Mutation In Exon Research Articles

Congenital Titinopathies Linked to Mutations in TTN Metatranscript-Only Exons

Congenital titinopathies reported to date show autosomal recessive inheritance and are caused by a variety of genomic variants, most of them located in metatranscript (MTT)-only exons. The aim of this study was to describe additional patients and establish robust genotype–phenotype associations in titinopathies. This study involved analyzing molecular, clinical, pathological, and muscle imaging features in 20 patients who had at least one pathogenic or likely pathogenic TTN variant in MTT-only exons, with onset occurring antenatally or in the early postnatal stages. The 20 patients with recessive inheritance exhibited a heterogeneous range of phenotypes. These included fetal lethality, progressive weakness, cardiac or respiratory complications, hyper-CKemia, or dystrophic muscle biopsies. MRI revealed variable abnormalities in different muscles. All patients presented severe congenital myopathy at birth, characterized by arthrogryposis (either multiplex or axial–distal) or neonatal hypotonia in most cases. This study provides detailed genotype–phenotype correlations in congenital titinopathies caused by mutations in MTT-only exons. The findings highlight the variability in clinical presentation and the severity of phenotypes associated with these specific genetic alterations. RNA-seq analyses provided valuable insights into the molecular consequences of TTN variants, particularly in relation to splicing defects and nonsense-mediated RNA decay. In conclusion, this study reinforces the genotype–phenotype correlations between congenital myopathies and variants in TTN MTT-only exons, improves their molecular diagnosis, and provides a better understanding of their pathophysiology.

DOCK8 gene mutation alters cell subsets, BCR signaling, and cell metabolism in B cells

DOCK8 deficiency has been shown to affect the migration, function, and survival of immune cells in innate and adaptive immune responses. The immunological mechanisms underlying autosomal recessive (AR) hyper-IgE syndrome (AR-HIES) caused by DOCK8 mutations remain unclear, leading to a lack of specific therapeutic options. In this study, we used CRISPR/Cas9 technology to develop a mouse model with a specific DOCK8 point mutation in exon 45 (c.5846C>A), which is observed in patients with AR-HIES. We then investigated the effect of this mutation on B cell development, cell metabolism, and function in a mouse model with Dock8 gene mutation. The results demonstrated that Dock8 gene mutation inhibited splenic MZ and GC B cell development and crippled BCR signaling. In addition, it resulted in enhanced glycolysis in B cells. Mechanistically, the reduced BCR signaling was related to decreased B cell spreading, BCR clustering, and signalosomes, mediated by inhibited activation of WASP. Furthermore, the DOCK8 mutation led to increased expression of c-Myc in B cells, which plays an important role in glycolysis. As such, GC B cells’ formation and immune responses were disturbed in LCMV-infected mice. These findings will provide new insights into the immunological pathogenesis of primary immunodeficiency disorder caused by DOCK8 mutation.

A single-base deletion in exon 2 of Hd1 delineates monogenic recessive photoperiod insensitivity in aromatic Joha rice: a novel allele for seasonal adaptability

