... of new a tool for CRISPR scientists––CrispyCrunch! CrispyCrunch is a web app that helps scientists design...works, and how to use it. Background on CrispyCrunch CrispyCrunch was initially built for the CZ Biohub ...internally. Guiding principles behind CrispyCrunch While CrispyCrunch may evolve over time, you can depend...mutation in CrispyCrunch: If the guide PAM resides outside a coding region in a UTR, CrispyCrunch flips the...chosen for a silent mutation.) How to use CrispyCrunch CrispyCrunch take you through the whole process from...information on using CrispyCrunch, be sure to check out the How To Guide. Conclusion CrispyCrunch is now publicly-available...alleviate this bottleneck, we began building CrispyCrunch in July 2018 and in November, we began using...

... efforts yielded a protein, which they call SpyLigase (Spy comes from the “S” in Streptococcus and the... to develop SpyLigase?  Howarth: It came from our work in developing the SpyTag/SpyCatcher system, to ... particular. They recently introduced the SpyTag/SpyCatcher system, based on a protein isolated from Streptococcus...genetically encoded way. SpyTag is nice and small, just 13 amino acids, but SpyCatcher is a protein. You can...capable of locking two peptide tags together. SpyLigase overcomes limitations in the use of peptide tags...already demonstrated in their PNAS paper that SpyLigase can be used to link affibodies or antibodies against...antigen. I asked Howarth to tell us more about SpyLigase, its development, and its potential uses. Developing...

...type II CRISPR systems, which include S. pyogenes (Spy) Cas9, been identified. In Rauch, et al. 2016 the...proteins, AcrIIA2 and AcrIIA4, can work to inhibit Spy Cas9 function in both bacteria and mammalian cells...

... pJDS Series   Superglue proteins with SpyTag/SpyCatcher and SnoopTag/SnoopCatcher The development ...combination with their previously described SpyTag/SpyCatcher pair, multiple protein fusions can be synthesized... the use of the SnoopTag/SnoopCatcher and SpyTag/SpyCatcher pairs. In order to improve the ease and convenience...MBPx-SpyCatcher construct. This construct is used to produce a maltose binding protein/SpyCatcher fusion...solid-phase synthesis of larger fusions through SpyTag/SpyCatcher and SnoopTag/SnoopCatcher linkages. Once all...stable and directional protein fusion, with a SpyCatcher-SnoopCatcher linker remaining between each protein...

...peptide-peptide ligation using SpyLigase Building off their SpyTag/SpyCatcher system for protein-peptide...between 2 peptide tags, SpyTag and KTag. The group demonstrated the use of the SpyLigase peptide-peptide interaction...peptide-peptide locking. The new technology is known as SpyLigase and is a protein domain that promotes the formation...20;109(12):E690-7. For further reading about SpyLigase technology, read the Addgene interview with Mark...

...beads through an assembly of biotinylated SpyTag-SpyCatcher linkers and spacer proteins ending with a ... single alpha helix) and target capture module (SpyTag-GFP nanobody) assembled on beads. C) Micrograph...miniCRISPR” version, are much smaller proteins than SpyCas9, allowing them to be packaged in AAV vectors, potentially...

...Thomas Huber 182287 pcDNA3.1(+)_U6 tRNAPyl_CMV NESPylRS(AF) Y306A/Y384F (AF) pyrrolysine (Pyl) tRNA synthetase...TAA Wenshe Liu 182653 pcDNA3.1(+)_U6 tRNAPyl_CMV NESPylRS(AF)_IRES_eRF1(E55D)-HA Y306A/Y384F (AF) pyrrolysine...azidonorleucine (Anl) Mammalian Elizabeth Kirby 223508 pCMV_NESPylRS(AF)_hU6tRNAPyl PylRS (Y306A, Y384F) M. mazei...mazei Mammalian TAG Edward Lemke 223509 pCMV_NESPylRS(WT)_hU6tRNAPyl PylRS M. mazei Mammalian TAG Edward ...

...and trans activity of each entry. For example, SpyCas9a (one of the most frequently used Cas enzymes) ...CasPEDIA Cas ID display information for a given Cas (SpyCas9a) from CasPEDIA.   Helpful resources and information...List of resources and properties for a Cas entry (SpyCas9a) from CasPEDIA.   How can I use it? CasPEDIA...

...lentiviral plasmids, catalytically inactive Cas9 (dSpyCas9) fused to APEX2 is expressed from a Tet/Shield1...are enriched in the 20 nm radius surrounding the dSpyCas9-APEX2 fusion.  This Sontheimer lab used this technology...

... selected sgRNAs, genes, or pathways. CrispyCrunch CrispyCrunch is used for high-throughput design and...

...al., 2016). While Streptococcus pyogenes Cas9 (SpyCas9) is the most used and well studied CRISPR system...Xu Z, Huang Z (2017) Structural basis of CRISPR–SpyCas9 inhibition by an anti-CRISPR protein. Nature 546...

... an overlapping PAM. In both cases, GeoCas9 and SpyCas had similar levels of editing. See figure 4 in ...

...epitope tags (HA, Myc, Strep tag II, AviTag, HaloTag, SpyTag). The Foerstemann lab developed a polymerase chain...

... regulated by guanosine [corrected] 3', 5'-bispyrophosphate: a model for programmed bacterial cell death...

...lab CRISPR Challenges: Standardization and HDR CrispyCrunch Optimizing Donor DNA for Enhanced CRISPR Genome...