We narrowed to 812 results for: eng

...the Wyss Institute for Biologically Inspired Engineering. CRISPR/Cas9 technology has revolutionized the...the fields of molecular biology and bioengineering, as it has facilitated the development of a simple ...-competent) Cas9 interacting with either a full-length gRNA (20nt complementarity to target site) or truncated...complementarity to target site). When complexed with a full-length gRNA, native Cas9's two catalytic domains, HNH ...different perturbations. Consider, for instance, the challenge of attempting to simultaneously and selectively...of modulating gene expression (6). Altering the length of gRNAs targeting different sites of interest ...orthogonal Cas9 species. Within a single cell, full-length guides targeting one set of genetic loci can be...

...characterizing the strength a species absorbs or reflects light or radiation at a specific wavelength, and the ...light or electromagnetic radiation at another wavelength (Heinrichs, 2009). When a fluorophore absorbs...difference in emission results in a slightly longer wavelength emitted than was absorbed. This phenomenon is...known as the Stokes Shift, or the difference in wavelength between peak excitation spectrum and emission...differences in their excitation and emission wavelengths, which are pivotal in their application in biotechnology...Hayashi-Takanaka et al., 2014). Other dyes may prove challenging to stain intracellular compartments in in vivo...applications that involve ranges of pH, and significant lengths of time, the stability of fluorescence over time...

...requirements, and photon emission wavelengths. In addition, researchers have engineered a variety of improved luciferases... and engineered to be extra bright. Other luciferases have been modified to change the wavelength of light...gracilirostris) Engineered  462 19 Furimazine None Optional AkaLuc(derived from Photinus pyralis) Engineered   677...Cofactors/substrates Required Not required Signal strength Lower Higher Sensitivity Higher Lower Background...assays Available Well-established Natural and engineered luciferases Although firefly luciferase was the...Properties of luciferase enzymes Name Natural or engineered Peak emission (nm) Size (kDa) Substrate Cofactors...substrates and measure luciferase output at different wavelengths. Addgene provides empty backbones with the luciferase...

...Cas family of proteins has many useful genome engineering enzymes, each with their own unique features ... and quirks. Here, we will review the genome engineering abilities of the lesser-known Cas enzymes while... DNA targeting Cas enzymes Cas9 – The genome engineering workhorse You rarely hear the word ‘CRISPR’ without...hearing ‘Cas9’. The Cas9 nuclease is a genome engineering powerhouse, and for good reason. It is guided...strand break. Cas9 has been studied for genome engineering purposes far more than any other Cas family protein...making Cas9 an adaptable choice for many genome engineering endeavors. For more information on Cas9 variants...curious feature of this Cas is that once it is engaged by its target, it will non-specifically cut ssDNA...

...without the challenges typically occurring with a fluorescence-based approach. These challenges can include...Addgene! Measuring messengers: growth factors Growth factors present unique challenges for study. These ...in their natural context is one of the biggest challenges in biology. From tumor suppressors to growth ... activity. Together, these limitations create challenges for measurements needed to move discoveries from...(Laissue et al., 2017). HiBiT overcomes these challenges and enables precise quantification with with ...factors that utilize exosomal transport is quite challenging. Work by Shibata et al. (2023) used HiBiT to ...endothelial growth factor A (VEGF-A) is one of these challenging proteins. VEGF-A mediates angiogenesis through...

...reprogramming  Regulation of telomere length with TZAP Maintaining telomere length is important for cellular function...on regulating telomere length with TZAP Multiplexible plant genome engineering toolkit The Voytas Lab ... Segment on PhoCl LSSmCherry1 & RDSmCherry1: Engineering and directed evolution of new RFP variants The...were developed as part of a larger strategy to engineer improved red fluorescent proteins (RFPs) and, ... of base editing. Kim et al. used natural and engineered Cas9 variants to develop five new base editors...The Denchi lab investigated an aspect of telomere length regulation called telomere trimming and have proposed...ZBTB48) in regulating the upper-bound of telomere length. Telomere trimming can be assayed by looking for...

...has been a lot of engagement with the Athena SWAN Charter and it has really strengthened the network of ...Laboratory, where she often feels more like an engineer than a biologist. Their focus at the lab is on...overcome them, and why she spends some of her time engaging with others on Twitter, among other things. MoClo...fluorescent proteins. Visualizing fluorescence can be challenging in plants because of the auto-fluorescence of...similar resource for those involved in monocot engineering projects. The MoClo tools will work in any dicot...science, and many other topics. What’s the value of engaging about science in that way for you? Patron: I started...Gate Modular Cloning Toolbox for Plants. Carola Engler, Mark Youles, Ramona Gruetzner, Tim-Martin Ehnert...