BackgroundAssam's aromatic Joha rice is a unique rice class famous for its aroma, taste, and nutritional benefits, which fetch high market prices in domestic and international markets. Joha landraces are inherently poor yielders due to their strong aroma and predominantly photoperiod sensitivity. Hybridization involving non-aromatic HYVs improves yield with concomitant loss of quality. In this context, mutation breeding, a sustainable approach where genetic mutations are induced to create desirable traits, often provides useful allelic variation in specific morpho-agronomic traits. The present study delves into the genetic characterization of a photoperiod-insensitive mutant. As part of our mutation breeding programme, this mutant was isolated from a gamma ray-induced M2 population of a Joha rice landrace, Kon Joha.ResultsThe mutant was unique, and a single recessive gene conditions the induced photoperiod insensitivity. Mutant gene tagging involved 402 SSR and InDel markers, and later polymorphic markers were used for bulk segregant analysis (BSA) in the F2 population of ‘mutant × Kalijeera (distant parent)’. BSA revealed an association between the SSR marker RM527 and this mutant trait. This marker is present on chromosome 6 of the rice genome. Using chromosome 6-specific SSR markers in polymorphic screening and BSA revealed another associated marker, RM19725, for the mutant trait. The genomic interval between RM527 and RM19725 harbors a photoperiod-insensitive gene, Hd1, on chromosome 6. Cloning and sequencing of Hd1 genomic fragments from the parents and mutants revealed a single-base deletion in exon 2, leading to a frameshift mutation in the Hd1 protein. This mutation in exon 2 leads to severe structural abnormalities in the CCT domain of the Hd1 protein that is critical for the interaction of the repressing complex with conserved response elements in the florigen gene under long-day conditions, thereby causing photoperiod insensitivity.ConclusionsThe mutant's pleasant aroma and other quality characteristics, comparable to those of the parent cultivar, hold significant promise. They expand its potential use in a structured breeding programme aimed at developing high-value aromatic Joha rice. This rice, resilient to winter- and summer-growing environments and with broad seasonal adaptability, could revolutionize the rice market. The practical value of our research is underscored by this exciting possibility.

Short-Term Frequently Relapsing Ischemic Strokes Followed by Rapidly Progressive Dementia in CADASIL: A Case Report and Literature Review.

Cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL) is the most common hereditary cerebral small vessel disease with slow natural progression. Ischemic stroke and cognitive impairment are its most common clinical symptoms. Here, we report a rare 50-year-old woman who had rapid disease progression with c.457C>T, p.Argl53Cys heterozygous mutation in exon 4 of NOTCH3 and discuss the possible reasons. Furthermore, we summarized the clinical and neuroimaging characteristics of 14 CADASIL patients with Arg153Cys mutation in exon 4. The proband suffered acute ischemic stroke 5 times in 5 months, followed by rapidly progressive dementia (RPD) and inability to live independently, though she didn't have vascular risk factors and had been under standardized secondary prevention therapy since the first stroke. Magnetic resonance imaging showed extensive white matter hyperintensities, numerous ischemic infarcts and microbleeds, and severe brain atrophy. Her elder brother and other patients with Arg153Cys mutation in exon 4 all did not progress so quickly. Her multiple strokes may be associated with the poor self-regulation of vessels, which may promote the occurrence of RPD. Antiplatelet and anticoagulant drugs were difficult to prevent ischemic strokes. Severe imaging findings may indicate rapid progression of CADASIL. In addition, we found that headache was a very frequent symptom in CADASIL patients with Arg153Cys mutation in exon 4, accounting for 76.9%. CADASIL can also appear to have rapid progression, as illustrated by our proband, which is worthy of clinicians' attention and intervention timely. Headache may present in a relatively higher proportion of CADASIL patients with Arg153Cys mutation in exon 4.

USP8 Mutations Associated with Cushing’s Disease Alter Protein Structure Dynamics

The adenomas in Cushing’s disease frequently exhibit mutations in exon 14, within a binding motif for the regulatory protein 14-3-3 located between the catalytic domain (DUB), responsible for ubiquitin hydrolysis, and the WW-like domain that mediates autoinhibition, resulting in constantly active USP8. The exact molecular mechanism of deubiquitinase activity disruption in Cushing’s disease remains unclear. To address this, Sanger sequencing of USP8 was performed to identify mutations in corticotropinomas. These mutations were subjected to computational screening, followed by molecular dynamics simulations to assess the structural alterations that might change the biological activity of USP8. Eight different variants of the USP8 gene were identified both within and outside the “hotspot” region. Six of these had previously been reported in Cushing’s disease, while two were detected for the first time in our patients with CD. One of the two new variants, initially classified as benign during screening, was found in the neighboring SH3 binding motif at a distance of 20 amino acids. This variant demonstrated pathogenicity patterns similar to those of known pathogenic variants. All USP8 variants identified in our patients caused conformational changes in the USP8 protein in a similar manner. The identified mutations, despite differences in annotation results—including evolutionary conservation assessments, automated predictor data, and variations in localization within exon 14—exhibit similar patterns of protein conformational change. This suggests a pathogenic effect that contributes to the development of CD.

