...Emily P. Bentley in December 2024. References and Resources References Anzalone, A. V., Gao, X. D., Podracky...(Anzalone et al., 2019). Plus, it can combine different types of edits with one another. All of this is... An overview of prime editing. Created with BioRender.com. How does it work? First, an engineered ...precise than base editing (in certain circumstances) Current base editors can perform any base-to-base conversion...indels than prime editing. But for positions that aren’t well positioned within the editing window, prime...type, and goal of the experiment can all impose different constraints and affect how you select and design...5.1-fold (though up to 45-fold) higher across different genomic sites compared to PE1 (Anzalone et al....

...disease postnatally. Now, three papers published concurrently in Science have shown CRISPR can treat a genetic... Even low-frequency editing could make a huge difference in DMD, and it’s estimated that exon skipping...respectively. Although each study used slightly different methods, they each observed the same positive ...necrosis compared to unedited animals. In grip strength, specific force, and other muscle tests, muscle...disease type C. Although the three DMD studies referenced here represent a great step forward for CRISPR...many distinct CRISPR therapies to match these different mutations. For each therapy, effectiveness and...treatments may also be closer than we might expect.   References 1. Kaiser J. CRISPR helps heal mice with muscular...

...research coming out of the lab.  References and resources References Hao, Y., Yan, J., Fraser, C., Jiang...affinity. This extended-open state is key to strengthening ligand binding, mediating cell adhesion and ...then used these antibodies to show the binding preferences of the αV subunit and its β partners. Of interest...antibodies and affinity reagents to study the difference in cell morphology changes induced by integrins...antibodies comes from IPI’s work with the Boston Children’s Hospital and Harvard-based lab of Tim Springer...discovered by yeast display reveal αV subunit pairing preferences with β subunits (p. 2024.01.26.577394). bioRxiv...

... you need. Most often, early career scientists aren’t trained to have these types of negotiations. They...all of the major equipment that you use in your current lab first. Generally, major equipment can be defined..., glassware, etc), get a simple list from your current P.I., lab manager, or administrator containing ... Do online searches, and seek advice from your current administrative services, managers, and techs’ financial... is where you can start to identify which items aren’t critical to your research success. If you can get...can still go horribly awry when several factors aren’t taken into consideration. 1. Always focus on the...for the success of your science. If the resources aren’t available, then you’re entering a win-lose situation...

... for in the future! References Bubeck F, Hoffmann MD, Harteveld Z, Aschenbrenner S, Bietz A, Waldhauer...of Cas9 activity.  The field of optogenetics is renowned for enabling precise temporal and spatial control...CRISPR gene activation system, but settled on a different optimal fusion protein combination. The optimized... C-terminal (residues 714-1368, C714) halves, rendering the Cas9 non-functional when split but regaining...absence of light, Cas9 is bound by AcrIIA4 and rendered inactive. In the presence of blue light, AcrIIA4...Alexander Deiters’ lab, on the other hand, took a different approach. These researchers used a genetically...photoactivatable transcription systems to induce neuronal differentiation. Nat Methods 14:963–966 . https://doi.org/10.1038...

...However, many genes have different functions in different cell types, at different times, and in combination...T cell responses to cancer.     References and resources References LaFleur, M. W., Lemmen, A. M., Streeter...combination with different genes. Having CRISPR systems for assessing genes in defined contexts would be quite...

... tissues are nearly transparent. As Verkhusha explained it to me, the transparency window of mammalian...tumor. They wanted to find something better. Transparency window That’s exactly what they reported doing...corals, but all of them fluoresce outside the transparency window of mammalian tissues,” Verkhusha said...near-infrared fluorescent proteins within this transparency window, so now, we could see deeper.” Adding...of the time, the answers will be there.     References: Bright and stable near-infrared fluorescent ...

...meaningful changes in the lab could make a big difference in our global impact as scientists. Many scientists... boxes Some vendors take back their expanded polystyrene (a.k.a. Styrofoam or cooler boxes). We get these...receiving temperature-controlled reagents. Expanded polystyrene will likely end up in a landfill, but at least...with CO2 emissions. NEB takes back expanded polystyrene boxes for free (in the U.S.). Their cooler boxes... 1976! Millipore Sigma takes back expanded polystyrene boxes for free (in the U.S.). After you receive...Sustainable Practices to the Lab: Easy Action Items References Geyer, Roland, Jenna R. Jambeck, and Kara Lavender...

...Swanson a structural biologist and protein engineer currently working at Sanofi Pharmaceuticals in Cambridge... Rare Disorders (NORD). NORD raises money and awareness for rare disease and helps patients find the correct...first met AJ and his family, he did not seem much different than my second son of similar age. When he came...play with my kids like any other youngster. His parents, Kristi and Rico, had to keep him from running ...an organization dedicated to raising funds and awareness of FOP. Please consider running with us or making...is a structural biologist and protein engineer currently working at Sanofi Pharmaceuticals in Cambridge...

...specific compounds, links to publication references, and cross-references with cell line catalogs and collections...be adjusted based on query, reference, and shared allele search preferences to help you identify proximal...entry.) You can look up the cell lines you are currently or planning to use for helpful information on ...to learn more, check out the Cellosaurus FAQ! References Lorsch JR, Collins FS, Lippincott-Schwartz J....

