...team replaced S. pyogenes Cas9 in the ABE complex with PAM-variant Cas9 proteins: the S. pyogenes NG-Cas9... TadA and TadA7.10, they designed one set of ABE8’s similarly (ABE8.x-d). In order to assess the effect...created ABE8 constructs using a catalytically “dead” S. pyogenes Cas9 (dC9-ABE8.x-m/d). The dC9-ABE8.x-m/...scope of adenine base editors The most commonly used S. pyogenes Cas9 requires the target site in the genomic...-Cas9 (PAM: NG) to create NG-ABE8.xm/d and the S. aureus Cas9 (PAM: NNGRRT) to create Sa-ABE8.x-m/d. With...NM, Barrera LA, Born DA, Edwards A, Gehrke JM, Lee S-J, Liquori AJ, Murray R, Packer MS, Rinaldi C, Slaymaker...

...C-Y, Gootenberg JS, Konermann S, Trevino AE, Scott DA, Inoue A, Matoba S, Zhang Y, Zhang F (2013) Double.... https://doi.org/10.1038/ncomms5286 Yan M-Y, Li S-S, Ding X-Y, Guo X-P, Jin Q, Sun Y-C (2020) A CRISPR-Assisted...Piganeau M, Renouf B, Renaud J-B, Sallmyr A, Ruis B, Oh S, Tomkinson AE, Hendrickson EA, Giovannangeli C, Jasin...Pryor JM, Strom CN, Mieczkowski P, Burkhalter MD, Oh S, Qaqish BF, Moore DT, Hendrickson EA, Ramsden DA (...

...profiling. Martin S, Söllner C, Charoensawan V, Adryan B, Thisse B, Thisse C, Teichmann S and Wright GJ Molecular...Bartholdson SJ, Bei AK, Theron M, Uchikawa M, Mboup S, Ndir O, Kwiatkowski DP, Duraisingh MT, Rayner JC ...

....1101/2023.03.20.533580 Goodwin, M. S., Croft, C. L., Futch, H. S., Ryu, D., Ceballos-Diaz, C., Liu, X...Truong, V., Rostonics, E., Hernandez, A., Boye, S. L., Boye, S. E., Levites, Y., … Golde, T. E. (2020). Utilizing...producer cells for further processing. What component(s) you need to collect will depend on the serotype, ....1089/hum.2020.120 Lock, M., Alvira, M. R., Chen, S.-J., & Wilson, J. M. (2014). Absolute determination...doi.org/10.1089/hgtb.2013.131 Lock, M., McGorray, S., Auricchio, A., Ayuso, E., Beecham, E. J., Blouin-Tavel...

...protein labeling and signal amplification. Zhou S, Feng S, Brown D, Huang B. PLoS One. 2020 Bo Huang Yellow...editing (Feng Zhang Lab 2015)  Reference Feng, S., Sekine, S., Pessino, V. et al. Improved split fluorescent...oligomerisation. Eastwood T, Baker K, Brooker H, Frank S, Mulvihill DP. FEBS Lett. 2017 Dan Mulvihill mVenus...visualization of endogenous proteins and synapses. Feng S et al. Commun Biol. 2019 Bo Huang Near-Infrared ...protein-protein interactions in living cells. Shao S, Zhang H, Zeng Y, Li Y, Sun C, Sun Y. Commun Biol....

... B.-Y., Viswanathan, S., Gaj, T., Lavzin, M., Ritola, K.D., Lindo, S., Michael, S., Kuleshova, E., Ojala...complications. References Aschauer, D.F., Kreuz, S., and Rumpel, S. (2013). Analysis of transduction efficiency...Okada, T., Kawasaki, K., Ishibashi, H., Yuasa, S., Takeda, S., Hasegawa, I., and Nakahara, K. (2011). Local...PubMed Central PMCID: PMC2293646. Jin, J., and Maren, S. (2015). Prefrontal-Hippocampal Interactions in Memory...Mise, N., Obata, Y., Wickersham, I.R., and Tonegawa S. (2014). Cell type-specific genetic and optogenetic...22196330. PubMed Central PMCID: PMC3275419. Maday, S., Twelvetrees, A.E., Moughamian, A.J., and Holzbaur...Harris, J.A., Ng, L., Winslow, B., Cain, N., Mihalas, S., Wang, Q., Lau, C., Kuan, L., Henry, A.M., Mortrud...