Mutational Landscape of KIT Proto-Oncogene Coding Sequence in 62 Canine Cutaneous and Subcutaneous Mast Cell Tumors

Canine mast cell tumors (MCTs) are common skin neoplasms with varying biological behaviors. The KIT proto-oncogene plays a key role in the development of these tumors, and internal tandem duplications on exon 11 are usually associated with more aggressive behavior, increased local recurrence, and decreased survival time. However, apart from exons 8–11 and 17, there is limited understanding of the overall KIT mutational landscape in canine MCTs. This work aims to analyze the entire KIT coding sequence (21 exons) in a cohort of 62 MCTs, which included 38 cutaneous and 24 subcutaneous tumors, and potentially identify new variants. In addition to confirming previously reported activating KIT mutations in exons 8, 9, and 11, we identified new variants in exons 2, 3, 5, 16, and the 3′ untranslated region (UTR). Notably, these last variants include an amino acid change (Asp/His) in exon 16. Additionally, we confirmed a differential prevalence of KIT variants in cutaneous and subcutaneous MCTs. These findings enhance our understanding of the KIT proto-oncogene coding sequence and provide valuable information for future confirmatory studies.

A novel SBF1 missense mutation causes autosomal dominant Charcot–Marie–Tooth disease type 4B3

IntroductionWe present a case of autosomal dominant Charcot–Marie–Tooth disease type 4B3 (CMT4B3) in a family caused by a novel SBF1 missense mutation.MethodsTwo patients, a mother and daughter, were recruited from our hospital. Both exhibited early-onset symptoms, including distal muscle atrophy of the limbs, without cranial nerve involvement. Electromyography was performed to assess nerve amplitudes and conduction velocities. Whole-exome sequencing (WES) and Sanger sequencing were performed to identify genetic mutations.ResultsElectromyography revealed a significant decline in nerve amplitudes, while the nerve conduction velocities (NCVs) remained normal in the extremities. Sequencing identified a novel missense mutation (c.1398C > A, p.H466Q) in exon 13 of the SET binding factor 1 (SBF1) gene in both patients, indicating an autosomal dominant inheritance pattern.DiscussionPathogenicity and protein predictions suggest that the myotubularin-related protein 5 (MTMR5), encoded by the mutated SBF1, may possess an altered structure, resulting in disease. These findings will help expand the phenotypic and genetic spectrum of CMT4B3.

Analysis of a family with hypouricemia due to type Ⅰ xanthinuria

This article reports a patient presenting with"extremely low uric acid levels in blood and urine"clinically along with reviewing relevant literature to consider a diagnosis of xanthinuria. Peripheral blood samples of the patient and her family were further collected for xanthine dehydrogenase(XDH) gene sequencing, showing that the patient had compound heterozygous mutations in exon 19:c.1995_2006del12(p.His666_Gly669del) and exon 10:c.871G>T(p.Glu291*), however no mutations were found in the gene encoding MOCOS on chromosome 18, confirming the diagnosis of hereditary xanthinuria type Ⅰ.The patient's father, son and daughter carried heterozygous mutations in exon 19:c.1995_2006del12(p.His666_Gly669del), and the mother carried heterozygous mutations in exon 10:c.871G>T(p.Glu291*).Mutations in the XDH gene cause a lack of xanthine oxidoreductase function, which hinders the production of uric acid, leading to very low or undetectable levels in blood and urine. Patients present clinically with hematuria, renal colic, urolithiasis, and even acute renal failure. Through the diagnosis and treatment of this patient and literature review, the article aims to deepen the understanding of purine metabolism and uric acid production process, and improve the clinicians' diagnosis and treatment ability of hypouricemia.

