...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...

...Nishimasu, H., Ran, F. A., Hsu, P. D., Konermann, S., Shehata, S. I., Dohmae, N., Ishitani, R., Zhang, F., &...Levesque, S., Jing, Y., Simpson, D., Zhao, A., Li, H., Yan, W., Goudy, L., Schmidt, R., Solley, S. C., Gilbert...Newby, G. A., Sahin, M., Osborn, M. J., Weissman, J. S., Adamson, B., & Liu, D. R. (2021). Enhanced prime....2021.09.018 Nelson, J. W., Randolph, P. B., Shen, S. P., Everette, K. A., Chen, P. J., Anzalone, A. V....

... 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...

...10.1038/s41589-020-0535-8 Takeda S, Kadowaki S, Haga T, Takaesu H, Mitaku S (2002) Identification of G protein-coupled... 950 different human genes encoding GPCRs (Takeda S et al., 2002) and many downstream complexes involved...

...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...

...effect. Which species or variant of Cas9 ( S. pyogenes , S. aureus etc.) was this gRNA sequence designed...

..., C., Lian, C., Luo, S., Liu, L. D., Zhang, S., Zhang, Y., Wo, Y., Li, N., Wu, S., Gui, T., Wang, B., ....1038/nbt.4172 Kurt, I. C., Zhou, R., Iyer, S., Garcia, S. P., Miller, B. R., Langner, L. M., Grünewald...25217-y Zhao, D., Li, J., Li, S., Xin, X., Hu, M., Price, M. A., Rosser, S. J., Bi, C., & Zhang, X. (2021...Rajakumar, P. D., Prekop, H.-T., Chew, Y. T., Manivannan, S. N., & Chew, W. L. (2021). Programmable C:G to G:C...Replogle, J. M., Xu, A., Sisley, T. A., Weissman, J. S., Adamson, B., & Liu, D. R. (2021). Efficient C•G-...

...P, Sommella A, Iodice C, Cesi G, Simone S, Marrocco E, Rossi S, Giunti M, Palfi A, Farrar GJ, Polishchuk...Toriello E, de Simone S, Colella P, Iodice C, Polishchuk EV, Sommella A, Colecchi L, Rossi S, Simonelli F, Giunti...split vector approaches developed in the early 2000’s which allow researchers to deliver oversized cargo...and hematopoietic stem and progenitor cells (HSPC)s to ~40% expression in the K562 cell line (Bak et al....2009.280 Hirsch ML, Li C, Bellon I, Yin C, Chavala S, Pryadkina M, Richard I, Samulski RJ (2013) Oversized...

...., Alicea, B., Audisio, T. L., Auer, S., Bhardwaj, V., Burgess, S. J., Ferling, I., Gazda, M. A., Hoeppner...Ilangovan, V., Lo, H., Olson, M., Mohamed, S. Y., Sarabipour, S., Varma, A., Walavalkar, K., Wissink, E....Prepare and label a sample that expresses your target(s) at a high level, either naturally or by overexpression... D., Murray, K. D., Engebrecht, J., & Trimmer, J. S. (2019). A toolbox of IgG subclass-switched recombinant...10.17504/protocols.io.bs68nhhw Im, K., Mareninov, S., Diaz, M. F. P., & Yong, W. H. (2019). An Introduction...

...Nogales presented the first detailed picture of Cas9's three-dimensional shape via x-ray crystallography,...Institute and MIT published another report showing Cas9's structural details. "Understanding this structure ...This Protein Could Change Biotech Forever."  David S. Weiss of Emory University summed it up well in that...

...Chang, E. B., Ruscheweyh, H.-J., Sunagawa, S., Mclellan, S. L., Willis, A. D., Comstock, L. E., & Eren.... References and Resources References Demanèche, S., Bertolla, F., Buret, F., Nalin, R., Sailland, A.....67.8.3440-3444.2001 Fogarty, E. C., Schechter, M. S., Lolans, K., Sheahan, M. L., Veseli, I., Moore, R....2024.01.039 Garcillán-Barcia, M. P., Redondo-Salvo, S., & de la Cruz, F. (2023). Plasmid classifications...