...The emitted light is at a longer wavelength than the exciting wavelength. Thus, FPs are proteins with this...by a specific excitation wavelength. This means that the emission wavelength can change. In a few cases...FPs classified by the emission color (emission wavelength range) FPs are usually classified by emission...emission color as outlined below (or emission wavelenght range).  By mutating GFP, the variants blue FP (BFP...Infra-Red ≥ 670 nm In addition to emission wavelength range, there are other traits that need to be...named after George G. Stokes) is the shift in wavelength from excitation to emission. For most FPs, Stokes...sure their emission light does not overlap in wavelength. In many microscopes the filters are not narrow...

...Open Yeast Engineering Project (OYEP). Through this project, community members will engineer yeast to glow...that anyone can engineer yeast to do similar things. The focus is to make yeast engineering accessible to...are skilled with audiovisual media, software engineering, business consulting, mathematics, and more. ...and in his free time, he is pursuing a plant engineering project at BosLab. Once he optimizes plant cell...knowledge with the community by teaching a DIY plant engineering workshop. Community projects at BosLab Not all...became a member at BosLab because she wants to engineer microbes. We’ve ordered a few yeast plasmids from...diversity in different soil conditions/locations but to engage the local community (including local garden and...

...BioBuilderClub BioBuilderClub engages high school teams around the world who combine engineering approaches and scientific...are asking. Teachers are also asking, “How do I engage my students in real-world lab activities? I’m trying... Kuldell from MIT’s Department of Biological Engineering. Built primarily from her 20 years of college...classroom are characterized by intense student engagement and joy, rather than a perfect lesson plan or... boost interest and understanding around the engineering of biology. Participating schools are provided...BioBuilder inspires students and teachers to apply engineering principles to biological systems. Today, what...BioBuilder module allows students to explore how engineering fields rely on rapid prototyping to develop and...

...nuclease domain. Feng Zhang's group utilized the bi-lobed architecture of Cas9 to engineer a series of "split...memory One of the goals of synthetic biology is to engineer cells capable of recording permanent "memories...display and high throughput binding assays, to engineer photoswitchable dimers which show over a 50 fold...activating endogenous genes Moritoshi Sato's Lab has engineered CRISPR-Cas9 to create a photoactivatable transcription... or (4) multiple sgRNAs with Cas9 for genome engineering. The versatility and ease of generating complex...potential applications of CRISPR/Cas9 in genome-engineering. One particularly attractive application involves...genes involved in neurogenesis, which had proven challenging using the existing dCas9-VP64 activator. The ...

...editing (Feng Zhang Lab 2015)  Fluorescent Proteins Plasmids and Resources Reference Feng, S., Sekine...producing fluorescent signal. Image modified from Feng et al. 2017 under CC-BY license. Many split-FP...conditions. Shyu et al. Biotechniques. 2006 Chang-Deng Hu Green sfGFP Split super-folder GFP, our...fluorescent proteins for endogenous protein labeling. Feng et al. Nat Commun. 2017 Bo Huang mNeonGreen3 ...conditions. Shyu et al. Biotechniques. 2006 Chang-Deng Hu Venus Improved N-terminal fragment VN155(...signal-to-noise ratio. Kodama et al. Biotechniques. 2010 Chang-Deng Hu mVenus pET-BiFC contains both fragments of...fluorescent proteins for endogenous protein labeling. Feng et al. Nat Commun. 2017 Bo Huang sfCherry3 Improved...

...Addgene to advance their research. Louis Bengyella Dr. Bengyella’s work has focused on elucidating the molecular...our first two plasmid grant awardees: Drs Louis Bengyella and Kwabena O. Duedu, both from the University...in Ghana. Read on to learn more about doctors Bengyella and Duedu and how they plan to use plasmids from... microbial pathogens. This work has given Dr. Bengwella expertise in using bioinformatic and proteomic...associated with pathogenesis. In his future work. Dr. Bengyella would like to use the expertise he’s developed...development of uterine tumors. According to Dr. Bengyella, this class of tumors is “a major public health... Ghana. We’re excited to see what impact Dr. Bengyella’s work will have on women’s health in Ghana and...

...funding, bioethics in the age of CRISPR, and the challenges of science becoming increasingly politicized...., “The Professional Scientist,” addresses the challenges of succeeding in science if you’re anything other... course the rapidly changing world of genome engineering via CRISPR.  Though the individual topics are...ramifications. Readers unfamiliar with academia’s challenges with publication and funding may be dismayed ...The three chapters in this section describe the challenges of trusting science in a “fake news and post-...has played in these fields, and seeing names like Feng Zhang, Jennifer Doudna, Karl Deisseroth, and Ed ...describes the author’s thoughts on science’s biggest challenges over the coming years. It is arguably the most...