...the discoveries yet to come.      References and resources References Arbabi-Ghahroudi M (2017) Camelid...Camelids. The declaration is intended to raise awareness of the economic and cultural importance of these...become appealing candidates for the detection of different cancers (Jin, et al. 2023).  So where do all these...mouse-derived single-domain antibodies. Image created in BioRender and adapted from Xu, et al., 2021.   The single-domain...antibody field has come a long way since the serendipitous discovery of heavy-chain only antibodies in ...

...for contributing their expertise! References and resources References Aschauer, D. F., Kreuz, S., & Rumpel... 1: Diagram of AAV components. Created with BioRender.com. AAV vectors have both distinctive capsids...multiple coreceptors, which may be present on different cell types and sometimes vary between target species...service.  In summary Your choice of AAV vector inherently depends on your experimental system. As we cannot... al., 2021 In addition to tissue tropism, different AAV serotypes can be useful to evade neutralizing...Transfer by Adeno-Associated Virus Vectors via Different Routes. Diabetes, 55(4), 875–884. https://doi....

... the transcript. Different Cas orthologues and subtypes function somewhat differently and can be more ...Gram-positive bacteria Jason Peters, Oren Rosenberg, and Carol Gross used two different genetic systems, targeting...Transcriptional repression (CRISPRi) in bacteria As RNA interference does not function in bacteria, most efforts ...Clostridium difficile, could be targeted and preferentially depleted from gut. While still far from the...the BE3 that further improves editing rates. References Bikard D, Euler CW, Jiang W, Nussenzweig PM, ...inMycobacterium tuberculosisusing an efficient CRISPR interference system. Nucleic Acids Res 44:e143–e143 . https...

...Emily P. Bentley in February 2025. References and Resources References Chen, L., Zhu, B., Ru, G., Meng, ...component of the nucleoside inosine. Created with BioRender.com. How does base editing work? Base transition... UGI = uracil DNA glycosylase. Created with BioRender.com. The BE3 system increased editing frequency...APOBEC3G displayed less predictable sequence preferences.  The Liu lab also used natural and engineered...yielded an editor with more flexible sequence preferences, evoAPOBEC1-BE4max, which is significantly more...the development of prime editing, which uses a different mechanism than base editing, has allowed scientists.... A dual-deaminase CRISPR base editor enables concurrent adenine and cytosine editing. Nature Biotechnology...

...redesigning viral packaging plasmids, investigating different viral production strategies or developing novel...silver stain approaches we show that there is no difference in purity or infectivity between the untreated...GFP-expressing AAV particles we did not observe any difference between untreated and sucrose treated cells. ...it will be exciting to test this approach with different capsids. Most importantly, because our AAV yields...spending less time and resources resupplying our current inventory and more time providing new materials...materials to the research community. References 1. Coroadinha, A. S., et al. "Effect of osmotic pressure on...

...sensors offer different levels of apparent dopamine affinity, with 113050 offering high apparent affinity,... various AAV serotypes can be used to achieve different patterns of expression, researchers often need...affinity, and 113049 offering medium apparent affinity. Plasmid Serotype Name 113050 AAV9, AAVrg pAAV-hSyn-GRAB_DA1h...Glutamate sensors iGluSnFR is a useful tool from Loren Looger’s lab at Janelia for neuroscientists to measure...

...work demonstrates that different repair template strategies activate different HDR sub-pathways like synthesis-dependent...live updates! My most popular tweet from the conference is shown above. The blog post I mentioned, John...post-CRISPR cleavage When I first looked at the conference agenda, I was surprised to see that it would ...Having had time to digest my notes, since the conference, I think I may have finally grasped just how ...Subscribe to CRISPR updates on our blog to keep current with the latest and greatest! Addgene resources...

...National Institutes of Health.     References and Resources References Bradbury A, Plückthun A (2015) Reproducibility...that secrete antibodies in culture and provide renewable reagents with reliable and reproducible production...made available through our usual service (see our current antibody plasmid collection) and we plan to introduce...working with the community to create a fully transparent resource where researchers can easily share, ... Trimmer JS, Görnemann J, Glanville J, Wolf P, Frenzel A, Wong J, Koh XY, Eng H-Y, Lane D, Lefranc M-P...

...specific problem. Every species is so different – an awake monkey is different from a cell in a dish. I always...want to activate two sets of neurons and see how different parts of the brain or the cells within them work...Boyden: I think because neuroscience has so many different goals: to understand how a mouse changes emotions...find ways to make these tools more selective for different ions. Right now, we can modulate electricity. ... click the link in the Addgene Alerts box. References: Independent Optical Excitation of Distinct Neural...

...This post was contributed by guest blogger Søren Hough, a Biochemistry PhD candidate at the University...double-stranded breaks (Tsai et al. 2015). Additionally, current algorithms do not take into account other elements...sequencing (Hendel et al. 2015), a significant difference as CRISPR experiments move closer to the clinic...and Joung 2016. Many thanks to our guest blogger Søren Hough. We additionally thank Monica Sentmanat, Victor...Ajetunmobi for their contributions to this work. Søren Hough is a Biochemistry PhD candidate in the laboratory...https://magazine.scienceforthepeople.org/.     References 1. Fu, Yanfang, et al. "High-frequency off-target..."Quantifying on-and off-target genome editing." Trends in biotechnology 33.2 (2015): 132-140. PubMed PMID...