Development of a Simultaneous Knock-Down/Replacement Vector for Human Cells
Development of gene-exchange technology.
Project is Not Currently Available
This project has reached full capacity for the current term. Please check back next semester for updates.
Project description
While CRISPR and siRNA technologies allow for the knock-out and knock-down of genes in human cells for studying their functions, these technologies are limited in studying genes essential for human cells. This project aims to address this by developing a single vector with two cassettes: one for insertion of of sequences designed to create an shRNA for knock-down of expression of the natural copy of the essential gene, and a second cassette that encodes a recombinant copy of the essential gene designed to not be targeted by the shRNA sequence. This single vector will allow researchers to carry out a rapid "exchange" of the normal cellular essential gene with a recombinant version of the gene. This vector will allow researchers working on essential cellular genes to rapidly analyze function of various mutations of those genes in cultured human cells, greatly facilitating research on essential cellular factors.
Project outcome
The outcome of this project will be to develop this vector, and using the test protein sequences (the essential RPA2 gene and a known functional recombinant form of that gene, GFP-RPA2) show that upon transfection of human cell lines over time the native RPA2 protein levels decrease while the GFP-RPA2 protein levels increase.
Project details
| Length of commitment | Longer than a semester (about 6-9 months) |
| Start time | Spring |
| In-person, remote, or hybrid? | In-person |
| Level of collaboration | Individual student project |
| Benefits | Stipend Potential academic credit (through BCH498) |
| Who is eligible | Freshmen, sophomores, and juniors who have taken CHE101, CHE113, BIO201, BIO201Lab, CHE205Lab, MIC401. |
Core partners
- Dr. Rama Dey-Rao
- Dr. Thomas Melendy
Project mentor
Thomas Melendy
Associate Professor
Microbiology & Immunology
Phone: (716) 829-3789
Email: TMelendy@buffalo.edu
Start the project
- 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.)
- 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.)
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. After you’re approved to begin the project, your mentor will send the relevant materials. Please reference this when you get to Step 2 of the Preparation Phase.
Write a 2-to 3-page analysis of the pros and cons of various methods for knocking-out or knocking-down expression of specific proteins in cultured human cells.
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Keywords
microbiology, immunology, biology, chemistry, genes, CRISPR