Reproducibility of CRISPR-Cas9 methods for generation of conditional mouse alleles: a multi-center evaluation.
Channabasavaiah Basavaraju GurumurthyAidan R O'BrienRolen M QuadrosJohn AdamsPilar AlcaideShinya AyabeJohnathan BallardSurinder K BatraMarie-Claude BeauchampKathleen A BeckerGuillaume BernasDavid BroughFrancisco Carrillo-SalinasWesley ChanHanying ChenRuby DawsonVictoria DeMambroJinke D'HontKatharine M DibbJames D EudyLin GanJing GaoAmy GonzalesAnyonya R GunturHuiping GuoDonald W HarmsAnne HarringtonKathryn E HentgesNeil HumphreysShiho ImaiHideshi IshiiMizuho IwamaEric JonaschMichelle KarolakBernard KeavneyNay-Chi KhinMasamitsu KonnoYuko KotaniYayoi KunihiroImayavaramban LakshmananCatherine LarochelleCatherine B LawrenceLin LiVolkhard LindnerXian-De LiuGloria Lopez-CastejonAndrew LoudonJenna LoweLoydie A Jerome-MajewskaTaiji MatsusakaHiromi MiuraYoshiki MiyasakaBenjamin MorpurgoKatherine MotylYo-Ichi NabeshimaKoji NakadeToshiaki NakashibaKenichi NakashimaYuichi ObataSanae OgiwaraMariette OuelletLeif OxburghSandra PiltzIlka PinzMoorthy P PonnusamyDavid RayRonald J RedderClifford J RosenNikki RossMark T RuheLarisa RyzhovaAne M SalvadorSabrina Shameen AlamRadislav SedlacekKaran SharmaChad SmithKatrien StaesLora StarrsFumihiro SugiyamaSatoru TakahashiTomohiro TanakaAndrew W TraffordYoshihiro UnoLeen VanhoutteFrederique VanrockeghemBrandon J WillisChristian S WrightYuko YamauchiXin YiKazuto YoshimiXuesong ZhangYu ZhangMasato OhtsukaSatyabrata DasDaniel J GarryTino HochepiedPaul ThomasJan Parker-ThornburgAntony D AdamsonAtsushi YoshikiJean-Francois SchmouthAndrei GolovkoWilliam R ThompsonK C Kent LloydJoshua A WoodMitra CowanTomoji MashimoSeiya MizunoHao ZhuPetr KasparekLucy LiawJoseph M MianoGaetan BurgioPublished in: Genome biology (2019)
We propose that the two-donor method lacks efficiency because it relies on two simultaneous recombination events in cis, an outcome that is dwarfed by pervasive accompanying undesired editing events. The methods that use one-donor DNA are fairly efficient as they rely on only one recombination event, and the probability of correct insertion of the donor cassette without unanticipated mutational events is much higher. Therefore, one-donor methods offer higher efficiencies for the routine generation of cKO animal models.