... been modified for genome engineering . Prior to CRISPR, genome engineering approaches like zinc finger...rapidly become the most popular genome engineering approach. Engineered CRISPR systems contain two components...Cas-mediated genome engineering. 2013. Wang H, Yang H, Shivalila CS, Dawlaty MM, Cheng AW, Zhang F, Jaenisch...Activation or Repression Epigenetics Multiplex Genome Engineering Genome-wide Screening What are pooled libraries...mutation within the targeted gene. However, the strength of the knockout phenotype for a given mutant cell...generation of high fidelity enzymes have been engineered using cell-based selection approaches. Yeast-...target (termed left & right homology arms .) The length of each homology arm is dependent on the size of...

...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...

...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 ...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...Santa Cruz, where her research is focused on engineering nanoparticles as drug delivery systems. She is..., Raj, et al. "Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach....

...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 ...

...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...

...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...

... 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...

...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...

...while writing each section. Research challenge The Research Challenge motivates the request for application...creates strong, well-directed meaning and delivers strength to the sentence. Below are the typical sections...how you, the expert, will address the research challenge. Now is the time to make bold statements. While...solution is the best approach to the research challenge. Demonstrating expertise in the field and past...how your innovation will address a variety of challenges and impact the field - its encouraged to dream...time budget writer, you may find this section challenging because it’s hard to determine the exact value... differs from publication writing and may be challenging to master at first. Additionally, reach out to...

...proteins or TALE proteins, proteins had to be re-engineered for each gene, making wide-scale gene activation...gene activation: rather than requiring protein engineering for each loci, CRISPR/Cas systems only require...methods rely on fusion proteins while others re-engineer components of Cas systems themselves. SunTag, ...Mediator (SAM) Description SAM uses specially engineered sgRNAs to increase transcription. This is done... through creating a dCas9/VP64 fusion protein engineered with aptamers that bind to MS2 proteins. These...activation of bacterial gene expression using an engineered CRISPR-Cas system. Nucleic Acids Research 41:...Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature 517:583–588 . https...

...structure-guided engineering of the PAM-interacting domain of Cas9. After multiple rounds of engineering, they developed...the "perfect" guide sequence for your genome engineering experiment. With CRISPR entering the clinic for... Nureki lab generated SpCas9-NG, a rationally engineered Cas9 with NG PAM compatibility, as the name implies...R.A. et al. PAM-flexible genome editing with an engineered chimeric Cas9. Nat Commun 14, 6175 (2023). DOI...Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants. Science 368,290-296(2020...Plasmids available here)   Nishimasu, H., et al.,Engineered CRISPR-Cas9 nuclease with expanded targeting ...Requirement and Expanding Beyond Cas9 xCas9: Engineering a CRISPR Variant with PAM Flexibility History...

... Kucera, M.Sc. and Eric Cantor, Ph.D. from New England Biolabs. Golden gate assembly limitations Embraced... assemblies that enable biosynthetic pathway engineering and optimization. However, current best practices... reduced these limitations through the use of engineered Type IIS restriction enzymes and careful choice...redesign was guided by both advances in the re-engineering of the original BsaI-HF® Type IIS restriction...In conjunction with BsaI-HFv2 (NEB #R3733), re-engineered to provide improved Golden Gate performance, ... Kucera, M.Sc. and Eric Cantor, Ph.D. from New England Biolabs. Becky Kucera is a Principal Development...Development focusing on DNA cloning and assembly, New England Biolabs, Inc.         Eric Cantor is a molecular...

...recombinant protein purification and metabolic engineering. If instead you want to build genetic circuits...biology MoClo applications, but there are more! Engineering disease resistance through gene stacking, increasing...localization signals, plant markers, and reporters (Engler et al., 2014). This kit is a great starter for ...vectors, several delivery systems which can be engineered into the TUs, and even viral gene silencing vectors...kit has a focus on transformation and genome engineering. This is a specialized system with parts including...confined to that arena! The Mammalian Toolkit was engineered specifically for the types of expression and ...Multiplex MoClo Toolkit for Extensive and Flexible Engineering of Saccharomyces Cerevisiae.” ACS Synthetic Biology...

...excitation wavelength (Salomonnson et al. 2012). They can also be excited efficiently at wavelengths between...development of cell sorters together with the engineering of fluorescently labelled antibodies made it ... lymph nodes at different times after antigen challenge, scientists were able to track where GFP labelled...et al. 2016, Cavanagh et al. 2008)? Technical challenges of two photon microscopy Intravital imaging is... at steady state or under physio-pathological challenge. However technical specificities have to be taken... also important to finely tune the excitation wavelength as it has been shown that even a small incremental...incremental change of the excitation wavelength can significantly affect emission intensities from fluorescent...

...Ellis does research in synthetic biology and bioengineering including a recent effort to construct a synthetic... SynBio. Chapter 3: Synthetic biology as the engineering of biology at the level of DNA using modular,...in the Ellis lab including yeast and E. coli engineering, microbial cooperation, and antibiotic production...lectures synthetic biology for the Department of Bioengineering. He graduated from Oxford University and then...supervision of Prof Jim Collins and specialising in engineering gene regulation in yeast. Dr. Ellis is now one...undergraduate projects. Special thanks to our sound engineer/producer, Eddy Page! Edward B. Page is a Digital...Production Specialist at Addgene. He is the audio engineer and producer for the Addgene Podcast.      Additional...

...of optogenetics integrates optics and genetic engineering to measure and manipulate cells (frequently neurons...red dots). When exposed to light of the correct wavelength, the pore opens, cations flow into the cell (...channels or pumps that absorb light at specific wavelengths. Upon activation by light, these channels and...occurring microbial opsins that respond to different wavelengths of light, like blue or yellow light. These various...proton efflux. Researchers have used genetic engineering to improve these natural opsins - by inducing...the neuron. NpHR Variants. Variants have been engineered with enhancements such as: Human codon optimization...blue-green light. Mac Variants. Mac variants have been engineered to include enhancements such as: Improved photocurrent...