Analysis of BRCA1 Exon 2 Mutations in FFPE Tissues of Bangladeshi Bangali Females with Breast Cancer

Background: Among all breast cancers, 5 to 10% are hereditary and related to mutation of BRCA1 and BRCA2 gene. To date, extensive researches have been carried out for detection of mutations in the exon2 of BRCA1 gene. Genetic study on these two genes from formalin-fixed paraffin-embedded tissue in Bangladeshi population has not been reported so far. Objective: To identify mutation in exon2 of BRCA1 gene from formalin-fixed paraffin-embedded tissue of adult Bangladeshi Bangali female patients with breast cancer. Methods: The research was cross-sectional descriptive type of study. The study was conducted on Adult Bangladeshi Bangali female patients with breast cancer aged more than 18 years. A questionnaire was developed to record information on socio-demographic and reproductive characteristics. The data was analyzed by SPSS (Statistical package for the social sciences) version 23.0. The whole sequence of exon2 of chromosome 17 was amplified by short range PCR using the Gotaq master mix and primer sequence. The amplicons were confirmed by 2% agarose gel electrophoresis and sequencing was done by Sanger sequencer. Data analysis was performed using Chromas® software version 2.33 and Mega 7 software. The quary sequences were compared with the NCBI database. Results: Average age of the breast cancer patients was 45.46 (± 11.52) and the mean age at menarche was 12.46 (± 0.646). In this study, new mutations along with known mutations were found in exon2 of BRCA1 gene of eight adult female breast cancer patients. The mutations are insertions (C>A, C>T, T>A, G>T, G>C, T>C, A>T, C>G), deletion (C_) and single nucleotide substitution (T-C, G-A, T-G, C-G, T-A, G-C, C-A) in types. Conclusions: The aim of the research was to identify mutation in exon2 of BRCA1 gene. Eight patients have mutation in their DNA sequence revealed from formalin-fixed paraffin-embedded tissue. Further studies are required to evaluate whether these mutation contribute to breast cancer or not. Identification of known mutation along with new mutation with this small sample size emphasizes the importance of exploration the genetic makeup of Bangladeshi population to develop a database for proper screening and genetic counseling of the disease.

Lung cancer drug development: From traditional chemotherapy to targeted and immunotherapies

Abstract. Lung cancer remains one of the deadliest types of cancer in the world, with the number of deaths due to lung cancer increasing dramatically year after year. In the same period, the approach to the treatment of lung cancer has changed dramatically over the past few decades, from systemic chemotherapeutic drugs-platinum-based drugs in the beginning to a wide range of newer therapeutic approaches such as targeted therapies and immunotherapies at the present time. This article reviews the development of drug therapy for lung cancer. By comparing the differences in mechanism of action and clinical side effects between traditional chemotherapeutic drugs and targeted drugs, we aim to highlight the importance of the development of new targeted drugs and new therapeutic means. In addition, the article also analyses the development and achievements of novel drugs targeting NSCLC with MET-14 exon mutations, and looks into the possible future therapeutic strategies, but there are some research gaps in combination therapy. Through an analysis of the development of lung cancer drugs, this paper aims to provide a more comprehensive perspective to provide guidance for the development and treatment of lung cancer drugs in the future, but there are still a lot of different efficacies brought about by the combination of drugs that have not been resolved, and future research can focus on the direction of the combination of therapies.

