Silencing of Cav1.2 gene in neonatal cardiomyocytes by lentiviral delivered shRNA

Eddy Karnabi, Yongxia Qu, Salvatore Mancarella, Yuankun Yue, Raj Wadgaonkar, Mohamed Boutjdir

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Cav1.2 (α1C) and Cav1.3 (α1D) L-type Ca channels are co-expressed in the heart. To date, there are no pharmacological or biophysical tools to separate α1D from α1C Ca currents (ICa-L) in cardiomyocytes. Here, we established a physiological model to study α1D ICa-L in native myocytes using RNA interference. Transfection of rat neonatal cardiomyocytes (RNC) with α1C specific siRNA resulted in low silencing efficiency (50-60%) at the mRNA and protein levels. The use of lentivirus shRNA resulted in 100% transfection efficiency and 92% silencing of the α1C gene by real-time PCR and Western blot. Electrophysiological experiments showed that the total ICa-L was similarly reduced by 80% in lentivirus transfected cells. Both biochemical and functional data demonstrated high transfection and silencing efficiency in the cardiomyocytes using lentiviral shRNA. This novel approach allows for the assessments of the roles of α1C and α1D Ca channels in native myocytes and could be used to examine their roles in physiological and pathological settings.

Original languageEnglish (US)
Pages (from-to)409-414
Number of pages6
JournalBiochemical and Biophysical Research Communications
Volume384
Issue number4
DOIs
StatePublished - Jul 10 2009

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Gene Silencing
Cardiac Myocytes
Small Interfering RNA
Transfection
Lentivirus
Genes
Muscle Cells
Physiological models
RNA Interference
Rats
Real-Time Polymerase Chain Reaction
Western Blotting
Pharmacology
RNA
Messenger RNA
Proteins
Experiments

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Biophysics
  • Cell Biology
  • Molecular Biology

Cite this

Silencing of Cav1.2 gene in neonatal cardiomyocytes by lentiviral delivered shRNA. / Karnabi, Eddy; Qu, Yongxia; Mancarella, Salvatore; Yue, Yuankun; Wadgaonkar, Raj; Boutjdir, Mohamed.

In: Biochemical and Biophysical Research Communications, Vol. 384, No. 4, 10.07.2009, p. 409-414.

Research output: Contribution to journalArticle

Karnabi, Eddy ; Qu, Yongxia ; Mancarella, Salvatore ; Yue, Yuankun ; Wadgaonkar, Raj ; Boutjdir, Mohamed. / Silencing of Cav1.2 gene in neonatal cardiomyocytes by lentiviral delivered shRNA. In: Biochemical and Biophysical Research Communications. 2009 ; Vol. 384, No. 4. pp. 409-414.
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