**HONORABLE MENTION**
Student(s): Jose–Gabriel Castro
Project: Functionalized Cell Penetrating Peptides for the Development of Targets CRISPR Delivery System | View Poster (PDF)
Major(s): Bioengineering
Advisor(s): Tomas Gonzalez-Fernandez
Abstract
Tissue engineering has allowed for the development of a variety of therapies targeting different diseases and non-healing injuries. To address the need for more specific treatment targets, clustered regularly interspaced short palindromic repeat (CRISPR) based therapies are precise and effective for gene editing applications. Viral vectors and lipid-based nanoparticles are commonly used delivery systems for gene editing but due to drawbacks such as cytotoxic effects and cargo size limits, extensive research has been conducted aiming to develop a suitable vector that overcomes these limitations. Cell penetrating peptides, such as the arginine-alanine-leucine-alanine (RALA) peptide, can deliver diverse cargo types while preserving cell viability and proliferation. In fact, RALA has previously delivered multiple CRISPR cargoes to achieve CRISPR-Knock In (CRISPR-KI), CRISPR-Knock Out (CRISPR-KO) and CRISPR activation (CRISPRa) in primary stem cells. Although RALA can be harnessed for the efficient delivery of CRISPR components, RALA is not cell specific which hinders its safety and efficiency. To address these limitations, we modified the RALA peptide sequence to limit its CRISPR delivery capabilities to primary mesenchymal stromal cells (MSCs) through the conjugation of an MSC targeting peptide onto the RALA sequence. The integration of the MSC targeting peptide not only allowed for a degree of cell specificity but also increased the cell viability and yield of transfected cells, leading to a higher level of clinical relevance for this novel CRISPR delivery system.
About Jose–Gabriel Castro
Jose-Gabriel is a current 3rd year bioengineering student with a focus on biomaterials and tissue engineering. Having started in the TGF lab in early 2023, he aims to functionalize a cell penetrating peptide to produce a cell-specific transfection vector for clinically relevant CRISPR therapeutics in primary stem cells. In the summer of 2024, he conducted research focusing on the viral transduction of alginate-encapsulated induced pluripotent stem cells for advanced microscopic visualization at the University of Bordeaux through the Iacocca International Internship Program. After graduation, he is planning to continue his education through graduate studies and specialize in applying synthetic biology tools for neural engineering applications. Along with the lab, he is on the executive board for Lehigh’s chapter of the Biomedical Engineering Society (BMES), president of the campus’ Reformed University Fellowship (RUF) and volunteers through several opportunities around Lehigh.