A new immunomagnetic microfluidic device for characterizing EGFR mutations in circulating tumor cells from patients withnon-small cell lung cancer

Incorporating precision oncology into cancer management has begun to improve clinical outcomes. Accurate sampling techniques that detect molecular aberrations are crucial for effective implementation. Circulating tumor cells (CTCs), derived from primary or metastatic sites and present in the blood, are proposed as useful diagnostic tools, though their use has been limited due to their rarity, especially in early-stage cancers. This study presents a novel immunomagnetic microfluidic device that efficiently isolates CTCs for analyzing epidermal growth factor receptor (EGFR) mutations in patients with non-small cell lung cancer (NSCLC). The device was designed and laser-cut from polymethylmethacrylate. Validation experiments involved spiking PC-9 cells (an established lung cancer cell line containing GLU 746-ALA 750 deletion mutations in exon 19 of the EGFR gene) into media and isolating these cells. Exons 18 – 21 of EGFR were amplified using a polymerase chain reaction to demonstrate the device’s rapid mutation detection capability. Next-generation sequencing was used to characterize these exons in a cohort of 38 NSCLC patients, successfully isolating CTCs from all. Among these patients, 30 (79%) had EGFR mutations, with exon 19 showing the highest mutation rate (87%) and exon 21 the highest point mutation rate (23%). Our device captured CTCs effectively in <1 h, enabling mutation detection. Further studies are needed to assess the prognostic significance of these mutations, but this technology has potential applications in various solid tumors.

NCMP-01. EARLY RECURRENCE OF GLIOBLASTOMA MULTIFORM INVOLVING MULTIPLE BRAIN REGIONS ASSOCIATED WITH MUTATIONS IN EXON 4 OF ISOCITRATE DEHYDROGENASE 1 COMPLICATED BY GADOLINIUM-INDUCED ACUTE KIDNEY FAILURE: A CASE REPORT

Abstract BACKGROUND Glioblastoma multiform (GBM) remains one of the most aggressive primary brain tumors with high recurrence rates. Case presentation We present a case of a 43-year-old female, Yira, diagnosed with early recurrence of GBM involving multiple brain regions, accompanied by mutations in exon 4 of isocitrate dehydrogenase 1 (IDH1), a marker associated with poor prognosis. Additionally, this case was complicated by gadolinium-induced acute kidney failure on postoperative day 3. The patient initially presented with progressive neurological symptoms, including headache and cognitive decline. Multiplanar multiecho MRI of the brain was performed with IV contrast in orthogonal plane and revealed a moderate sized multilobulated heterogeneously and predominantly peripherally enhancing mass lesion with irregular margins and central non enhancing areas of necrosis arising from the right corona radiata, insular cortex, external capsule, lentiform nucleus, temporal, and frontal white matter with blood products. Perilesional edema was seen. Effacement of sulcal spaces. Right sylvian fissure, compression over right ventricle and midline shift of 8mm toward left side. There was dilatation of the left lateral ventricle with periventricular cerebrospinal fluid ooze. Uncal herniation was right side. On MR spectroscopy large choline, creatinine levels were seen in the lesion with reduced N-acetyl aspartate levels as compared to the normal parenchyma. Findings were suggestive of neoplastic lesion mostly high-grade glioma. Polymerase chain reaction sequencing confirmed missense mutation R132H in exon 4 of isocitrate dehydrogenase 1 gene. Despite maximal safe surgical resection, the patient developed recurrent GBM within three months, with gadolinium-based contrast agent exposure leading to acute kidney injury. CONCLUSION The combination of aggressive tumor behavior and complications from contrast agent administration highlights the challenges in managing recurrent GBM. This case underscores the importance of vigilant monitoring for complications in GBM patients, particularly those with IDH1 mutations, and the need for further research into novel therapeutic strategies.

Identification and in vivo functional analysis of a novel missense mutation in GATA3 causing hypoparathyroidism, sensorineural deafness and renal dysplasia syndrome in a Chinese family.

