...PMC4923196. Panchapakesan, Shanker Shyam S., et al. "Ribonucleoprotein purification and characterization using...RNA is not inherently fluorescent and can be challenging to track intracellularly, but aptamers permit...that bind metabolites, leading to conformational changes that control the expression of coding RNA. Spinach-based... bind DFHO and fluoresce in the red and orange channels, respectively. Both Red Broccoli and Orange Broccoli...orange (TO) and fluoresces in the orange or red channel, depending on the structure of the TO derivative... used Mango to study RNA-protein complexes (Panchapakesan et al., 2017) and short non-coding RNAs (Autour...Hauer, and Diane E. Griffin. "Development and characterization of sindbis virus with encoded fluorescent ...

... Hirsch ML, Li C, Bellon I, Yin C, Chavala S, Pryadkina M, Richard I, Samulski RJ (2013) Oversized AAV...transgene to reconstitute the full-length transgene (Chamberlain et al., 2016). It’s generally accepted that some...occurs via this region of overlap, but the exact mechanism is not yet known. Drawbacks:  Generally an ... either the overlapping or the trans-splicing mechanism. The splice site donor and acceptor sequences ... . https://doi.org/10.3389/fnins.2017.00503  Chamberlain K, Riyad JM, Weber T (2016) Expressing Transgenes....2015.140 Frischmeyer PA (2002) An mRNA Surveillance Mechanism That Eliminates Transcripts Lacking Termination...Lai Y, Yue Y, Liu M, Ghosh A, Engelhardt JF, Chamberlain JS, Duan D (2005) Efficient in vivo gene expression...

...generate the strictly monomeric EGFP variant, mEGFP (Zacharias et al., 2002). GFP needs oxygen Chromophore formation...incorporation into proteins are steps in that direction (Chatterjee et al., 2013; Hilaire et al., 2017). GFP can ...specific RNA stem loop-structure (Querido and Chartrand, 2008). Finally, a nuclease-deficient Cas9 can...post-translational modification with fatty acyl chains. For instance, myristoylation requires an N-terminal...microscopy (University of Amsterdam). He develops, characterizes, and uses genetically encoded fluorescent probes...on twitter: @joachimgoedhart.   References 1. Zacharias, David A., et al. "Partitioning of lipid-modified...of Green Fluorescent Protein Fused with a Single Chain Fv Protein." Journal of Biological Chemistry (2000...

...to a negatively charged residue, aspartate or glutamate, imparting the negative charge associated with...carry only a single negative charge as opposed to a double negative charge in the phosphate group. Additionally... acids fail to recapitulate the true steric and charge-based nature of the phosphoryl-modification; they...phosphorylated intermediate of a protein for characterization. However, phosphoproteins are often low abundance...incorporate phosphoserine A Sep-tRNA synthetase, which charges this tRNA with phosphoserine A specialized elongation...an optimized elongation factor that accepts Sep-charged tRNAs during translation. Many ‘flavors’ of Sep-OTS...et al. "RNA-dependent cysteine biosynthesis in archaea." Science 307.5717 (2005): 1969-1972. PubMed PMID...

...construct expressing channelrhodopsin 2 (ChR2), a blue light sensitive cation channel, fused to mCherry ...emitting diodes, thermal sensors, and microfluidic channels. These features enable the fibers to emit light...stimulation of its associated fibers across multiple channels and can also record tissue temperature proximal...considerations of actually using it - the device has a rechargeable battery, is relatively small, and can be detached...Sands, J., Sahasrabudhe, A., Anikeeva, P., & Chandrakasan, A. P. (2020). Modular Optoelectronic System...Garwood, I. C., Yan, W., Niroui, F., Fink, Y., Chandrakasan, A., … Anikeeva, P. (2023). Multifunctional ...Addgene blog Plasmid tools for microbiome studies Channel rhodopsins for optogenetics Viral Vectors 101: ...

...isotype? Antibodies are composed of light chains and heavy chains made up of variable regions and constant...regions, just the heavy chain constant regions or solely the Fc region may be changed.   Chimeric antibodies...antibodies can help A chimeric antibody is made by changing an antibody’s natural isotype to that of a different...will react with the same secondary. If you were to change one of the antibody’s isotypes, however, you could...be converted? Isotype conversion is a unique characteristic of recombinant antibodies. Recombinant antibodies...containing all constant regions, just the heavy chain constant regions or solely the Fc region of a different...

