DATE: November 2, 2017
TIME: 10:00am PDT, 1:00pm EDT

A CRISPR Way of Making Mice

The bulk of genetic variation associated with human disease exists in the noncoding genome, much of which comprises transcribed long noncoding RNAs and non-transcribed regulatory sequences. While headway has been made in elucidating the function of some non-coding sequences, we have much to learn and we know virtually nothing about how sequence variants (such as SNPs) in regulatory sequences contribute to human disease.

CRISPR/Cas-mediated genome editing offers unprecedented ease and efficiency in generating mice carrying small (100s of nucleotides) or large (several kilobases) deletions as well as subtle substitutions of regulatory sequences in their native genomic milieu. Delivering synthetic single guide RNAs (sgRNA) and Cas9 protein (as RNPs) to the 1-cell mouse embryo has resulted in the rapid production of new genetic mouse models that previously would be considered risky or even challenging to engineer.

Optimized approaches to generating, genotyping, and phenotyping ‘CRISPRized’ mice carrying multiple types of genomic sequence edits will be discussed. Data will be presented demonstrating high efficiency editing with synthetic sgRNA leading to nearly 100% success rate of germline transmission, driving rapid generation of mouse models. This new wave of mouse models will accelerate discovery of genomic sequence function, including the role of coding and non-coding sequence variants associated with human disease.

Learning Objectives:

• Explain mouse model generation using CRISPR/Cas9
• Optimized approaches to generating, genotyping, and phenotyping ‘CRISPRized’ mice
• Use of mouse models to study function and disease impact of coding & non-coding sequence variation