Hypoparathyroidism, sensorineural deafness, and renal dysplasia (HDR) syndrome is a rare autosomal dominant genetic disease associated with mutations in the GATA3 gene, which encodes GATA3 that plays essential roles in vertebrate development. This study aimed to identify and report the pathogenic mutation in GATA3 in a Chinese family diagnosed with HDR syndrome and determine its functional impacts in vivo. The clinical features of a 25-year-old male patient with HDR syndrome and his parents were collected. GATA3 gene exome sequencing and Sanger sequencing were performed on the proband and his family, respectively. Functional analyses of GATA3 were performed using bioinformatics tools and zebrafish assays to determine pathogenicity and phenotype spectrum. A novel, heterozygous, missense mutation in exon 4 of the GATA3 gene, c.863 G > A, p.Cys288Tyr, in the proband and his mother who presented the complete HDR triad, was predicted to be deleterious by in silico tools. 3D structure modeling showed that the variant caused significant structural changes. In vivo studies using a zebrafish animal model revealed the deleterious impact of the variant on the gill buds, otoliths, and pronephros. We identified a novel missense mutation, GATA3 p.Cys288Tyr, within a family with HDR syndrome and delineated it as a loss-of-function variant in vivo. This expands the spectrum of GATA3 mutations associated with HDR syndrome in the Chinese population and mimics HDR-related changes in vivo.

A novel SERPINC1 c.119G>A (p.Cys40Tyr) mutation with variable clinical expression in an Indian family.

Hereditary antithrombin (AT) deficiency due to mutations in SERPINC1 is known to be the most severe form of thrombophilia. We report three members in a family with hereditary AT deficiency with a novel mutation in exon 2 of SERPINC1, that is c.119 G>A (p.Cys40Tyr). Two brothers presented with acute pulmonary thromboembolism (PTE) at 18 and 21 years of age, whereas their 58-year-old father did not have any thrombotic episode till date. The in-silico prediction of the variant was found to be highly damaging by PolyPhen-2, SIFT and MutationTaster. Clinical exome sequencing did not show any strong coinherited thrombophilia genes, except SERPINE1 -844 G>A variant in homozygous state in the two affected brothers as compared to the father who was heterozygous for this variant. The additive effect of SERPINE1 variant in the clinical expression in two siblings cannot be ruled out, in the absence of any other known environmental triggering factors.

DAAO Mutant Sites among Different Mice Strains and Their Effects on Enzyme Activity.

Previous studies reported that D-amino acid oxidase (DAAO) activity was closely associated with neuropathic pain, cognitive characteristics of schizophrenia and so on. To determine DAAO mutant sites in different strains of mice and their effects on enzyme activity, we successfully constructed a prokaryotic expression system for heterologous expression of DAAO in vitro. There were total five nucleotide mutations distributed in exons 2, 8, 9, 10 of C57 mice. Three mutations located on exons 8 and 9 were synonymous mutations and had no variation on the encoded amino acid. The remaining two mutations in exons 2 (V64A) and 10 (R295H) were non-synonymous mutations, which might affect enzymatic activity and protein structure of mDAAO. Based on the determination of the kinetic constants and IC50 of mDAAO mutants in vitro, the differences in amino acid levels at these two sites (V64A, R295H) increased the affinity of C57 DAAO with substrate and enhanced its catalytic efficiency. Besides, the IC50 value of C57 DAAO was less than that of Balb/c and other DAAO mutants (SUN: reducted by about 11.9%; CBIO: reducted by about 26.5%), which meant that the affinity of C57 DAAO with CBIO was higher.

The p.D417N variant of TUBB4A as a possible cause of hereditary spastic paraplegia: a case report

