisher YE, Wang IE, Ishida IG, Maimon G, Wilson RI, Clandinin...doi.org/10.1101/788679 Lin R, Wang R, Yuan J, Feng Q, Zhou Y, Zeng S, Ren M, Jiang S, Ni H, Zhou C, Gong H,...

...Using video conferencing tools like Zoom or Google Hangouts is a great way to include researchers from different...person Presenting the results from my lab exchange project at the International Cancer Immunotherapy...other in person. One possibility is to do a lab exchange, where researchers spent time in each other’s ...performed the same way at different sites. I did a lab exchange as part of the collaboration I led. This gave ...

...in the CRISPR-Cas9 field – Jennifer Doudna, Feng Zhang, and Keith Joung. All Addgene depositors! Addgene...Addgene was mentioned specifically in Dr. Zhang's introduction. His willingness to share reagents so freely... are taking the "genetic code" to a new level, changing it from a puzzle to decrypt to a tool to be harnessed... tools – both deposited and distributed – have changed so dramatically in the past 4 years. I kept busy...

...for our first post, I thought it best to define language you’ll see throughout the posts and introduce ...still considered scientific communication if the language is aimed at a peer audience.  Note that lay people....      Jargon is field-specific terminology or language. This can refer both to terms that are only used...number of characteristics. Common ones include age range, familiarity with the subject, shared interests,...

...trees and comparing them with the gene expression changes of more than 100,000 cells undergoing iEP conversion...tags often had similar gene expression pattern changes and reprogramming outcomes, suggesting that outcomes... expression and clonal history associated with changes in cell identity. The Morris Lab used CellTagging... processes where cells are undergoing identity changes, such as the differentiation of induced pluripotent...

...work? As illustrated in the Figure below, the “A” overhang on the blue PCR product insert comes from using...TOPO kits provide vectors or cloning arms with overhanging 3´ deoxythymidine (T) residues that are covalently...positive clones. Once you introduce your 3’-end “A” overhang insert, the magic of TOPO cloning happens. Basic...methods to ensure your insert retains the 3' A overhang: Use a mixture of proofreading enzyme and Taq...

...final plasmid by sanger sequencing. DNA replication by PCR has error rates that range from roughly 1 per...interest. You can use similar processes to add overhangs to your insert of interest for Gibson assembly...chance for mis-priming, so you can use a pretty wide range of annealing temperatures, but you may need to increase...

...the spatial and temporal arrangement of chromosomes and how these arrangements may affect nuclear processes...known for its ability to bind DNA targets without changing them. Thus, it has been widely adapted for a wide... Jennifer Tsang...

...phaffii (formerly known as pichia). What is this new-fangled yeast? It’s actually not new at all, but it’s not... Ploidy Usually diploid, but spontaneously changes Usually haploid Usually haploid Pathology...workhorse. This yeast can also give more protein bang for your media buck - it grows at higher cell densities...their peroxisomes will proliferate upon a media change into methanol and peroxisomal autophagy can be ...

...spatially limited studies and can fluctuate with changing culture conditions. The use of low intensity light...activity persisted in the inactive state. The dynamic range, defined as the difference between the lowest and... reduced system leakiness and increased dynamic range to around 100-fold. These improvements will allow...Applications of these systems The large dynamic range of these systems ensures that they can be used with...

...picked up by the sensors. You can adjust this by changing the voltage for the particular fluorophore prior... Figure 2: (A) a smeary dot plot showing a range of antibody readouts. (B) A gate on the smeary dot...histogram plot for a smeary antibody readout. The orange peis the baseline expression of the protein of ...yes-no histogram plot shown previously. Here, the orange peak is a baseline expression of a protein while...

...this, throughout the year we rolled out multiple changes to our website, including the recent homepage and... a look at the least visible part behind these changes: the backend technology that powers the new website...website search. Tackling findability from multiple angles    We assembled a cross-team team, with members....   The team approached the issue from multiple angles: the overall user experience of our website, the...

...S, Trevino AE, Scott DA, Inoue A, Matoba S, Zhang Y, Zhang F (2013) Double Nicking by RNA-Guided CRISPR...insertion) provided a template containing compatible overhangs is available. Cas9 can also be altered to generate...Ruis B, Oh S, Tomkinson AE, Hendrickson EA, Giovannangeli C, Jasin M, Brunet E (2014) Chromosomal Translocations...

...Help identify the skills you will need to make a change Provide perspective on core values at another company...coachable”. That means the mentee needs to be open to change and constructive suggestions and feedback. The ...their advice made a difference, helped you make a change for the good or resulted in a leap in your skills...These are all normal outcomes. If you have a broad range of mentors you will always have people you can turn...

...editing by a miniature CRISPR-Cas12f nuclease Quanjiang Ji’s lab describes an effective miniature Cas ... its packaging into virus-like particles. Feng Zhang’s lab adapted the native PEG10 RNA sequences to instead...et al. 2021.   Prime editing improvements By Angela Holmes Several labs have recently shared some exciting...PEDAR is capable of correcting larger genomic rearrangements as demonstrated in a mouse model of tyrosinemia...