...to the challenge. Find the PHP Plasmids Here! Their Solution: Use the CREATE method to engineer two new...3) they have limited toxicity. However, a key challenge of using AAVs for neuroscience research is the...AAV.PHP.S targets the PNS since it was actually engineered to target the CNS. How well do these AAVs transduce...takeaways: Two new AAV capsids (figure 1) were engineered with the CREATE method (figure 2) to allow for...References 1. Chan, KY, Jang MJ, Yoo BB, et al. Engineered AAVs for efficient noninvasive gene delivery ...

...biological engineering. 2010;4(1):1. Pubmed PMID: 20205762. PubMed Central PMCID: PMC2822740. 2. Engler C, Kandzia...Knight TF. Engineering BioBrick vectors from BioBrick parts. Journal of biological engineering. 2008;2:5...neighboring fragments require compatible cohesive ends Engler et al., 2008 Gibson Cloning Gibson assembly method...MXS-chaining method have used the technique to engineer various constructs with applications in flow cytometry...

...Aequorea victoria. It has a fluorescent emission wavelength in the green portion of the visible spectrum ...study of dynamic, physiological processes. Re-engineering GFP to increase its range of color and application...its sequence was elucidated, scientists began engineering new versions of GFP through mutagenesis in order...iterations of fluorophores are constantly being engineered. Table 1 lists a few common fluorescent proteins...component (cyan derivatives) Y66H Blue-shifts the wavelength (blue derivatives) Y145F Increases quantum ...at high concentrations T203Y Red-shifts the wavelength (yellow derivatives) A206K Interferes with ...

...one of those stories. Viral vectors are viruses engineered to be safely used to deliver a genetic payload...depending on their serotype, are particularly challenging. Their efficacy in any given system can be affected...significant time and removes a variable from the lengthy process of validating and using an AAV in a specific...activated, respectively, by substrates, light, or engineered ligands (Figure 2). Biosensors, which include...accessible. The supporting data and community engagement required to do this drove Addgene to develop ...partnerships, and new platforms, from our early engagement with UPenn to our launch of the redesigned Data...

...phage-assisted continuous evolution and protein engineering to generate seven new prime editors, PE6a–g, ...(2023). Phage-assisted evolution and protein engineering yield compact, efficient prime editors. Cell,...delivery and reduce off-target effects of gene therapy, Feng Zhang’s lab has developed a system called DIRECTED...local proteome, but some applications have been challenging or inaccessible. The Ting lab recently made two... increase the utility of this technique.  By engineering a light-sensitive LOV domain into the TurboID...organisms. Lee, S.Y., Cheah, J.S., et al. (2023). Engineered allostery in light-regulated LOV-Turbo enables...

...alongside some creative (and significant!) protein engineering. They combined Prime editing with an integrase...right integrase for this new approach proved a challenge. The old standby of CRE/flp didn’t work – it turns...potential applications. But they weren’t done engineering quite yet – surely, they thought, the integration...optimized the specific attachment sites via sequence engineering, enabling even more efficient integration activity...system that could integrate genes up to 36 kb in length, with an integration rate of 10-55% - the PASTE...: Adding Precision and Flexibility to CRISPR Engineering CRISPR 101: RNA Editing With Cas13  CRISPR Cheat...

...-targeting cutting as full-length gRNAs Lastly, the effects of guide length on off-target cuts was explored...17-18 bp) have less off-target cutting than full-length guides (19-20bp)?  Small-scale studies (Tsai et...explored using full-length (19-20 bp) or short (17-18 bp) guides. When comparing the toxicity of guides...short guides had reduced toxicity compared to full-length guides. Reduced toxicity of short guides could ...28474669. Pubmed Central PMCID: PMC5424143. 2. Tsai, Shengdar Q., et al. "GUIDE-seq enables genome-wide profiling...

...Nguyen NT, Zheng Z, Gonzales APW, Li Z, Peterson RT, Yeh J-RJ, Aryee MJ, Joung JK (2015) Engineered CRISPR-Cas9...-associated virus (AAV).  At 984 amino acids in length, Cas9 from Campylobacter jejuni (CjCas9) is even...first non-Cas9 CRISPR protein adapted for genome engineering was Cas12a (formerly Cpf1), a nuclease that generates...Unconstrained genome targeting with near-PAMless engineered CRISPR-Cas9 variants. Science 368:290–296 . https... genome engineering...

...the Wyss Institute for Biologically Inspired Engineering. CRISPR/Cas9 is an enormously plastic tool and...that can reassemble when exposed to specific UV wavelengths or chemicals (Polstein and Gersbach, 2015; Zetsche...activator by simply providing a new gRNA rather than engineering an entirely new protein. This makes it much cheaper...the Wyss Institute for Biologically Inspired Engineering.     Alejandro (Alex) Chavez is a member of the...Church GM (2015) Unraveling CRISPR-Cas9 genome engineering parameters via a library-on-library approach....Genome-scale transcriptional activation by an engineered CRISPR-Cas9 complex. Nature 517:583–588 . https...

...unfamiliar, scFvs are often the building blocks for engineering proteins. So let’s get acquainted with scFvs....plasmid in bacteria (Ahmed et al., 2012). One challenge of bacterial expression is the proper formation... antibodies to be used as building blocks for engineered proteins. This flexibility is largely due to ...When these molecules, called Bispecific T-cell engagers (BiTE®s), bind CD3 on T cells and a tumor-specific...minimal systemic side effects. scFvs are part of engineered chimeric antigen receptors (CARs). CARs are a...Bruccoleri RE, Haber E, Crea R (1988) Protein engineering of antibody binding sites: recovery of specific...

