Development of a Transient Gene Replacement System for Human Cells

Dr. Melendy's laboratory carries out basic molecular and cellular biology of human cells, particularly with relevance to viruses and cancer.

Develop a system for cultured human cells to carry out transient protein swap for essential cellular proteins. 

Project description

This project will be to develop a cloning vector for human cells that will allow the user to express a shRNA to downregulate an essential cellular gene while simultaneously expressing another copy of that gene that is resistant to downregulation. The vector will be designed in consultation with other laboratory members. The vector will be subjected to sequencing to verify the work in constructing the vector. Finally the student will carry out transfections, selection, and induction of the shRNA, and immunoblotting to ascertain whether the desired protein switch has occurred. 

Project outcome

Project outcome will be a human cell protein exchange vector, and analysis of function in human cells. 

Project details

Timing, eligibility and other details
Length of commitment About a semester; 3-5 months
Start time

Summer (May/June) 

Fall (August/September)

  
In-person, remote, or hybrid? In-Person Project 
Level of collaboration Individual student project 
Benefits Academic credit
Stipend 
Who is eligible Sophomores & Juniors who have basic molecular biology laboratory skills and knowledge in DNA vector design and construction, human cell transfection, immunoblotting 
 

Project mentor

Thomas Melendy

Associate Professor

Microbiology & Immunology

Phone: (716) 829-3789

Email: TMelendy@buffalo.edu

Start the project

  1. Email the project mentor using the contact information above to express your interest and get approval to work on the project. (Here are helpful tips on how to contact a project mentor.)
  2. After you receive approval from the mentor to start this project, click the button to start the digital badge. (Learn more about ELN's digital badge options.) 
Start the Digital Badge

Preparation activities

Once you begin the digital badge series, you will have access to all the necessary activities and instructions. Your mentor has indicated they would like you to also complete the specific preparation activities below. Please reference this when you get to Step 2 of the Preparation Phase. 

Plasmid vector design. Plasmid vector construction and sequencing for verification. Transfection of cultured human cells. Immunoblot analysis to analysis effectiveness. 

Keywords

Transient gene replacement, Microbiology, Biochemistry