# Administrative Equipment Supplement to Genetics and Molecular Biology of Striated Muscle Myosin

> **NIH NIH R37** · SAN DIEGO STATE UNIVERSITY · 2023 · $35,289

## Abstract

ABSTRACT
This equipment supplement requests funds to purchase an Applied Biosystems QuantStudio 3 Real-Time PCR
System, which is critical for completing the aims of the parent grant (NIH MERIT award to Sanford I. Bernstein
entitled Genetics and Molecular Biology of Striated Muscle Myosin, GM032443). This instrument will replace an
unserviceable Applied Biosystems model 7000, which is approximately 20 years old and is no longer being
calibrated or repaired by the vendor. Our current instrument is providing inconsistent results and has shut down
prematurely during operation. The QuantStudio 3 Real-Time PCR System will be essential for completing our
study on roles of aging-related aggregating proteins in myosin-based skeletal muscle diseases. In this aim, we
are assessing the functions of proteins that appear at abnormal levels in aggregates affiliated with Drosophila
models of inclusion body myopathy type 3 or myosin storage myopathy. This is accomplished by knocking down
(RNAi) or over-expressing (UAS-Gal4 transgenesis) each of these identified proteins in wild-type adults and
assessing muscle structure and function. This transcript manipulation approach is then used in the disease
model backgrounds to assess possible improvement in muscle phenotypes. For these studies, it is essential to
employ real-time quantitative PCR to determine the levels of over- or under-expression of each abnormally
expressed gene in both the disease models and in the up- or down-regulated backgrounds. This approach is
also useful in assessing transgene expression for our other two grant aims, where we are transgenically
increasing levels of the FOXO transcription factor (which improves proteostasis by enhancing clearance of
aggregates), to assess whether structural and functional improvements occur in a) skeletal muscles that contain
aging-related post-translational myosin modification mimics or b) cardiac muscles that serve as models of dilated
cardiomyopathy. The QuantStudio PCR System is advantageous compared to our current outmoded and poorly
functional instrument in that it requires less frequent calibration, has VeriFlex temperature control technology to
optimize PCR conditions, has better detection capability and resolution, and has onboard and cloud data storage.
In addition to these instrument advantages, we will be moving from SYBR Green to TaqMan qPCR assays,
wherein we will not simply detect amplified double-stranded DNA (including non-specific amplifications) but will
only detect gene-specific amplification. The TaqMan system also has the availability of a library of over 40,000
verified Drosophila primers/probes and the ability to perform multiplex amplification of both the experimental and
control transcripts in a single tube, thereby enhancing efficiency of probe design and reproducibility of
experimental conditions. Overall, the QuantStudio 3 Real-Time PCR System will provide efficiency, reliability
and rigor that is lacking with our outmoded and unrel...

## Key facts

- **NIH application ID:** 10772908
- **Project number:** 3R37GM032443-38S2
- **Recipient organization:** SAN DIEGO STATE UNIVERSITY
- **Principal Investigator:** Sanford I Bernstein
- **Activity code:** R37 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2023
- **Award amount:** $35,289
- **Award type:** 3
- **Project period:** 1983-07-01 → 2027-04-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10772908

## Citation

> US National Institutes of Health, RePORTER application 10772908, Administrative Equipment Supplement to Genetics and Molecular Biology of Striated Muscle Myosin (3R37GM032443-38S2). Retrieved via AI Analytics 2026-05-24 from https://api.ai-analytics.org/grant/nih/10772908. Licensed CC0.

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