Novel In Vitro Selection of <i>Trans</i>-Acting <i>BCL-2</i> mRNA-Cleaving Deoxyribozymes for Cancer Therapy
The B Cell Lymphoma-2 (<i>BCL-2</i>) family proteins are central regulators of apoptosis, and their dysregulation is frequently associated with cancer progression and resistance to therapy. While small molecules like venetoclax have shown promise, nucleic acid-based therapeutics targetin...
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MDPI AG
2025-06-01
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| author | Veera Vijaya Basamshetty Vijay Kumar Gangipangi Uppulapu Shravan Kumar Santhosh Shanthi Bhupathi Sridhar Reddy Kaulagari Prashant Giri Swapnil Sinha Utpal Mohan Konstantinos Sdrimas |
| author_facet | Veera Vijaya Basamshetty Vijay Kumar Gangipangi Uppulapu Shravan Kumar Santhosh Shanthi Bhupathi Sridhar Reddy Kaulagari Prashant Giri Swapnil Sinha Utpal Mohan Konstantinos Sdrimas |
| author_sort | Veera Vijaya Basamshetty |
| collection | DOAJ |
| description | The B Cell Lymphoma-2 (<i>BCL-2</i>) family proteins are central regulators of apoptosis, and their dysregulation is frequently associated with cancer progression and resistance to therapy. While small molecules like venetoclax have shown promise, nucleic acid-based therapeutics targeting <i>BCL-2</i> remain underexplored. Here, we report a novel in vitro evolution strategy to generate trans-acting RNA-cleaving DNAzymes targeting natural <i>BCL-2</i> mRNA without requiring covalent substrate-linking. Using a 50-base region of <i>BCL-2</i> mRNA as a selection target, we evolved several DNAzymes that demonstrate significant RNA cleavage activity. These DNAzymes downregulated <i>BCL-2</i> expression, induced apoptosis, and reduced cell viability in HepG2 and MCF-7 cancer cells. In vivo, our novel DNAzymes significantly suppressed tumor growth in a syngeneic mouse breast cancer model, with efficacy comparable to 5-Fluorouracil. This study presents a proof of concept for a novel strategy to evolve functional DNAzymes against native mRNA sequences and highlights their potential as gene-silencing tools in cancer therapy. Future studies will explore the therapeutic potential of these findings in cancer patients. Additionally, investigating the underlying molecular mechanisms in more complex cancer models will further validate the observed effects. |
| format | Article |
| id | doaj-art-596ef94aa6bf4d40b7128cd93a8ebc9c |
| institution | OA Journals |
| issn | 2073-4409 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Cells |
| spelling | doaj-art-596ef94aa6bf4d40b7128cd93a8ebc9c2025-08-20T02:35:59ZengMDPI AGCells2073-44092025-06-01141394510.3390/cells14130945Novel In Vitro Selection of <i>Trans</i>-Acting <i>BCL-2</i> mRNA-Cleaving Deoxyribozymes for Cancer TherapyVeera Vijaya Basamshetty0Vijay Kumar Gangipangi1Uppulapu Shravan Kumar2Santhosh Shanthi Bhupathi3Sridhar Reddy Kaulagari4Prashant Giri5Swapnil Sinha6Utpal Mohan7Konstantinos Sdrimas8Department of Medical Oncology, West Virginia Cancer Institute, West Virginia University, Morgantown, WV 26505, USADepartment of Pathology and Laboratory Medicine, Lewis Katz School of Medicine, Temple University, Philadelphia, PA 19140, USADepartment of Internal Medicine Carver College of Medicine University of Iowa, Iowa City, IA 52242, USADepartment of Pharmaceutical Sciences, West Virginia University, Morgantown, WV 26505, USADepartment of Pharmaceutical Sciences, West Virginia University, Morgantown, WV 26505, USADepartment of Cell Stress Biology, Roswell Park Comprehensive Cancer Center, Buffalo, NY 14263, USAIIT Guwahati TIDF BioNest, Guwahati 70005, IndiaNational Institute of Pharmaceutical Education and Research, Kolkata 70005, IndiaDepartment of Medical Oncology, West Virginia Cancer Institute, West Virginia University, Morgantown, WV 26505, USAThe B Cell Lymphoma-2 (<i>BCL-2</i>) family proteins are central regulators of apoptosis, and their dysregulation is frequently associated with cancer progression and resistance to therapy. While small molecules like venetoclax have shown promise, nucleic acid-based therapeutics targeting <i>BCL-2</i> remain underexplored. Here, we report a novel in vitro evolution strategy to generate trans-acting RNA-cleaving DNAzymes targeting natural <i>BCL-2</i> mRNA without requiring covalent substrate-linking. Using a 50-base region of <i>BCL-2</i> mRNA as a selection target, we evolved several DNAzymes that demonstrate significant RNA cleavage activity. These DNAzymes downregulated <i>BCL-2</i> expression, induced apoptosis, and reduced cell viability in HepG2 and MCF-7 cancer cells. In vivo, our novel DNAzymes significantly suppressed tumor growth in a syngeneic mouse breast cancer model, with efficacy comparable to 5-Fluorouracil. This study presents a proof of concept for a novel strategy to evolve functional DNAzymes against native mRNA sequences and highlights their potential as gene-silencing tools in cancer therapy. Future studies will explore the therapeutic potential of these findings in cancer patients. Additionally, investigating the underlying molecular mechanisms in more complex cancer models will further validate the observed effects.https://www.mdpi.com/2073-4409/14/13/945DNAzymes<i>BCL-2</i>apoptosiscancersynthetic biology |
| spellingShingle | Veera Vijaya Basamshetty Vijay Kumar Gangipangi Uppulapu Shravan Kumar Santhosh Shanthi Bhupathi Sridhar Reddy Kaulagari Prashant Giri Swapnil Sinha Utpal Mohan Konstantinos Sdrimas Novel In Vitro Selection of <i>Trans</i>-Acting <i>BCL-2</i> mRNA-Cleaving Deoxyribozymes for Cancer Therapy Cells DNAzymes <i>BCL-2</i> apoptosis cancer synthetic biology |
| title | Novel In Vitro Selection of <i>Trans</i>-Acting <i>BCL-2</i> mRNA-Cleaving Deoxyribozymes for Cancer Therapy |
| title_full | Novel In Vitro Selection of <i>Trans</i>-Acting <i>BCL-2</i> mRNA-Cleaving Deoxyribozymes for Cancer Therapy |
| title_fullStr | Novel In Vitro Selection of <i>Trans</i>-Acting <i>BCL-2</i> mRNA-Cleaving Deoxyribozymes for Cancer Therapy |
| title_full_unstemmed | Novel In Vitro Selection of <i>Trans</i>-Acting <i>BCL-2</i> mRNA-Cleaving Deoxyribozymes for Cancer Therapy |
| title_short | Novel In Vitro Selection of <i>Trans</i>-Acting <i>BCL-2</i> mRNA-Cleaving Deoxyribozymes for Cancer Therapy |
| title_sort | novel in vitro selection of i trans i acting i bcl 2 i mrna cleaving deoxyribozymes for cancer therapy |
| topic | DNAzymes <i>BCL-2</i> apoptosis cancer synthetic biology |
| url | https://www.mdpi.com/2073-4409/14/13/945 |
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