...responsible for that differentiation.   Fig. 2: Single cell RNA-seq clustering of cells transduced...RNA-seq was performed to map TFs to cell fates. (Adapted from Joung et al.)    This atlas linked TF ... color coded nominated reference cell types.  (Adapted from Joung et al.)    Putting MORF to work ...

...retroviral library was used to transduce T helper type 2 (Th2) cells from mice that constitutively express ...Nature Communications 2019. Pubmed PMID: 31366908 Adapting sigma54-dependent transcription for CRISPR activation...must be σ54-dependent and the system may not be adaptable to distantly related species from E. coli. This...

...Enhanced CRISPR-based DNA demethylation Figure 2: The components of Casilio-ME1. Image from Taghbalout...ctrA gene expression using various dCas9. Image adapted from Guzzo et al., 2020. Caulobacter crescentus...

...Genome Editing and The CRISPR Software Matchmaker Day 2: Homology directed repair: CRISPR 101: Homology Directed...? I certainly didn’t, but now we can with Cas13b/ADAR constructs from the REPAIR (RNA Editing for Programmable...

...156–159. https://doi.org/10.1016/0003-2697(87)90021-2 Chomczynski P, Sacchi N (2006) The single-step method...groups are susceptible to hydrolysis and heat degradation. Furthermore, RNases, or enzymes that degrade...samples cold and work quickly to mitigate RNA degradation. Make sure to use DEPC-treated or RNAse free ...store them in a -80°C freezer to prevent RNA degradation. They will degrade in a -20°C freezer.   RNA...

...unc- 119 + selection." MicroPublication 1 (2018): 2–3. Wang, Han, et al. "cGAL, a temperature-robust GAL4...for the Auxin Inducible Degradation System (AID) The Dernberg lab in 2015 adapted the AID system discovered...and new tools for conditional expression and degradation. These workshops not only highlighted new tools...  Zhang, Liangyu, et al. "The auxin-inducible degradation (AID) system enables versatile conditional protein...

...systems utilize with type I-F Cascades. Figure 2: Cascade complex from CRISPR type I-E. Created with...binding and subsequent DNA degradation appears to be conserved. This DNA degradation portion can be bypassed.... These alterations are achieved through DNA degradation, thanks to the unique capabilities of Cas3 and...of DNA, Cascade caught up to it, and then DNA degradation continued. However, Cas3 cannot bind DNA on its...Hopefully more people will want to utilize Cas3’s degradative power in the future! References and Resources.... (2018). Structure basis for RNA-guided DNA degradation by Cascade and Cas3. Science, 361(6397). https...., Sonpho, E., Xu, H., Sasakawa, N., Naito, Y., Nakada, S., Yamamoto, T., Sano, S., Hotta, A., Takeda,...

...backbone, which expresses the gRNA from a Drosophila U6:2 promoter and Cas9 from the actin 5C promoter. Addgene...backbones for lentivirus LV 1-5 and retrovirus RV 2-5 were constructed. The authors then used GMAP assembly...engineering (MAGE) (1). MAGE uses recombineering (2) to simultaneously incorporate multiple single-strand...number of tandemly expressed gRNAs respectively) and 2) the size of the deletion. Smaller deletions generally...catalytic domain of RNA editing enzyme, ADAR. This RBP-ADARcd fusion protein irreversibly edits adenine...unsuspected genes with known roles in ER-associated degradation (ERAD) that positively regulate the ability of...2015. PubMed PMID: 26264872 Rapid, conditional degradation of essential proteins using Cas9-based AID tagging...

...project my team & I have poured our hearts into for 2 yrs) has 200K+ figures by 95K+ scientists 👨🏽‍🔬👩‍...workspaces at a time and workspaces are only downloadable in a small file size with a Biorender watermark...

...development and therapy. Nat Rev Drug Discov (2017) 16(2):89-100. DOI: 10.1038/nrd.2016.238 Additional resources...precise disease genetics. Since CRISPR has been adapted for use in primary cells, many tissue types, and...

...1) or added using PCR-based amplification (Option 2). Advantages of Golden Gate cloning Golden Gate cloning...few steps. More recently, CRISPR technology has adapted Golden Gate cloning for inserting the appropriate...single gRNA-expressing plasmid, but it can also be adapted to express multiple gRNAs. Addgene has two Golden...

...inhibit editing (Anzalone et al., 2019). Figure 2: Starting lengths for optimizing your pegRNA primer... motifs at their 3′ end to protect them from degradation. When designing epegRNAs with large structured...different strategies for protecting pegRNAs from degradation. Created with BioRender.com. That’s all ...A., Sahin, M., Osborn, M. J., Weissman, J. S., Adamson, B., & Liu, D. R. (2021). Enhanced prime editing...., Bauer, D. E., Marson, A., Parsons, L. R., & Adamson, B. (2024). Improving prime editing with an endogenous...

... 2008 Dec; 18(6): 617–623. Pubmed PMID 19349161.  2. Deshpande A, Bergami M, Ghanem A, Conzelmann KK, ...only infect Cre-positive, TVA-expressing cells (Osakada et al., 2013). Ultimately, this allows RABV to ...Pubmed PMID 23487772. 3. Etessami R, Conzelmann KK, Fadai-Ghotbi B, Natelson B, Tsiang H, Ceccaldi PE. Spread...Neuroanat. 2015 Jul 1;9:80. Pubmed PMID 26190977. 6. Osakada F, Callaway EM. Design and generation of recombinant...

... the Cas machinery where to cut.      Fig. 2: Heatmap indicating loci where SNPs reside on sgRNA... Lia Petronio, Katherine Huang, Alham Saadat, Thomas Green, Adam Brown, John G Doench, David E Root, James...worked with John Doench, David Root, Tom Green, and Adam Brown from Broad's Genetic Perturbation Platform...

...containing complementary polynucleotide chains. J Virol 9(2): 394-396. Lecomte E, Tournaire B, Cogné B, Dupont... After launching our viral service in 2016, we adapted this platform to accommodate adeno-associated viral...

...extraction protocol for more details.   Figure 2: An uncut plasmid is expected to form 3 bands: nicked...plasmids and testing your prep can save you the headache of troubleshooting failed experiments. Pick the...

...MagFRET: The First Genetically Encoded Fluorescent Mg(2+) Sensor. PLoS One. 2013 Dec 2;8(12):e82009. doi: ...to measure them. Merkx says, while in principle adapting a FRET sensor for monitoring calcium versus zinc...

...C-terminus of human codon-optimized  cryptochrome 2 (CRY2) from Arabidopsis thaliana. When exposed to ...containing a synthetic operon that enables the degradation of caffeine (1,3,7-trimethylxanthine) and its...protein-protein interactions. We’re excited to see how you adapt similar FLINC biosensors to work in your lab. Find...

...interferon gamma (IFNγ) - and eliminate cancer cells. 2. Equilibrium - In a process analogous to the development... 35. Academic Press, 2015. PubMed PMID: 25818339. 2. Curiel, Tyler J., et al. "Specific recruitment of... radiation therapy (owing to contributions from Madame Curie), chemotherapy, and surgical removal of the...

...two options to begin your plasmid deposit. Figure 2: Choose to submit plasmid(s) online or by using a ...format. Figure 6: Add sequence, maps, and files. Step 2: Gene and Insert *Note: This page will not appear ... entered using our online form or by using a downloadable spreadsheet. Either choice can be used for deposits...