Phenotyping first-generation genome editing mutants: a new standard?

The unprecedented efficiency of the CRISPR/Cas9 system in genome engineering has opened the prospect of employing mutant founders for phenotyping cohorts, thus accelerating research projects by circumventing the requirement to generate cohorts using conventional two- or three-generation crosses. However, these first-generation mutants are often genetic mosaics, with a complex and difficult to define genetic make-up. Here, we discuss the potential benefits, challenges and scientific validity of such models.

[1]  R. Palmiter,et al.  Germline and somatic mosaicism in transgenic mice. , 1986, Developmental biology.

[2]  S. Honma,et al.  Disruption of MeCP2 attenuates circadian rhythm in CRISPR/Cas9‐based Rett syndrome model mouse , 2015, Genes to cells : devoted to molecular & cellular mechanisms.

[3]  J. Parant,et al.  High-Throughput Genome Editing and Phenotyping Facilitated by High Resolution Melting Curve Analysis , 2014, PloS one.

[4]  Steve D. M. Brown,et al.  High-throughput discovery of novel developmental phenotypes , 2017 .

[5]  J. Rossant,et al.  Chimeras and mosaics in mouse mutant analysis. , 1998, Trends in genetics : TIG.

[6]  Rudolf Jaenisch,et al.  One-Step Generation of Mice Carrying Mutations in Multiple Genes by CRISPR/Cas-Mediated Genome Engineering , 2013, Cell.

[7]  R. Jaenisch,et al.  One-Step Generation of Mice Carrying Reporter and Conditional Alleles by CRISPR/Cas-Mediated Genome Engineering , 2013, Cell.

[8]  R. Behringer,et al.  New mouse models of cancer: Single-cell knockouts , 2007, Proceedings of the National Academy of Sciences.

[9]  G. Mardon,et al.  CRISPR-engineered mosaicism rapidly reveals that loss of Kcnj13 function in mice mimics human disease phenotypes , 2015, Scientific Reports.

[10]  Yi Zhou,et al.  A CRISPR/Cas9 vector system for tissue-specific gene disruption in zebrafish. , 2015, Developmental cell.

[11]  B. A.,et al.  Disease model discovery from 3,328 gene knockouts by The International Mouse Phenotyping Consortium , 2018, Yearbook of Paediatric Endocrinology.

[12]  I. Cuthill,et al.  Animal Research: Reporting In Vivo Experiments: The ARRIVE Guidelines , 2010, British journal of pharmacology.

[13]  S. Wells,et al.  Analysing the outcome of CRISPR-aided genome editing in embryos: Screening, genotyping and quality control. , 2017, Methods.

[14]  Steve D. M. Brown,et al.  Disease Model Discovery from 3,328 Gene Knockouts by The International Mouse Phenotyping Consortium , 2017, Nature Genetics.

[15]  Steve D. M. Brown,et al.  Correction of the auditory phenotype in C57BL/6N mice via CRISPR/Cas9-mediated homology directed repair , 2016, Genome Medicine.

[16]  N. Perrimon.  Creating mosaics in Drosophila. , 1998, The International journal of developmental biology.

[17]  Joseph S. Takahashi,et al.  Chimera Analysis of the Clock Mutation in Mice Shows that Complex Cellular Integration Determines Circadian Behavior , 2001, Cell.

[18]  Steve D. M. Brown,et al.  Prevalence of sexual dimorphism in mammalian phenotypic traits , 2017, Nature Communications.

[19]  Simon Watkins,et al.  The complex genetics of hypoplastic left heart syndrome , 2017, Nature Genetics.

[20]  A. Bradley,et al.  Induced mitotic recombination of p53 in vivo , 2007, Proceedings of the National Academy of Sciences.

[21]  Jean-Paul Concordet,et al.  Improved Genome Editing Efficiency and Flexibility Using Modified Oligonucleotides with TALEN and CRISPR-Cas9 Nucleases. , 2016, Cell reports.

[22]  Colin Smith,et al.  Biallelic mutations in the gene encoding eEF1A2 cause seizures and sudden death in F0 mice , 2017, Scientific Reports.

[23]  G. Ertzeid,et al.  The impact of ovarian stimulation on implantation and fetal development in mice. , 2001, Human reproduction.

[24]  Ewelina Bolcun-Filas,et al.  A Mouse Geneticist’s Practical Guide to CRISPR Applications , 2014, Genetics.

[25]  A. Strongin,et al.  Identification of Annexin A4 as a hepatopancreas factor involved in liver cell survival. , 2014, Developmental biology.

[26]  Laurie D. Smith,et al.  MMP21 is mutated in human heterotaxy and is required for normal left-right asymmetry in vertebrates , 2015, Nature Genetics.

[27]  Min Zhang,et al.  Somatic mosaicism and allele complexity induced by CRISPR/Cas9 RNA injections in mouse zygotes. , 2014, Developmental biology.

[28]  Genshiro A. Sunagawa,et al.  Mammalian Reverse Genetics without Crossing Reveals Nr3a as a Short-Sleeper Gene. , 2016, Cell reports.

[29]  Martin J. Aryee,et al.  GUIDE-Seq enables genome-wide profiling of off-target cleavage by CRISPR-Cas nucleases , 2014, Nature Biotechnology.