mutations in the mcph1  (CH Instruments)

Journal: Heliyon

Article Title: Functional analysis of a novel intronic variant of MCPH1 with autosomal recessive primary microcephaly

doi: 10.1016/j.heliyon.2024.e30285

Figure Lengend Snippet: Minigene assay for MCPH1 c.233+2T > G mutation and schematic diagram of the splicing pattern. A, The construction of a Minigene trapping vector. B, Results from gel electrophoresis of RT-PCR demonstrated the presence of bands for wild-type and mutant-type. The agarose gel electrophoresis results showed that the PCR product of pMini-CopGFP-WT exhibited a band at 362 bp, whereas pMini-CopGFP-MT displayed a band at 762 bp. C, Analysis of the minigene product through sequencing showed that the wild-type minigene formed a normal mRNA, but the c.233+2T > G substitution of MCPH1 caused a splicing abnormality, which eliminated the Intron 3 canonical splice site, leading to a 400bp insertion. D, The schematic diagram showed the splicing pattern of wild-type and mutant-type.

Article Snippet: The specific PCR primers for the MCPH1 mutation were designed by the Primer 5.0 software and synthesized by Sangon Biotech.

Techniques: Mini Gene Assay, Mutagenesis, Plasmid Preparation, Nucleic Acid Electrophoresis, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Sequencing

Journal: Heliyon

Article Title: Functional analysis of a novel intronic variant of MCPH1 with autosomal recessive primary microcephaly

doi: 10.1016/j.heliyon.2024.e30285

Figure Lengend Snippet: Pedigree and sequence analysis of the proband . A, The pedigree of the Family. The arrows indicate the probands; B,C,D,The mutation c.233+2T > G of MCPH1 was identified in the proband (Ⅱ: 3), his father (Ⅰ: 1), and mother (Ⅰ: 2). The proband has a genotype of GG, while both his father and mother have a genotype of TG.

Article Snippet: The specific PCR primers for the MCPH1 mutation were designed by the Primer 5.0 software and synthesized by Sangon Biotech.

Techniques: Sequencing, Mutagenesis

Journal: Heliyon

Article Title: Functional analysis of a novel intronic variant of MCPH1 with autosomal recessive primary microcephaly

doi: 10.1016/j.heliyon.2024.e30285

Figure Lengend Snippet: The MCPH1 c.233+2T > G mutation resulting in truncated proteins. A, Locations of c.233+2T > G in the MCPH1 gene and protein structure. Red arrows indicate the positions of the mutation. B, Predicted wild-type protein; C, Predicted mutant-type protein.

Article Snippet: The specific PCR primers for the MCPH1 mutation were designed by the Primer 5.0 software and synthesized by Sangon Biotech.

Techniques: Mutagenesis

Journal: Heliyon

Article Title: Functional analysis of a novel intronic variant of MCPH1 with autosomal recessive primary microcephaly

doi: 10.1016/j.heliyon.2024.e30285

Figure Lengend Snippet: Mutations in the MCPH1 gene with MCPH.

Article Snippet: The specific PCR primers for the MCPH1 mutation were designed by the Primer 5.0 software and synthesized by Sangon Biotech.

Techniques:

Journal: Open Medicine

Article Title: The analyses of human MCPH1 DNA repair machinery and genetic variations

doi: 10.1515/med-2024-0917

Figure Lengend Snippet: Annotation of Exon Expression of MCPH1 derived from the GTEx Analysis V8. The functional annotations of the rare variants p.Val10SerfsTer5 andp.Ser571Ter to the exons and tissue specific expression values.

Article Snippet: More recently, mutations in the MCPH1 were associated with congenital hearing impairment (CHI) [ , ], and previously in otitis media in the mice model [ ].

Techniques: Expressing, Derivative Assay, Functional Assay

Journal: Open Medicine

Article Title: The analyses of human MCPH1 DNA repair machinery and genetic variations

doi: 10.1515/med-2024-0917

Figure Lengend Snippet: Overview of MCPH1 disease-associated variants identified in different domains of the canonical transcript. (ENST00000344683.9)

Article Snippet: More recently, mutations in the MCPH1 were associated with congenital hearing impairment (CHI) [ , ], and previously in otitis media in the mice model [ ].

Techniques:

Journal: Open Medicine

Article Title: The analyses of human MCPH1 DNA repair machinery and genetic variations

doi: 10.1515/med-2024-0917

Figure Lengend Snippet: Protein altering variants in identified in MCPH1 in the gnomAD data, and their VEP and ClinVar annotations considered pathogenic, likely pathogenic or uncertain significance

Article Snippet: More recently, mutations in the MCPH1 were associated with congenital hearing impairment (CHI) [ , ], and previously in otitis media in the mice model [ ].

Techniques:

Journal: Open Medicine

Article Title: The analyses of human MCPH1 DNA repair machinery and genetic variations

doi: 10.1515/med-2024-0917

Figure Lengend Snippet: The two highlighted regions in purple show where MCPH1 functions. The MCPH1 forms parts of the MRN complex, in conjunction with SET, condensin II, SMC2, SMC4, NCAPD3 , NCAPH2 , and NCAPG2 . The NHEJ pathway is initiated in response to the formation of DNA DSBs induced by DNA-damaging agents, such as ionizing radiation. The DNA DSBs are recognized by the MRN complex, leading to ATM activation and ATM-dependent recruitment of several DNA damage checkpoints and repair proteins to DNA DSB sites.

Article Snippet: More recently, mutations in the MCPH1 were associated with congenital hearing impairment (CHI) [ , ], and previously in otitis media in the mice model [ ].

Techniques: Activation Assay