Response to correspondence on "Reproducibility of CRISPR-Cas9 methods for generation of conditional mouse alleles: a multi-center evaluation"

• Published in: Genome Biology 2021-04, Vol.22 (1), p.99-99, Article 99
• Main Authors: Gurumurthy, Channabasavaiah B, O'Brien, Aidan R, Quadros, Rolen M, Adams, Jr, John, Alcaide, Pilar, Ayabe, Shinya, Ballard, Johnathan, Batra, Surinder K, Beauchamp, Marie-Claude, Becker, Kathleen A, Bernas, Guillaume, Brough, David, Carrillo-Salinas, Francisco, Chan, Wesley, Chen, Hanying, Dawson, Ruby, DeMambro, Victoria, D'Hont, Jinke, Dibb, Katharine, Eudy, James D, Gan, Lin, Gao, Jing, Gonzales, Amy, Guntur, Anyonya, Guo, Huiping, Harms, Donald W, Harrington, Anne, Hentges, Kathryn E, Humphreys, Neil, Imai, Shiho, Ishii, Hideshi, Iwama, Mizuho, Jonasch, Eric, Karolak, Michelle, Keavney, Bernard, Khin, Nay-Chi, Konno, Masamitsu, Kotani, Yuko, Kunihiro, Yayoi, Lakshmanan, Imayavaramban, Larochelle, Catherine, Lawrence, Catherine B, Li, Lin, Lindner, Volkhard, Liu, Xian-De, Lopez-Castejon, Gloria, Loudon, Andrew, Lowe, Jenna, Jerome-Majeweska, Loydie, Matsusaka, Taiji, Miura, Hiromi, Miyasaka, Yoshiki, Morpurgo, Benjamin, Motyl, Katherine, Nabeshima, Yo-Ichi, Nakade, Koji, Nakashiba, Toshiaki, Nakashima, Kenichi, Obata, Yuichi, Ogiwara, Sanae, Ouellet, Mariette, Oxburgh, Leif, Piltz, Sandra, Pinz, Ilka, Ponnusamy, Moorthy P, Ray, David, Redder, Ronald J, Rosen, Clifford J, Ross, Nikki, Ruhe, Mark T, Ryzhova, Larisa, Salvador, Ane M, Alam, Sabrina Shameen, Sedlacek, Radislav, Sharma, Karan, Smith, Chad, Staes, Katrien, Starrs, Lora, Sugiyama, Fumihiro, Takahashi, Satoru, Tanaka, Tomohiro, Trafford, Andrew, Uno, Yoshihiro, Vanhoutte, Leen, Vanrockeghem, Frederique, Willis, Brandon J, Wright, Christian S, Yamauchi, Yuko, Yi, Xin, Yoshimi, Kazuto, Zhang, Xuesong, Zhang, Yu, Ohtsuka, Masato, Das, Satyabrata, Garry, Daniel J, Hochepied, Tino, Thomas, Paul, Parker-Thornburg, Jan, Adamson, Antony D, Yoshiki, Atsushi
• Format: Article
• Language: English
• Subjects: Alleles; Animals; Correspondence; CRISPR; CRISPR-Associated Protein 9 - metabolism; CRISPR-Cas Systems; Efficiency; Gene Editing; Gene loci; Genetic engineering; Immunology; Laboratories; Medical research; Medicine; Methods; Mice; Molecular biology; Nervous system; Neurosciences; Reproducibility; Reproducibility of Results; Research centers; Science; Transgenic mice
• ISSN: 1474-760X; 1474-7596; 1474-760X
• DOI: 10.1186/s13059-021-02320-3

The two Jaenisch laboratory studies published in Cell in 2013 were ground-breaking, demonstrating for the first time proof of principle CRISPR mediated gene editing in the mouse zygote to generate knockout and conditional alleles, and caused much excitement in the transgenic mouse community. [...]our study is not the first time concerns have been raised as to the efficiency of the 2-guides 2-oligo method, with anecdotal reports from others in the transgenic community (Science; 2016. doi:https://doi.org/10.1126/science.aal0334 [doi.org]), which stated that “What was disappointing is none of us could reproduce at the efficiencies reported by Jaenisch. Because efficiencies at different genomic loci often vary highly (which the Yang et al. authors state in their paragraph below point #3), it would be ideal to gather such side-by-side data for at least 6 to 10 loci or more to ensure reproducibility. [...]our observations call into question the robustness of the approach and its suitability for widespread use. Division of Neuroscience and Experimental Psychology, School of Biological Sciences, Faculty of Biology, Medicine and Health, Manchester Academic Health Science Centre, University of Manchester, AV Hill Building, Oxford Road, Manchester, M13 9PT, UK David Brough & Catherine B. Lawrence 13.