Enhancing stem cell survival in an ischemic heart by CRISPR-dCas9-based gene regulation

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

Ischemic heart disease has remained the number one killer around the world for over the past 20. years. While stem cell therapy has become a promising new frontier to repair the damaged heart, limited stem cell survivability post-transplantation has precluded widespread use of this therapy. Strategies to genetically modify stem cells to activate pro-survival and anti-apoptotic and anti-inflammatory pathways, such as Akt and heme oxygenase-1, have been shown to improve the lifespan of transplanted stem cells within the ischemic myocardium, but constitutive overexpression of these pathways at high levels has been shown to have side effects. Therefore, more specific and controlled gene activation would be necessary. Current techniques used for gene regulation include zinc finger and TALE proteins, but there are still disadvantages to each of these methods, such as ease and cost of use. Also, those methods use synthesized promoters to express synthesized cDNA, which lack regulatory elements, including introns and 3' untranslated regions for microRNA mediated post-transcriptional regulation. A new novel technique, the CRISPR/dCas9 system, was recently developed as a simple and efficient method for endogenous gene regulation. With its use of single guide chimeric RNA's (sgRNA's), this system has been shown to provide a high level of specificity and efficiency. When targeting different loci, past studies have found that the CRISPR/dCas9 system can activate gene expression at varying levels. In addition, this system makes use of the genome's endogenous regulatory elements, such as the aforementioned introns and 3' UTR's, which can help provide a safer method of gene activation. If targeted to a gene promoting cellular survival or decreasing cell death, it could potentially improve stem cell longevity in a more efficient and controllable manner. As a result, our hypothesis is to use the CRISPR/dCas9 system to activate expression of an anti-inflammatory and anti-apoptotic gene, such as heme oxygenase-1 (HO-1), to an optimal level to increase transplanted stem cell survival while also mitigating its cytotoxic effects due to lack of internal regulation, thus prolonging its effects within the ischemic myocardium leading to greater therapeutic benefit.

Original languageEnglish (US)
Pages (from-to)702-705
Number of pages4
JournalMedical Hypotheses
Volume83
Issue number6
DOIs
StatePublished - Dec 1 2014

Fingerprint

Clustered Regularly Interspaced Short Palindromic Repeats
Cell Survival
Stem Cells
Genes
Heme Oxygenase-1
3' Untranslated Regions
Introns
Transcriptional Activation
Myocardium
Anti-Inflammatory Agents
Guide RNA
Zinc Fingers
Cell- and Tissue-Based Therapy
MicroRNAs
Myocardial Ischemia
Cell Death
Complementary DNA
Transplantation
Genome
Gene Expression

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Enhancing stem cell survival in an ischemic heart by CRISPR-dCas9-based gene regulation. / Pan, Alexander; Weintraub, Neal Lee; Tang, Yao Liang.

In: Medical Hypotheses, Vol. 83, No. 6, 01.12.2014, p. 702-705.

Research output: Contribution to journalArticle

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