Viral Gene Delivery in the Mouse Heart for Evaluating Gene Effects on Heart Function

Abstract

Source: Lu, A., et al. Viral Transgene Expression in Rodent Hearts and the Assessment of Cardiac Arrhythmia Risk. J. Vis. Exp. (2022)

The video demonstrates a method for ultrasound-guided intramyocardial injection of a recombinant viral vector in a mouse model. This technique enables targeted gene delivery to the heart for studying the effects of transgene expression on cardiac function.

Protocol

1. Viral transgene expression in rodent ventricular tissues

NOTE: Store recombinant adenovirus or adeno-associated virus (AAV) that expresses the target gene and the corresponding control virus, such as Ad-GFP (Green fluorescent protein) (titer of 1 x 10¹⁰ plaque-forming units [PFU]/mL) or AAV9-GFP (titer of 1 x 10¹³ genome copies [GC]/mL) (see Table of Materials), in a −80 °C freezer.

1. On the day of virus injection, thaw the virus on ice. Aspirate the virus expressing the target gene and the control virus into two separate syringes (volume = 50 µL) with 30 G ½ needles and....

Materials

List of materials used in this article

Name	Company	Catalog Number	Comments
30 G ½ PrecisionGlide Needle	Becton Dickinson (BD)	305106
Adeno-associated virus (Green flourescent protein(GFP) expressing AAV9)	Vector Biolabs	7007
Adenovirus (Ad-GFP)	Vector Biolabs	1060
Biosafety cabinet (Level II)	Microzone Corporation	N/A	Model #: BK-2-4
Buprenorphine	Vetergesic	DIN 02342510
Hair clipper	WAHL Clipper Corporation	78001
Hamilton syringe	Sigma-Aldrich	20701	705 LT, volume 50 μL
Heating pad	Life Brand	E12107
Isoflurane	Fresenius Kabi Ltd.	M60303
Mice (Ctnnb1flox/flox)	Jackson Labs	4152
Mice (αMHC-MerCreMer)	Jackson Labs	5650
Ophthalmic ointment	Systane	DIN 02444062
Rat (Sprague-Dawley, male)	Charles River	400
Scalpel blades	Fine Science Tools	10010-00
Scalpel handle	Fine Science Tools	10007-12
VEVO3100 Preclinical Imaging System	VisualSonic Inc.	N/A