Generation of cell-type specific ATF6 knock-in and knock-out double transgenic lines, and spatial analysis of their lungs and hearts

Cellular Symphony: Harmonious Collaboration and the Impact of Environmental Toxins.

Cigarette smoke-induced proteostatsis disruption: implications for aging lungs and hearts. This project investigates how sustained ATF6 activation contributes to epithelial and vascular dysfunction in the lungs and hearts of smokers. 

Project description

Student will work under close supervision from a graduate student in the lab on the project that investigates how cigarette smoking causes lung and heart pathologies. Student will learn how to handle animals, and how to breed Cre-driver mouse strain with floxed alleles of the genes of interest (knock-in and knock-out alleles). Student is expected to understand and will be taught the theory behind the set-up of the breeding pairs for knock-in and knock-out alleles. Student will learn genotyping techniques of (F1) animals and identification of double transgenic animals. Student also will learn how to induce Cre-expression and validate desired effects., how to expose animals to cigarette smoke, and how to harvest their lungs and hearts. Student will understand lungs and hearts embedding techniques and learn slide preparation from paraffin blocks using microtome. Student will further learn how to evaluate and validate different antibodies by immunofluorescence, and how to identify proteins of interest in harvested lungs and hearts. Validated antibodies will be conjugated to metals (not by a student, but student may shadow the conjugation and learn the principles of the technique) and used in imaging mass cytometry to evaluate spatial changes in different cell types caused by smoking. If time allows, the student will work closely with a graduate student partner, and learn imaging mass cytometry, and data analysis. Research on this project requires efficiency, time planning and strong teamwork. Having credits in the sciences is a prerequisite. Biology, cell biology, physiology and some python understandings are welcome. 

Project outcome

Student is expected to learn animals handling, genotyping, immunofluorescence of harvested and embedded tissues and be introduced to imaging mass cytomerty. By the end of the program, the student is expected to be able to plan their own experiments, design and include appropriate controls, conclude if the antibody staining is specific, be aware of the consideration for the size of the groups for analysis, as well as of the statistical analysis required to validate conclusions of the experiment. Student is also expected to learn to work as a team, and generate or/and contribute to generation of the figures for the a future publication. Students may be asked to write methods part and to work on the figure. If student desires, they will be welcome to participate in the department research day, along with their graduate student partner. In this case, students will also have an introductory experience to the abstract writing and poster presentation. Therefore, this project will serve as a gateway to a better student scientific emergence, provide an opportunity to learn new skills and to develop scientific thinking in the professional scientific community. The ideal end goal is to give a student mentored experience where they can further develop their laboratory skills at any levels, and possibly co-author the manuscript. 

Project details

Timing, eligibility and other details
Length of commitment Year-long (10-12 months)
Start time Summer (May/June of 2025)
In-person, remote, or hybrid? In-Person Project (Can only function with in-person engagement)
Level of collaboration Small group project (2-3 students)
Benefits

Academic Credit

Stipend

Who is eligible Seniors with an understanding of animal handling and principles of immunihistochemistry; ability to work in R. Biology, cell biology, physiology and some python understandings are welcome. 

Project mentor

Anna Blumental-Perry

Assistant Professor

Biochemistry

Phone: (716) 829-2105

Email: annablum@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.) 

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. 

  • Student should read about the project, and understand the basic concept of mouse genetics: Punnett's square, two genes distribution, cell- type specific Cre drivers, LoxP sites, Rosa26 locus, knock-in and knock-out alleles.
  • Students need to read review and understand the idea of Unfolded protein response pathway.
  • Student will need to do animals training and be added to the animals protocols active in the lab. 

Keywords

medicine, transgenic mice, spatial transcriptomics, Biochemistry, Jacobs School of Medicine and Biomedical Sciences