..., Valois, E., Solley, S. C., Braig, F., Lach, R. S., Baxter, N. J., Kosik, K. S., Arias, C., Acosta-Alvear...coronavirus using CRISPR-based SHERLOCK (Specific ​H​igh ​s​ensitivity ​E​nzymatic ​R​eporter un​LOCK​ing) technique... amplification kit. In this case, regions of gene S and Orf1ab gene are amplified. RPA reactions also ...A. H., Sinha, D., Sharma, N., Kumar, M., … Maiti, S. (2020). Rapid, field-deployable nucleobase detection...-Gonzalez A, Zorn K, Gopez A, Hsu E, Gu W, Miller S, Pan C-Y, Guevara H, Wadford DA, Chen JS, Chiu CY ...2020.04.20.052159 Zhang, F., Abudayyeh, O. O. & Gootenberg, J. S. A protocol for detection of COVID-19 using CRISPR...

...wondered if it could overcome some of Cas9’s shortcomings. Cpf1’s preferred PAM is 5’-TTN, differing from...nucleotide 5’ overhang 18-23 bases away from the PAM. Cpf1’s staggered cleavage pattern opens up the possibility...deletions at the 3’ end of the target sequence. Cpf1’s strength may lie in its low off-target editing rates...with SpCas9. Kleinstiver et al. suggest that AsCpf1’s off-target rate is similar to that of high fidelity...Brouns SJ, Charpentier E, Haft DH, Horvath P, Moineau S, Mojica FJ, Terns RM, Terns MP, White MF, Yakunin ...

...Cho H-Y, Song DW, Lee KJ, Jung MH, Kim S, Kim JH, Kim JH, Kim J-S (2017) In vivo genome editing with a ...doi.org/10.1038/nmeth.3993  Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X, Jiang W, Marraffini...Boyaval P, Fremaux C, Horvath P, Magadán AH, Moineau S (2010) The CRISPR/Cas bacterial immune system cleaves...019-0908-x  Nishimasu H, Ran FA, Hsu PD, Konermann S, Shehata SI, Dohmae N, Ishitani R, Zhang F, Nureki...doi.org/10.1038/nature14299  Senís E, Fatouros C, Große S, Wiedtke E, Niopek D, Mueller A-K, Börner K, Grimm...doi.org/10.1002/biot.201400046  Strecker J, Jones S, Koopal B, Schmid-Burgk J, Zetsche B, Gao L, Makarova...

...Chavez A, Scheiman J, Vora S, Pruitt BW, Tuttle M, P R Iyer E, Lin S, Kiani S, Guzman CD, Wiegand DJ, Ter-Ovanesyan...deliver the appropriate isoform(s) of a given gene, as, many times, the isoform(s) common to a particular cell...Santos R, Rao Y, Sassi F, Pinnelli M, Ansari R, Harper S, Jackson DA, McRae R, Pooley R, Wilkinson P, van der...https://doi.org/10.1016/j.cell.2014.09.029 Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena...

...Scheff, N. N., Snyder, L. M., Ross, S. E., Davis, B. M., & Gold, M. S. (2016). Gi-DREADD Expression in Peripheral... JC, Snowball A, Knauss S, von Schimmelmann M, During MJ, Lignani G, Schorge S, Young D, Kullmann DM, ...Goutaudier, R., Coizet, V., Carcenac, C., & Carnicella, S. (2019). DREADDs: The Power of the Lock, the Weakness..., Hu X, Galvan A, Basu J, Michaelides M, Sternson S (2019) Ultrapotent chemogenetics for research and ...

...Ayoubi, R., Ryan, J., Biddle, M. S., Alshafie, W., Fotouhi, M., Bolivar, S. G., Ruiz Moleon, V., Eckmann,...https://doi.org/10.1038/nmeth.3995 Virk, H. S., Biddle, M. S., Smallwood, D. T., Weston, C. A., Castells.../doi.org/10.1111/bph.15467 Virk, H. S., Rekas, M. Z., Biddle, M. S., Wright, A. K. A., Sousa, J., Weston...RP91645. https://doi.org/10.7554/eLife.91645 Biddle, M. S., Alende, C., Fotouhi, M., Jones, C., Ayoubi, R., ... M., Healy, L. M., Robertson, J., & McPherson, P. S. (2019). Implementation of an antibody characterization...10.7554/eLife.48363 Uhlen, M., Bandrowski, A., Carr, S., Edwards, A., Ellenberg, J., Lundberg, E., Rimm, ..., Bowman, V. W., McCarthy, J., Amrani, Y., Duffy, S. M., Bradding, P., & Roach, K. M. (2021). TGFβ1 induces...

