A mouse model for adult cardiac-specific gene deletion with CRISPR/Cas9

Kelli J. Carroll, Catherine A. Makarewich, John McAnally, Douglas M. Anderson, Lorena Zentilin, Ning Liu, Mauro Giacca, Rhonda Bassel-Duby, Eric N. Olson

Research output: Contribution to journalArticlepeer-review

131 Scopus citations


Clustered regularly interspaced short palindromic repeats (CRISPR)-associated (Cas)9 genomic editing has revolutionized the generation of mutant animals by simplifying the creation of null alleles in virtually any organism. However, most current approaches with this method require zygote injection, making it difficult to assess the adult, tissue-specific functions of genes that are widely expressed or which cause embryonic lethality when mutated. Here, we describe the generation of cardiac-specific Cas9 transgenic mice, which express high levels of Cas9 in the heart, but display no overt defects. In proof-of-concept experiments, we used Adeno-Associated Virus 9 (AAV9) to deliver single-guide RNA (sgRNA) that targets the Myh6 locus exclusively in cardiomyocytes. Intraperitoneal injection of postnatal cardiac-Cas9 transgenic mice with AAV9 encoding sgRNA against Myh6 resulted in robust editing of the Myh6 locus. These mice displayed severe cardiomyopathy and loss of cardiac function, with elevation of several markers of heart failure, confirming the effectiveness of this method of adult cardiac gene deletion. Mice with cardiac-specific expression of Cas9 provide a tool that will allow rapid and accurate deletion of genes following a single injection of AAV9-sgRNAs, thereby circumventing embryonic lethality. This method will be useful for disease modeling and provides a means of rapidly editing genes of interest in the heart.

Original languageEnglish (US)
Pages (from-to)338-343
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Issue number2
StatePublished - Jan 12 2016


  • CRISPR-associated endonuclease
  • Cardioediting
  • Cardiovascular pathology
  • Gene knockdown
  • Transgenic mouse

ASJC Scopus subject areas

  • General


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