Gene Editing with CRISPR/Cas9 RNA-Directed Nuclease

Thomas C Doetschman, Teodora Georgieva

Research output: Contribution to journalReview article

20 Citations (Scopus)

Abstract

Genetic engineering of model organisms and cultured cells has for decades provided important insights into the mechanisms underlying cardiovascular development and disease. In the past few years the development of several nuclease systems has broadened the range of model/cell systems that can be engineered. Of these, the CRISPR (clustered regularly interspersed short palindromic repeats)/Cas9 (CRISPR-associated protein 9) system has become the favorite for its ease of application. Here we will review this RNA-guided nuclease system for gene editing with respect to its usefulness for cardiovascular studies and with an eye toward potential therapy. Studies on its off-target activity, along with approaches to minimize this activity will be given. The advantages of gene editing versus gene targeting in embryonic stem cells, including the breadth of species and cell types to which it is applicable, will be discussed. We will also cover its use in iPSC for research and possible therapeutic purposes; and we will review its use in muscular dystrophy studies where considerable progress has been made toward dystrophin correction in mice. The CRISPR/Ca9s system is also being used for high-throughput screening of genes, gene regulatory regions, and long noncoding RNAs. In addition, the CRISPR system is being used for nongene-editing purposes such as activation and inhibition of gene expression, as well as for fluorescence tagging of chromosomal regions and individual mRNAs to track their cellular location. Finally, an approach to circumvent the inability of post-mitotic cells to support homologous recombination-based gene editing will be presented. In conclusion, applications of the CRISPR/Cas system are expanding at a breath-taking pace and are revolutionizing approaches to gain a better understanding of human diseases.

Original languageEnglish (US)
Pages (from-to)876-894
Number of pages19
JournalCirculation Research
Volume120
Issue number5
DOIs
StatePublished - Mar 3 2017

Fingerprint

Ribonucleases
Long Noncoding RNA
Therapeutic Human Experimentation
Dystrophin
Genetic Engineering
Gene Targeting
Muscular Dystrophies
Nucleic Acid Regulatory Sequences
Homologous Recombination
Genetic Models
Embryonic Stem Cells
Genes
Cultured Cells
Cardiovascular Diseases
Fluorescence
Gene Expression
Messenger RNA
Gene Editing
Proteins
Therapeutics

Keywords

  • cardiovascular diseases
  • clustered regularly interspaced short palindromic repeats
  • gene editing
  • genetic engineering
  • muscular dystrophies

ASJC Scopus subject areas

  • Physiology
  • Cardiology and Cardiovascular Medicine

Cite this

Gene Editing with CRISPR/Cas9 RNA-Directed Nuclease. / Doetschman, Thomas C; Georgieva, Teodora.

In: Circulation Research, Vol. 120, No. 5, 03.03.2017, p. 876-894.

Research output: Contribution to journalReview article

Doetschman, Thomas C ; Georgieva, Teodora. / Gene Editing with CRISPR/Cas9 RNA-Directed Nuclease. In: Circulation Research. 2017 ; Vol. 120, No. 5. pp. 876-894.
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