...Chu H, Chan JF-W, Yuen TT-T, Shuai H, Yuan S, Wang Y, Hu B, Yip CC-Y, Tsang JO-L, Huang X, Chai Y, Yang...H-W, Cai J-P, Chan W-M, Ip JD, Chu AW-H, Zhou J, Lung DC, Kok K-H, To KK-W, Tsang OT-Y, Chan K-H, Yuen K-Y...in a publication by Xiuyuan Ou, et al. that characterized the spike (S) protein of SARS-CoV-2 (Ou et al...Jackson Laboratory. Originally developed and characterized by Stanley Perlman, of the Carver College of...for the advancement of scientific research, and mechanisms to maintain and grow these critical organizations...Chen J, Hu K, Jin Q, Wang J, Qian Z (2020) Characterization of spike glycoprotein of SARS-CoV-2 on virus...

...cells. These changes were larger than previously reported for other organisms, and the Gottschalk group hypothesizes...screens, which enables researchers to maximize the chance of transducing a only a single gRNA per cell. The... Genetically encoded voltage indicators (GEVIs) change fluorescence in real time when neurons or muscle...technical difficulties such as small fluorescence changes or a requirement for very high excitation light...these indicators difficult to work with.  The Gottschalk lab has surveyed a range of microbial rhodopsin... that can be used to monitor real time voltage changes in worms and have made these constructs available...detect action potentials in worms with fluorescence changes of up to 128% per 100 mV in the pharynx, and 78%...

...sufficient to prevent any pH change, as we still observed a slight change in pH (from 7.4 to 6.8). It’...tubes (Fig. 1). It’s important to note that the change in pH only occurred when the solution thawed, as...samples were frozen. This was clear by the gradual change in pH visible at the sample interface upon thaw...found that 30 seconds of venting attenuated the pH change caused by storage on dry ice (Fig. 3). However,...CO2 to leave the tubes, and would prevent the pH change upon thaw (Fig.4). To test this, we froze the tubes...stability (Murphy et al., 2013). However, the pH changes in our solutions are transient and return to normal... have no evidence to suggest that the transient change in pH affects AAV function. Furthermore, another...

...fit for me, then I didn’t need to explain why I changed jobs. I highly recommend taking on a similar position...become bored because I was not being mentally challenged. For these and other reasons, I started  looking...this company for several years, I was ready for a change.  Working abroad: Research in Australia I then ...for a 6 month project to validate some genomic changes, so this arrangement was ideal for everyone.  The...combination of genetic mutations. This is a particular challenge in pancreatic cancer where one drug does not work... fit in with the company and that my ideas and changes were welcomed, which is a benefit over larger companies...who are grateful for some help.  I also get the chance to contribute content to the Addgene’s Website ...

...goals can be a real challenge. On top of that, in the midst of a rapidly changing, globalized environment...training in a globalized world where individuals can exchange ideas, network, collaborate, and innovate internationally...technical and logistic support to increase the chances of success, but also nurture a supportive and protective...mentees with a broader, first-hand vision of the challenges and career opportunities that truly exist outside...international mentor creates a closer and more approachable relationship for the mentee, where (s)he feels...was much more than that. I discovered that the challenges that I am facing in my career are common problems... that many others have faced yet, still, this challenge did not hinder them from advancing their careers...

...deaminase. Base changes from C•G-to-T•A are mediated by cytosine base editors (CBEs) and base changes from A•T-to-G...capabilities. Base editors make specific DNA base changes and consist of a catalytically impaired Cas protein...target site, and the deaminase induces the DNA base change without a DNA double-strand break. But base editors...is that the editing domain must induce the base change before the Cas domain lets go of the DNA. ABEs ...hold on” long enough to the DNA to induce a base change. Because of this, ABEs are only weakly compatible... of the fetal hemoglobin gene, ABE8e made both changes simultaneously with 54% efficiency, about 7 times... DNA base swaps, so called point mutations that change C to T, G to A, A to G, and T to C, represent about...

...set-up has been changed (for instance after replacing a broken lightbulb or exchanging excitation filters...fluorescent proteins may display reversible intensity changes (Shaner et al, 2008; Bindels et al, 2017) and photoswitching...fluorescence signal, showing no or little deterioration or change of the signal during the course of the experiment...and by comparing how the fluorescence intensity changes over time, the photostability of the fluorescent...comparable (lower panel of figure 1). To avoid changes in excitation power, it is important to perform...filters). This avoids substantial changes in excitation power and will help to keep the photostability of...microscopy (University of Amsterdam). He develops, characterizes and uses genetically encoded fluorescent probes...

