Chemicals that pass through hereditary materials to disrupt hormone systems

Problem Title

Chemicals that pass through hereditary materials to disrupt hormone systems

Scientific Title

A rapid method for stabilization and isolation of high quality sperm nucleic acids

Student: 
Gianna Luchetti
Major(s): 
Public Health Sciences
iCons Concentration: 
Biomedicine/Biosystems
iCons Class Year: 
Class of 2014
Executive Summary 

There is a growing interest in determining the role of sperm genetics and epigenetic changes on reproductive success and the life-course trajectory of health outcomes. Animal models have shown that RNA and epigenomic changes to a sperm cell can impact both reproductive success and offspring health outcomes.

The greatest current challenge in studying sperm epigenetics as well as RNA in humans is a lack of a practical method for storage and extraction. Human samples for biological research are often difficult to acquire, and limited in quantity, making a improved method necessary for optimal research. Current methods include long incubation times, odorous chemicals, a lack of stabilization, and low quantities of DNA and RNA extracted. In addition sperm samples have much lower concentrations of RNA as compared to somatic cells making proper storage and processing important.

Exploring current storage and extraction methods and commercially available kits, we produced a rapid method for stabilization and isolation of high quality sperm nucleic acids using commercially available reagents that are stable at room temperature. The use of TCEP, a strong odorless guanidine-based lysis method, along with proteinase K has expedited the process of extracting stable nucleic acids and is safe in clinical laboratories. The sperm lysate can be then be processed using a commercial kit for either DNA or RNA with comparable yields to previous protocols which require longer incubations. The applications of this new method will significantly increase the ability to practically carry out research on the sperm epigenome as well as RNA.

Problem Keywords: 
hormones
Scientific Keywords: 
endocrine disruptors
sperm epigenome
TCEP