BackgroundTubulins are dimeric proteins expressed in all eukaryotic cells, serving as the fundamental building blocks of microtubule filaments. The TUBB4A gene encodes the protein β-tubulin. Mutations of TUBB4A have been associated with two neurodegenerative diseases with very different clinical characteristics: dystonia type 4 (DYT4) and Hypomyelination with Atrophy of the Basal Ganglia and Cerebellum (H-ABC). Several cases of patients with H-ABC or unclassified leukoencephalopathy show spastic ataxia with or without leukodystrophy in association with mutations in exon 5 of TUBB4A gene.Case presentationWe report the case of a 65-year-old woman who has been complaining of progressive difficulty in walking since childhood. The neurological examination revealed, among all, a spastic gait, difficulties in standing on the toes and heels, brisk deep tendon reflexes at upper limbs, clonic patellar reflexes, brisk ankle reflexes, as well as bilateral Babinski and Hoffmann sign, brisk jaw jerk and severe lower limb spasticity. NGS studies for inherited cases of spastic paraplegia revealed a novel variant of unknown significance (VUS) (c.1249A > G, p.D417N), in TUBB4A. To the best of our knowledge, this variant has not been reported in the literature or any databases in association with these diseases.ConclusionWe report the case of a patient carrying a variant of uncertain significance (VUS) of the TUBB4A gene, showing a progressive, spastic paraparesis, supporting the extension of the phenotypic spectrum up to include spastic paraplegia.

Genomic insights into the specialisation and selection of the Jinding duck

The domestication of ducks represents a pivotal evolutionary shift from the unguided propagation of wild species to deliberate human-mediated selection, culminating in distinct behavioural, morphological, and physiological traits that differentiate domesticated ducks from their wild counterparts. This transition has yielded breeds with traits fine-tuned to specific economic roles, such as egg production, meat yield, or dual-purpose functionality. Duck domestication plays a significant role in poultry production globally, meeting the growing demand for eggs and meat in various regions. Here, we focus on the Jinding Duck (JDD), a breed renowned for its prolific egg-laying traits. Employing whole-genome resequencing data from 325 individuals across five Chinese indigenous duck breeds, we aimed to dissect the unique population structure and assess the genetic diversity within the JDD cohort. The findings reveal the distinct genetic heritage of JDD, diverged from other domesticated breeds, and show a relative paucity of genetic diversity. A salient discovery was a 200 kb genomic interval containing three genes (NCF2, SMG7, and ARPC5) with almost exclusive haplotypes, which were inherited from Anas platyrhynchos or Anas zonorhyncha, impacting the morphological attributes of JDD. The study highlights a c.28G>A non−synonymous mutation in the first exon of the LAMC1 gene, which is potentially influencing feather morphology in JDD. Our findings suggest that unique blue eggshell colouration in JDD is likely attributable to variations within the promoter element of the ABCG2 gene, distinguishing it from other breeds. Moreover, the MAP7 and FHL1 genes emerge as significant factors in the laying performance of JDD. These genetic insights are not only crucial for improving the JDD breed but also provide valuable information that could be applied to duck breeding programmes worldwide, helping enhance productivity and meet international demands for high-efficiency poultry breeds.

Molecular Profile of BCR-ABL1 Negative Myeloproliferative Neoplasm in a Moroccan Population.

Myeloproliferative neoplasms (MPN) are associated with clonal hematopoiesis, genomic instability, hemostasis dysregulation, and immune response. Classic BCR-ABL1 negative myeloproliferative neoplasms (BCR-ABL1 negative MPN), including polycythemia vera (PV), essential thrombocythemia (ET), and primary myelofibrosis (PMF), are frequently associated with somatic abnormalities in JAK2, CALR, and MPL. Mutant clones induce an inflammatory immune response leading to immuno-thrombosis. The aim of this research was to investigate JAK2/STAT5 mutations in 49 Moroccan patients and to study their correlation with demographic and hematological parameters. Genomic DNA was isolated from peripheral blood samples, and JAK2V617F, JAK2 exon 12, CALR exon 9, and MPL exon 10 mutations were tested by qPCR, AS-PCR, and Sanger sequencing. Statistical analysis was performed using SPSS version 22. Our results showed that 33 (67.35%) of our cohort was mutated for one of the investigated genes. JAK2V617F was detected in 25 cases, as well as two previously described CALR mutations: c.1092_1143del52 and c.1154_1155insTTGTC. Mutations in JAK2 exon 12 (c.1622G>A/C and c.1641+6T>C) were observed and in PV and ET. MPL substitution c.1544G>T was detected in two cases. Mutations in JAK2, CALR, and MPL were mutually exclusive and benign single nucleotide variations/polymorphisms (SNV/SNP) have been identified in CALR and JAK2. The JAK2-PV subgroup was associated with higher red mass cell, hemoglobin, and hematocrit levels. Higher platelet counts correlated with the JAK2-ET subgroup, while higher leukocyte and neutrophil counts were associated with the JAK2-PMF subgroup. Complete blood count revealed hyperleukocytosis accompanied by hyperplateletosis and a lower level of hemoglobin in CALR-ET mutated patients, compared to JAK2-ET mutated or triple negative-ET cases. In conclusion, our study provides valuable insights into the prevalence and characteristics of JAK2, CALR, and MPL mutations in BCR-ABL1 negative MPNs in the Moroccan population, highlighting the importance of genetic characterization to optimize the clinical management of these diseases.

