Beta-thalassemia is a genetic disorder that significantly burdens healthcare systems globally. This inherited blood disorder, categorized into beta-thalassemia and alpha-thalassemia, results in insufficient globin production, leading to anemia and iron overload from frequent transfusions. Severe cases, known as thalassemia major, require regular blood transfusions. Beyond clinical suspicion and biochemical tests, molecular techniques are essential for confirming the diagnosis and guiding treatment. Advanced molecular profiling methods such as Polymerase Chain Reaction (PCR), Multiplex Ligation-dependent Probe Amplification (MLPA), Next-Generation Sequencing (NGS), Third-Generation Sequencing (TGS), and Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) are effective in detecting mutations. Epigenetic factors also play a crucial role, driving the development of epidrugs for targeted therapy. This review covers various molecular techniques, established gene-editing methods, epigenetic mechanisms, and the impact of artificial intelligence on thalassemia management. It highlights the importance of selecting precise and sensitive molecular tools for detecting thalassemia gene mutations and stresses the need to make these testing methods accessible in resource-limited clinical settings.

Background: Germline mutations in high-penetrance genes contribute to the development of colorectal cancer with a >1% familial risk. The proband with a family history of cancer was at an increased risk for colorectal cancer development. However, data on the germline mutational landscape in non-Western populations, particularly in India, remain limited. Objective: This study aimed to investigate the prevalence of germline mutations in individuals fulfilling clinical criteria for suspected Lynch Syndrome (LS) or familial adenomatous polyposis (FAP) using a targeted multigene next-generation sequencing (NGS) approach. Method: Mutation analysis was performed for 110 suspected Lynch syndrome and 12 familial adenomatous polyposis (FAP) and attenuated FAP (AFAP) patients. Sanger sequencing was employed to validate pathogenic, likely pathogenic, and novel variants. Results: Pathogenic mutations were identified in 20% of suspected Lynch syndrome cases and 66% of FAP cases, indicating a stronger genetic contribution in FAP. Notably, this is the first study to report a FANCD2 mutation in FAP. Most LS-associated mutations occurred in mismatch repair genes ( MLH1, MSH2, MSH6 ), while APC and MUTYH mutations predominated in FAP. Four novel mutations were identified: MLH1 c.676delC, MLH1 c.378delC, MDC1 c.2839C>T , and RAD52 c.1215delG . Functional prediction of 100 VUS revealed several potentially disruptive variants, notably BLM P866L , which showed significant loss of protein–protein interactions. A pathogenic FANCD2 mutation was identified in one FAP family, suggesting a broader spectrum of contributing genes. Conclusion: This study demonstrates the utility of multigene NGS panels in detecting clinically actionable germline mutations in suspected hereditary CRC cases in an Indian population. The identification of novel and ethnically distinct variants highlights the need for population-specific genetic databases and reinforces the role of genetic screening and counseling in cancer prevention and early diagnosis.

Haematological malignancies comprise a diverse group of life-threatening systemic diseases in the blood, bone marrow, and lymphoid tissues, and their progression is strongly influenced by a supportive tumour microenvironment that drives proliferation and drug resistance. Drug development and personalised treatment using patient-derived cells are a tedious and expensive process, often conducted in 2D cell culture, which has many uncertainties and limitations due to poor mimicry of the microenvironment, cellular mechanisms, and morphological changes. Animal models also fail to fully replicate human pathobiology and are time-consuming. Alternatively, three-dimensional (3D) in vitro tissue modelling techniques can replicate the complex microenvironment, resembling the morphology, cell-cell interactions, and extracellular matrix (ECM) through methods like organoids, spheroids, tumoroids, and scaffold-based cultures. High-throughput drug screening in edited 3D cultures allows analysis of tumour interactions in haematological malignancies, for example, assessing gene-mediated drug resistance in AML within the marrow niche, depending on specific myeloma plasma cell cultures and their interactions with mesenchymal stem cells, along with susceptibility to CAR-T regulators and clinical translation in patient-derived therapies. The clinical translation of 3D culture and gene-editing–based in vitro models in leukemia, lymphoma, and multiple myeloma predicts cellular interactions with the tumour microenvironment, thereby enabling targeted treatment options for patients. This review highlights current progress in integrating gene editing with 3D culture system and emphasises their potential to enhance personalised drug discovery and preclinical testing in haematological malignancies.