...editing efficiency, and target access were all challenges for these early base editors. Further innovations...tackle this problem, Benjamin Kleinstiver’s lab engineered new variants of S. pyogenes Cas9. While the wild-type...sequence of NGG (where N represents any base), their engineered variants are more permissive: SpG can target ...Kleinstiver lab also worked on the opposite project: engineering Cas9 variants with a preference for specific ... L. A., & Zhang, F. (2013). Multiplex genome engineering using CRISPR/Cas systems. Science (New York, ...Gene Editing to Treat a Rare Genetic Disease. New England Journal of Medicine, 392(22), 2235–2243. https:... A., Thuronyi, B. W., Wilson, C., Koblan, L. W., Zeng, J., Bauer, D. E., Doudna, J. A., & Liu, D. R. (...

... in a variety of cells and organisms. However, lengthy genetic payloads and insufficient transcription...assembled tRNA-gRNAs is placed downstream of a lengthy promoter, which is used to drive the expression...efficient MBE with such a strategy, the need for lengthy promoters, such as U6 or EF1α promoters, has limited...derived from human or plant source), by tuning the length of 5’ leader sequences of a given tRNA to achieve...either from human or plant sources) in both the strength as a promoter and the activity to release gRNAs...results of their MBE efforts, the Gao Lab also engineered a DAP array for prime editing, achieving up to...also enable more convenient multiplex genome engineering of crops or allow for sophisticated CRISPR screenings...

...photobiology: why use light? In synthetic biology and bioengineering, scientists seek to directly probe and build...problems; inexpensive tools such as LEDs can be engineered to deliver precise, consistent, and controllable.... Complicated patterns of light intensity and wavelength can be used to study dynamic processes. Conversely...Conversely, varied input patterns may also be used to engineer new pathways with sophisticated gene expression... of edge detection. Tabor et al. subsequently engineered a second photosensitive TCS activated by green...-friendly. They experimented with the promoter strength of various components, removing inducible promoters...could be made suitable for various scientific or engineering applications.   References Refactoring and optimization...

...Cas9 and Cas12a, creates challenges for in vivo delivery and cell engineering applications. CasMINI, a...gene perturbation The Penn State Protein Ladder Engineered miniature CRISPR-Cas system for mammalian genome...flexible, modular tool for labeling. Here, Liu et al. engineer a chimeric single chain variable fragment (scFv...gels and can be detected on Western blots via engineered immunoglobulin binding domains.  Santilli, R.T.... https://doi.org/10.1038/s41598-021-96051-x Engineered miniature CRISPR-Cas system for mammalian genome... Figure 4: Overview of CasMINI for genome engineering applications. Image from Xiaoshu, et. al. 2021...system opens up new possibilities in both cell engineering and gene therapy.  Xiaoshu X, et al. Molecular...

...technology since its initial applications for genome engineering in 2012.  Many of these advances have stemmed... of dCas9; and 2) CRY2FL-VP64, a fusion of full-length CRY2 and the transcriptional activator domain VP64...PAM and had similar targeting specificity as full-length Cas9 (flCas9). When triggered by blue light (470nm...light-dependent (Yu et al., 2020). To express the N-Cas9, Haifeng Ye’s lab used the bacterial phytochrome BphS which...employed a photoactive strategy which had been engineered from naturally occurring photoactive proteins... specific site on the Cas9, the group used an engineered pyrrolysyl tRNA/tRNA synthetase pair which would... Grimm D, Correia BE, Eils R, Niopek D (2018) Engineered anti-CRISPR proteins for optogenetic control ...

...has developed a miniature Cas system (CasMINI) engineered from the uncultivated archaea Cas12f (Un1Cas12f1... system. The CasMINI system was developed by engineering the Cas12f scaffold gRNA and making strategic...CRISPR editing with a hypercompact Cas12f1 and engineered guide RNAs Yong-Sam Kim’s lab also created a ...the gRNA scaffold through five rounds of gRNA engineering.  They did not modify the Cas12f1 enzyme. The...facilitate its packaging into virus-like particles. Feng Zhang’s lab adapted the native PEG10 RNA sequences...plasmid tools to optimize prime editing systems. Engineered pegRNAs improve prime editing efficiency In the...the 3’ end with a structured RNA motif. These engineered pegRNAs (epegRNAs) protect the RNA from degradation...