Molecular Biology and Gene Delivery Core

University Of California Los Angeles, Los Angeles CA

Investigators

Linked publications & trials

Abstract

PROJECT SUMMARY – MBGD CORE The Molecular Biology & Gene Delivery (MBGD) core is a consolidated core of the previous Biochemistry and Molecular Instrumentation (BMBI) core and Gene & Cell Delivery (G&CD) core. This MBGD core is an essential resource supporting vision research activities at UCLA and will be heavily used in the coming years. The director of the MBGD core is Dr. Xian-Jie Yang, Professor of Ophthalmology, with expertise in molecular biology, neurodevelopment, stem cell biology, mouse genetics, and viral vector-mediated gene delivery. The major part of the MBGD core consists of common equipment and instrumentations required for research using biochemistry and molecular biology approaches. These include instruments such as autoclaves, various spectrophotometers, centrifuges, incubators, microplate readers, immunoassay protein analyzers, and PCR machines for DNA amplification and RNA analysis. Most of these instruments are basic but their utilization is extensive and fundamental for the specific research program of each laboratory. A majority of our core investigators employ basic biochemistry and molecular biological tools to prepare and analyze various biological samples qualitatively and quantitatively. The research activities include molecular cloning, RNA and DNA preparation, PCR genotyping of genetically engineered mice, gene expression analysis by qPCR, mutagenesis and CRISPR-Cas gene editing, protein extraction and expression analysis using Western blot, biochemical and enzymatic assays by microplate readers. Besides the common instruments, this core also includes specialized spectrophotometers with capabilities of fluorescence, UV and near/infra-red wavelengths. Several recently acquired instruments, including the Lonza 4d-Nucleofector, Applied Biosystems Digital Absolute Q for digital PCR, and SONY MA900 Multi-Application Cell Sorter, have further updated and enhanced the capacity of the MBGD core. To serve the research needs of gene delivery, the MBGD core has established a viral production facility to produce recombinant lentiviruses and AAVs for UCLA vision researchers. This facility is staffed with a specialist who is skilled to produce high titer purified lentiviral and AAV stocks at low cost for various research projects involving gene delivery in vitro and in vivo. The MBGD core director Dr. Xian-Jie Yang will consult with investigators on viral vector designs and supervise the core specialist to produce viral stocks and perform quality control assays. The MBGD core also has a designated mouse room, which is equipped with micromanipulators and a square-wave generator for in vivo eye electroporation and intraocular injections. The same facility can house post-operative mice deemed to need BSL2 containment. The MBGD core director Dr. Xian-Jie Yang will supervise the viral production and the core technician to ensure the continuous functionality and reliability. Additionally, each laboratory has a senior staff researcher who provides support and distributes established protocols to new trainees in our research community.