...Cui, L., Bikard, D., & Van Teeffelen, S. (2018). Tuning dCas9’s ability to block transcription enables...

...Western Ontario. Adam summarizes Zalatan JG, et al.'s recent paper, "Engineering Complex Synthetic Transcriptional... the oft published metabolic engineering realm of S. cerevisiae and E. coli. The utility of simultaneous...potential of this to control a metabolic network in S. cerevisiae by using three RNA programs to differentially...him on Twitter @AdamChinFatt     References Qi, L. S. et al. Repurposing CRISPR as an RNA-guided platform...

... to light until 2007. Initial experiments exposed S. thermophilus to predatory phages to test if the exogenous...technology? Let us know.  References Ishino Y, Hideo, S, Makino K, Mitsuko A, Nakata A. J Bacteriol. 1987 ...Barrangou R, Brouns SJJ, Charpentier E, Horvath P, Moineu S, Mojica FJM, Wolf YI, Yakunin JvdO, Koonin EV. Nature...2013 Jan; 493, 429–432. Cong L, Ran FA, Cox D, Lin S, Barretto R, Habib N, Hsu PD, Wu X, Jiang W, Marraffini...

...and resources References Dong D, Guo M, Wang S, Zhu Y, Wang S, Xiong Z, Yang J, Xu Z, Huang Z (2017) Structural...Inhibit DNA binding Example:  AcrIIA4 blocks Cas9’s interaction with the PAM site (Dong et al., 2017),...deletion could replicate when challenged with the S. islandicus CRISPR-Cas system. 5. Phage therapy Phage...doi.org/10.1016/j.cell.2017.07.037 Li C, Psatha N, Gil S, Wang H, Papayannopoulou T, Lieber A (2018) HDAd5/...

...Chavez A, Scheiman J, Vora S, Pruitt BW, Tuttle M, P R Iyer E, Lin S, Kiani S, Guzman CD, Wiegand DJ, Ter-Ovanesyan...567 . https://doi.org/10.1038/nmeth.3871 Konermann S, Brigham MD, Trevino AE, Joung J, Abudayyeh OO, Barcena...https://doi.org/10.1016/j.cell.2013.02.022 Sajwan S, Mannervik M (2019) Gene activation by dCas9-CBP and...

... zinc-finger arrays. Maeder ML, Thibodeau-Beganny S, Sander JD, Voytas DF, Joung JK. Nature Protocols ...

...OO, Gootenberg JS, Konermann S, Joung J, Slaymaker IM, Cox DBT, Shmakov S, Makarova KS, Semenova E, Minakhin...-Gonzalez A, Zorn K, Gopez A, Hsu E, Gu W, Miller S, Pan C-Y, Guevara H, Wadford DA, Chen JS, Chiu CY ... Barnes KG, Chak B, Mondini A, Nogueira ML, Isern S, Michael SF, Lorenzana I, Yozwiak NL, MacInnis BL,... . https://doi.org/10.1261/rna.054098.115 Shmakov S, Abudayyeh OO, Makarova KS, Wolf YI, Gootenberg JS...JS, Semenova E, Minakhin L, Joung J, Konermann S, Severinov K, Zhang F, Koonin EV (2015) Discovery and...

...R, Hardman K, Jacobson J, Johnson S, Kaufman B, Lee S, Lee T, Pope S, Riordan G, Whitlow M (1988) Single-chain.../10.1126/science.3140379  Bradbury ARM, Sidhu S, Dübel S, McCafferty J (2011) Beyond natural antibodies...molecules, called Bispecific T-cell engagers (BiTE®s), bind CD3 on T cells and a tumor-specific antigen...