ساخت اکانت کاربری رایگان

مقالات پذیرفته شده کنگره

  • صفحه نخست
  • مقالات پذیرفته شده اولین کنگره بین المللی ژنتیک سرطان
بازنشر

  • CRISPR Models for Gene Knockout in Transcription-Coupled Repair and DNA-Protein Crosslink Outcomes in Lymphoid Leukemia

  • Neda Zahmatkesh,1,*
    1. Msc of Molecular Genetic Department of Genetics, Zanjan Branch, Islamic Azad University, Zanjan, Iran.


  • Introduction: Lymphoid leukemias, which include both acute lymphoblastic leukemia (ALL) and chronic lymphocytic leukemia (CLL), are characterized by a range of genetic and epigenetic abnormalities that drive their initiation and progression. Among the various forms of DNA damage observed in these cancers, DNA-protein crosslinks (DPCs) stand out as particularly harmful. These bulky lesions disrupt essential cellular processes like transcription and replication, posing a significant threat to genome stability. Transcription-coupled repair (TCR), a specialized sub-pathway of nucleotide excision repair, is responsible for the rapid removal of lesions that block transcription on the active strand of genes. TCR plays a central role in resolving DPCs and maintaining genomic integrity in rapidly dividing cells like those in leukemia. Recent advances in CRISPR-Cas9 gene-editing technology have enabled the creation of highly specific knockout models, allowing researchers to investigate the functional importance of TCR-related genes and their role in DPC repair in leukemia. This review summarizes current knowledge derived from CRISPR-based models, shedding light on the molecular mechanisms underlying TCR and DPC response in lymphoid leukemia and their potential as therapeutic targets.
  • Methods: A systematic literature review was conducted using databases such as PubMed, Scopus, and Web of Science, focusing on studies published between 2018 and 2024. Search terms included “CRISPR,” “transcription-coupled repair,” “DNA-protein crosslink,” “lymphoid leukemia,” “gene knockout,” and “genome editing.” The selected studies included both in vitro and in vivo research that employed CRISPR to disrupt genes involved in the TCR pathway within the context of lymphoid malignancies. The findings were critically analyzed to integrate mechanistic insights with their therapeutic relevance.
  • Results: CRISPR-Cas9 genome editing has proven to be a powerful tool for dissecting the roles of key TCR genes such as CSB (ERCC6), CSA (ERCC8), and SPRTN in lymphoid leukemia models. Knocking out these genes in leukemia cell lines leads to defective DPC repair, resulting in increased DNA damage, transcriptional arrest, and heightened cell death—clearly highlighting their vital role in preserving genomic integrity. For example, SPRTN, a protease involved in the proteolytic degradation of crosslinked proteins, is essential for DPC resolution. Its deletion sensitizes leukemia cells to chemotherapeutic agents like topoisomerase inhibitors, which induce DPCs as part of their mechanism of action. Similarly, disruption of CSA and CSB impairs the recruitment and function of the nucleotide excision repair machinery, further exacerbating DNA damage and reducing cell viability. CRISPR-generated models have also uncovered compensatory interactions between TCR and other DNA repair pathways, such as homologous recombination, indicating a tightly regulated network of DNA damage response mechanisms in leukemic cells. Furthermore, high-throughput CRISPR screens have identified new modulators of TCR efficiency and DPC tolerance, including factors involved in ubiquitination, chromatin remodeling, and protein degradation. These findings point to the potential of combination therapies that target multiple DNA repair mechanisms simultaneously, offering promising strategies to overcome chemoresistance in lymphoid leukemia.
  • Conclusion: CRISPR-Cas9 technology has dramatically advanced our understanding of transcription-coupled repair and its role in managing DNA-protein crosslink damage in lymphoid leukemias. Gene knockout studies have highlighted the critical importance of TCR components in maintaining transcriptional integrity and genome stability, particularly in the face of genotoxic stress. These insights not only enhance our fundamental understanding of leukemia biology but also pave the way for identifying novel therapeutic targets and predictive biomarkers for treatment response. Future research should prioritize expanding CRISPR-based screening approaches and developing more sophisticated in vivo models to capture the complexity of DNA repair dynamics. Such efforts will be essential in designing effective, targeted therapies that sensitize leukemia cells to both conventional and emerging chemotherapeutic agents.
  • Keywords: CRISPR-Cas9, Transcription-coupled repair, DNA-protein crosslinks, Gene knockout, Lymphoid leukemia

به خانواده بزرگ کنسر ژنتیکس و ژنومیکس سرطان بپیوندید!

ایجاد حساب کاربری رایگان