...mediated transduction and expression of the full-length transgene in vitro and in vivo (Allocca et al.,..., the fragments reassemble and produce the full-length gene. Below are overviews and drawbacks of four...and B of the transgene to reconstitute the full-length transgene (Chamberlain et al., 2016). It’s generally...halves of the transgene will reconstitute the full-length transgene.  Drawbacks:  Trans-splicing occurs...sequence, the 3’ half of the transgene. The full length gene is reconstituted by either the overlapping...10.1038/mt.2013.184 Lai Y, Yue Y, Liu M, Ghosh A, Engelhardt JF, Chamberlain JS, Duan D (2005) Efficient in...

...the best ways to make you and your presentations engaging and memorable. If you read my story above about... results, striking images, creative ideas, and engaging stories as you encounter or experience them. Drop...everything I allow 1 slide for every 1 minute of talk length, and then I cut a few more slides. Do not run over...to be conversational - the audience should feel engaged and not like they’re sitting through a pre-recorded...experience you can experiment with other ways to engage the audience. Take an informal poll by asking for...from the speaker, but actually makes for a more engaging session. The best speakers use no notes and can...

...all cases, the result can be quantified by the strength of the signal in comparison to a control sample...in the synthesis of DNA strands of all possible lengths because extension is terminated upon the incorporation...become obsolete. For instance, the popular genome-engineering tool, CRISPR, may one day make techniques used...new prenatal test for sickle-cell anemia." New England Journal of Medicine 307.1 (1982): 30-32. PubMed...Andrew Cho, and Ashok B. Kulkarni. "Overview: engineering transgenic constructs and mice." Current Protocols...254.5028 (1991): 59-67. PubMed PMID: 1925562. 10. Feng, Yanxiao, et al. "Nanopore-based fourth-generation...

...Precision genome editing faces the challenges above and more. One chief challenge is upregulating HDR in mature...complex oligonucleotide treatment comes with many challenges, researchers have begun to explore genome editing...necrosis compared to unedited animals. In grip strength, specific force, and other muscle tests, muscle... the time to clinical approval. Even with the challenges precision editing faces, it’s encouraging to ...Central PMCID: PMC3922311. 6. Tabebordbar M, Zhu K, Cheng JK, Chew WL, Widrick JJ, Yan WX, Maesner C, Wu EY...

...Chief Software Engineer at Ginkgo Bioworks. He also holds a SB and a Masters in Engineering from MIT, both...received his Bachelor of Science degree in Computer Engineering from Tufts University and his Master of Arts ...Computer Science.   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...

...students develop their communication skills and engage with the public. The organization runs a wide variety...picked a theme (Bio-Medley) to showcase SITN’s strength in the biomedical sciences and decided to offer...offer a varied format event to keep the audience engaged. The day kicked off with introductory lectures ...presenters and board members enabled attendees to engage in small group discussions. The afternoon program...Addgene would love to hear about your efforts to engage the public - let us know in the comments section...

...straight-forward, concise explanation of how to adapt our engineered devices, recombinase-based state machines for...of inputs, may be used to better understand and engineer cellular processes that are influenced by temporally...primer binding sites (designed to have 20-30 nt length, ~60 oC annealing temperature, and ~50% GC content...over a time series of increasing input exposure lengths and found that two hour inductions were sufficient...al. "Serine recombinases as tools for genome engineering." Methods 53.4 (2011): 372-379. PubMed PMID: ...Check Out More Lu Lab Plasmids Find More Genome Engineering Tools  ...

...bacterial infection, she shared that tweet with Jan Zheng, a UX designer that she knew. “We had actually met... met at a lindy hop swing dance event,” Zheng says (networking can happen anywhere!). The pair quickly...inception of Phage Directory ~1 year ago, Sacher and Zheng found that they could expand their role in the phage...because there is so much information out there,” Zheng says. In the past year, they found that phage information...potential for successful phage therapy, Sacher and Zheng also saw the need to archive phage reagents. If ...

... W., Li, Y., Feng, D., Ho, A., Nicholas, E., Hirokawa, K.E., Bohn, P., Joines, K.M., Peng, H., Hawrylycz...forming a neuronal loop which is activated and strengthened during memory formation and memory retrieval...modified and used as tools for tracing. With some engineering, RABV and HSV, but also others including pseudorabies...been found naturally or have been genetically engineered to have differences in their capsid proteins....S.K., Deverman, B.E., and Gradinaru, V. (2017). Engineered AAVs for efficient noninvasive gene delivery ...Koch, C., Bernard, A., Dang, C., Jones, A.R., and Zeng, H. (2014). A mesoscale connectome of the mouse ...

... Greg Lohman, a biochemistry researcher at New England Biolabs. When do you need a high fidelity ligase—and...Optimization through high-throughput profiling New England Biolabs® (NEB) researchers recently published a...possible pairing resulted in a product of a different length, with products identifiable and quantifiable by...Greg Lohman is a biochemistry researcher at New England Biolabs, specializing in the enzymology of DNA ...G., et al. "Cloning, nucleotide sequence, and engineered expression of Thermus thermophilus DNA ligase...17428930. PubMed Central PMCID: PMC1933048. 16. Cheng, Changming, et al. "Electrochemiluminescent assay...