Our results indicate that further elucidation of the causes of KTCN is needed.įor the in vitro splicing experiments, genomic DNA samples from an affected individual (KTCN-014-16) and from an unaffected individual not carrying mutations (KTCN-014-01) were amplified using primers specific for the regions of interest in DOCK9, IPO5, and STK24, as shown in the Table. However, because the mutation effect was observed in vitro, a definitive relationship between DOCK9 and KTCN phenotype could not be established. The exon skipping causes a premature stop codon, disrupting the functional domains of DOCK9 protein, which may alter the biological role of DOCK9 as a Cdc42 activator.īased on in vitro results, we demonstrated that c.2262A>C substitution in DOCK9, previously identified in KTCN-affected members of an Ecuadorian family, leads to a splicing aberration. In vitro splicing analysis revealed that only c.2262A>C in exon 20 of DOCK9 led to aberrant splicing, resulting in the changed ratio between two protein isoforms: a normal transcript and a transcript with exon skipping. After transfecting HeLa cells with each construct, total RNA samples were extracted, reverse transcribed, and amplified using specific primers. We generated expression constructs using patient DNA as a template corresponding to the wild-type and mutant alleles of DOCK9, IPO5, and STK24.
As the pathogenic consequences of these variants were not obvious, we performed in vitro splicing analyses to determine their functional significance. Previously, we identified heterozygous single base-pair substitutions in DOCK9, IPO5, and STK24, showing concurrent 100% segregation with the affected phenotype in an Ecuadorian family. A typical use of the NATO Phonetic Alphabet would be to spell out each letter in a word over the phone by saying, for example: "S as in Sierra" (or "S for Sierra"), "E as in Echo, Y as in Yankee, F as in Foxtrot, R as in Romeo, I as in India, E as in Echo, D as in Delta" to communicate the spelling of the name "Seyfried" correctly.Keratoconus (KTCN) is a degenerative disorder of the eye that is characterized by a conical shape and thinning of the cornea, resulting in impaired visual function.These are used to avoid misunderstanding due to difficult to spell words, different pronunciations or poor line communication. Spelling alphabets, such as the NATO Phonetic Alphabet, consists of a set of words used to stand for alphabetical letters in oral communication.The NATO Phonetic Alphabet is instead a spelling alphabet (also known as telephone alphabet, radio alphabet, word-spelling alphabet, or voice procedure alphabet). Phonetic alphabets are used to indicate, through symbols or codes, what a speech sound or letter sounds like. Contrary to what its name suggests, the NATO Phonetic Alphabet is not a phonetic alphabet.military and has also been adopted by the FAA (American Federal Aviation Administration), ANSI (American National Standards Institute), and ARRL (American Radio Relay League). Thus this alphabet can be reffered as the ICAO/ITU/NATO Phonetic Alphabet or International Phonetic Alphabet. The NATO (North Atlantic Treaty Organization) Phonetic Alphabet is currently officially denoted as the International Radiotelephony Spelling Alphabet (IRSA) or the ICAO (International Civil Aviation Organization) phonetic alphabet or ITU (International Telecommunication Union) phonetic alphabet.
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