Effects of mutation in myostatin exon 3 on growth and carcass characteristics of Savak Akkaraman sheep

The myostatin gene (MSTN) is one of the main regulatory genes involved in muscle growth and development in many animals. This study aims to determine the variations in the exon 3 region associated with MSTN expression in Savak Akkaraman sheep and analyze their relationship with growth characteristics. Molecular analyses (RLFP, Real-Time PCR and DNA sequencing) were performed using DNA and RNA isolations from 70 different sheep blood and meat samples. As a result of the study, exon 3 region analysis was performed using NgomIV, AluI, DraI, HaeIII and MspI restriction endonucleases to determine the genetic diversity in Savak Akkaraman sheep breed. Two genotypes (MM and Mm) were detected for MSTN/MspI and a monomorphic mm genotype indicating the conserved exon 3 region for MSTN/HaeIII. Additionally, four new Single Nucleotide Polymorphisms (SNPs) were detected at different positions (164th, 204th, 347th, and 441th) in the working exon region. Two of these SNPs (204th T→G: isoleucine→ methionine and 347th C→A-G: proline→glutamine-arginine) caused amino acid changes. In expression studies, it was determined that MSTN gene expression was higher in female sheep than in male sheep (p<0.05). The MM genotype (MSTN/MspI) was significantly associated with increased weight gain and MSTN expression, while TT genotype at SNP 204 and GG genotype at SNP 347 was found to be associated with meat yield and growth traits (p<0.05). As a result, the genetic differences and functional effects of the new genotypes determined in this study may shed light on breeding strategies aiming to increase meat production.

Generation of CRISPR/Cas9 modified human iPSC line with correction of heterozygous mutation in exon 6 of the CaSR gene.

The calcium-sensing receptor (CaSR) gene encodes a cell membrane G protein-coupled receptor (GPCR) which has a key role in maintaining the extracellular Ca2+ homeostasis. We aimed at correcting the compound heterozygous mutation in the 6th [c.1656delA, p.I554SfsX73] and 7th [c.2217T > A, p.C739X] exons of the CASR gene which the original patient-derived iPSC line had. The mutation is associated with neonatal severe primary hyperparathyroidism of the patient. We generated and characterized a CRISP/Cas9-edited hiPSC line with the restored sequence in the sixth exon of the CASR gene, bearing only heterozygous mutation in the 7th exon. The results showed that the new genetically modified cell line has karyotype without abnormalities, typical hiPSCs morphology, characteristic expression of pluripotency markers, and ability to develop into three germ layers, and differentiates in chondrogenic, adipogenic, osteogenic directions. This new cell line will complement the existing pool of CaSR-mutated cell lines, a valuable resource for in-depth understanding of neonatal severe primary hyperparathyroidism. This will allow further exploration of the application of pharmacological drugs in the context of personalized medicine to correct Ca-homeostasis disorders.