...another important step forward by dissecting that S. pyogenes protein into three parts. Their efforts ...protein, which they call SpyLigase (Spy comes from the “S” in Streptococcus and the “py” in pyogenes), capable... cell capture. Fierer et al (Proc Natl Acad Sci U S A. 2014 Mar 17. PubMed) Peptide tag forming a rapid...bacterial adhesin. Zakeri et al (Proc Natl Acad Sci U S A. 2012 Mar 20;109(12):E690-7. Epub 2012 Feb 24. PubMed...

...Cloning Enzyme(s) Validated In Resistance Co-expressed Cas9 Depositing lab Cas9 species = S. pyogenes (PAM...PAM = NGG) pRPR1_gRNA_ handle_RPR1t pRPR1 HindIII S. cerevisiae LEU2 none, need Cas9 plasmid Lu Do you...

... B. S., Purohit, A. M., Grippo, R. M., Gaykema, R. P., Spano, A. J., Beenhakker, M. P., Kucenas, S., Patel...References and resources References Duret, G., Polali, S., Anderson, E. D., Bell, A. M., Tzouanas, C. N., Avants...andrewgyork.github.io/gfp_magnetofluorescence. Karki, N., Vergish, S., & Zoltowski, B. D. (2021). Cryptochromes: Photochemical...Déjean, V., Dodson, C. A., Wallace, M. I., Mackenzie, S. R., Timmel, C. R., & Hore, P. J. (2016). Chemical...doi.org/10.1021/nl202950h.  Matsuoka, R., Kimura, S., Miura, T., Ikoma, T., & Kusamoto, T. (2023) Single-Molecule...

...K., Kim, Y., Lee, S., Min, S., Bae, J. Y., Choi, J. W., Park, J., Jung, D., Yoon, S., & Kim, H. H. (2019... Kim, H., Lalanne, J.-B., Nathans, J. F., Lee, D. S., & Shendure, J. (2024). Chromatin context-dependent..., Schep, R., Damodharan, T., Balázs, Z., Janjuha, S., Ioannidi, E. I., Böck, D., Steensel, B. van, Krauthammer... Weide, R. H., Morris, B., van Schaik, T., Manzo, S. G., Peric-Hupkes, D., van den Berg, J., Beijersbergen...

...spacer(s) into the CRISPR array Clone desired DNA cargo into the mini-transposon Transform plasmid(s) into...Dong F, Yang S, Yang J, Zhang Z, Qian Y, Chen J, Zhang J, Liu Y, Tao R, Jiang Y, Yang J, Yang S (2020) Multicopy...s41586-019-1323-z Peters JE, Makarova KS, Shmakov S, Koonin EV (2017) Recruitment of CRISPR-Cas systems...doi.org/10.1073/pnas.1709035114 Rubin BE, Diamond S, Cress BF, Crits-Christoph A,  He C , Xu M, Zhou Z...

...pPLPCB(s). pJT118 31395 PcpcG2 Green Light (532 nm) Christopher Voigt When combined with pPLPCB(S), encodes...Anhydrotetracycline inducible expression of wild-type Cas9 from S. pyogenes for inducing double stranded breaks. For...and interactions! Plasmid ID Promoter Epitope Tag(s) ID Purpose pDEST-HisMBP 11085 Tac (lactose/IPTG inducible.../protein of interest. Plasmid ID Promoter Inducer(s) PI Purpose Murray Lab pBEST plasmids Various Plasmids...inducible expression of catalytically inactive Cas9 ( S. pyogenes ) which can be combined with expression ...Christopher Voigt When combined with pCph8 and pPLPCB(S), makes lacZ expression inducible by red light. pCph8...

...Oldewurtel E, Cui L, Bikard D, van Teeffelen S. Tuning dCas9’s ability to block transcription enables robust...

...Rohner, V., Kiss, C., Curreli, S., Dieter, A., Wilhelm, M., Harada, M., Duss, S. N., Dernic, J., Bhat, M. ... Z., Wu, H., Chen, W., Zhang, P., Zou, J., Hires, S. A., Zhu, J. J., Cui, G., Lin, D., Du, J., & Li, Y... Schütze, K., Moriwaki, Y., Heo, L., Ovchinnikov, S., & Steinegger, M. (2022). ColabFold: Making protein...