...McClinntock who first discovered transposons due to changes in coloration of corn kernels. Aside from corn,... or a reverse transcriptase, depending on the mechanism of transposition.  Target site: Different transposons...elements are split into two classes based on their mechanism of transposition: Class I TEs, also called retrotransposons...retrotransposons. They transpose by a ‘copy-paste’ mechanism (Fig. 1). They first copy themselves as RNA transcripts...like retroviruses both in their structure and mechanism of replication. They contain two genes: gag and...Retrotransposons are mobilized by a ‘copy-paste’ mechanism. First they copy themselves as RNA transcripts...transposons have a non-replicative “cut and paste” mechanism of transposition: they first excise themselves...

... ́-phosphate to the ligation). The structural/mechanistic reason for this differential is not known for... E., et al. "DNA ligases: structure, reaction mechanism, and function." Chemical reviews 106.2 (2006):...PMID: 2753355. 12. Barany, Francis. "The ligase chain reaction in a PCR world." PCR Methods Appl 1.1 (...PMID: 1842922. 13. Wiedmann, M., et al. "Ligase chain reaction (LCR)-overview and applications." PCR Methods... PubMed Central PMCID: PMC1933048. 16. Cheng, Changming, et al. "Electrochemiluminescent assay for detection...detection reaction/CGE method using a LIF dual‐channel detection system for direct identification of allelic...reaction-fluorescent microsphere assay for characterization of resistance-mediating polymorphisms in African...

...engineering. Chapter 1: Early research on CRISPR and what it's like to work in a competitive field. Chapter 2: ...Institute of Singapore. Chapter 3: Biology research and resources in Singapore. Chapter 4: Life as a new primary...project ideas, applying for grants, and mentoring. Chapter 5: Wei Leong Chew's lab management style and encouraging...encouraging mentees to pursue cool projects. Chapter 6: Projects in Wei Leong Chew's lab. Genome engineering...engineering, gene delivery methods, AAVs, and more. Chapter 7/Outro: Advice for researchers pursuing careers...

... the common cytokine receptor gamma chain (c) and is characterized by a lack of immune cell development... cargo (DNA or RNA) to target cells has been challenging. A variety of vector delivery technologies have...repair via homology directed repair (HDR). This characteristic makes AAV vectors especially amenable to correcting...caused by mutations in the gene RPE65. It is characterized by childhood onset of blindness and was thought...treatment appears to be both safe and effective. Challenges for AAV gene therapy Although AAV-based gene ...promising, scientists are working to overcome key challenges to their efficacy:  Pre-existing immunity: Many...10.11 (2003): 964-976. PubMed PMID: 12756417. 5. Chan, Ken Y., et al. "Engineered AAVs for efficient noninvasive...

...again and possibly mediate HDR in a “second chance” mechanism. To prevent new editing post-HDR, repair templates...also called Cas12a. In 2015, Feng Zhang’s lab characterized two Cpf1 nucleases, distant cousins of well-...Until recently, two major elements of Cpf1 characterization were missing: on-target and off-target editing...efficiency. Kim et al. and Kleinstiver et al. characterized genome-wide editing efficiency of two Cpf1 orthologs...don't know about Cpf1 In their original paper characterizing Cpf1, Zetsche et al. suggested that Cpf1 may...experimentally confirmed. Researchers have also not yet characterized the potential “sticky end” transgene insertion...Shah SA, Saunders SJ, Barrangou R, Brouns SJ, Charpentier E, Haft DH, Horvath P, Moineau S, Mojica FJ, ...

...of a messenger RNA transcript. The purpose and mechanism of polyadenylation vary across cell types, but...Prokaryotic termination   Prokaryotic termination mechanisms fall under two general categories: rho-dependent...transcriptional read-through. Chris Voigt’s lab has characterized a set of prokaryotic terminators and deposited... tails to certain RNAs. Unlike the eukaryotic mechanism which requires a consensus sequence for the addition...and may additionally act as a quality control mechanism to rid the cell of mis-folded RNAs. Eukaryotic...dependent, Polymerases I and II employ different mechanisms to terminate transcription. Polymerase I uses...process similar to the prokaryotic Rho-dependent mechanism, whereas Polymerase II termination is more complex...

...interview with Crystal Shih Podcast breakdown Intro Chapter 1: Who is Crystal Shih? Crystal's educational background... background and position at Novartis. Chapter 2: What's a lab group in industry? The collaborative nature...industry and how delegating can empower people Chapter 3: How Crystal advanced at Novartis, and how to...development conversation with your manager early on. Chapter 4: Crystal's approach to management and the importance...and being receptive to those who report to you. Chapter 5: The types of science that Crystal works on at... big picture and not just the next experiment. Chapter 6: Advice to scientists who are planning on working...Mobility between different positions at Novartis. Chapter 7: How Crystal developed her science communication...