Por el memento seguimos tratando de optimizar las condiciones para transfectar el promotor de CHDL-1 en el vector pGL4 utilizando nuevas enzimas de restricción y tratando de obtener la mayor cantidad posible del vector cortado. Además volvimos amplificar los diferentes fragmentos de la región promotora que pronto intentaremos clonar nuevamente al vector. Mientras tanto he aprendido una nueva técnica de laboratorio llamada “electrophoretic mobility shift assay” (EMSA). Esta técnica es basada en la observación de que los complejos de ADN-proteína migran más lento que las moléculas de ADN libre cuando se corren en un gel de electroforesis. Esta técnica nos ayudara a identificar la interacción de la región promotora de CHDL-1 con TWIST 2 y otras proteínas de interés.
Como solo llevo un semestre trabajando con este nuevo proyecto titulado “Mechanisms of gene regulation by the bHLH transcription factor TWIST 2”, todavía no he obtenido suficiente data para presentar en un poster. Decidí por lo tanto presentar mi proyecto anterior con el cual originalmente comencé en Biominds. Aquí les presento el abstract del poster que presentaré el sábado, 21 de marzo en el “Annual Presentation Day”.
Prevalence of CYP2C19 Gene Polymorphisms in the Puerto Rican Newborn Population
P. Silen-Rivera1, J. Duconge2, P. Piovanetti1, L. M. Castro-Rosario1, J. Y. Renta1, P. J. Santiago Borrero3, John Guerra3 and C. L. Cadilla1
School of Medicine, University of Puerto Rico Medical Sciences Campus. School of Pharmacy2, University of Puerto Rico Medical Sciences Campus1 Depts. of Biochemistry1 and Pediatrics3
Polymorphisms in the cytochrome P450 2C19 gene (CYP2C19 *2, splicing defect 681G>A, and *3, stop codon 636G>A) significantly alter the metabolism of some prescription drugs. We determined the frequencies of these alleles and genotypes, single and double carriers, in a Puerto Rican cohort and tested for the Hardy-Weinberg equilibrium assumption. A total of 122 DNA samples from the Puerto Rican Newborn Screening Program were genotyped using both PCR-based restriction fragment length polymorphism (RFLP) method and the physiogenomic (PG)-array technology. The polymorphism frequency for CYP2C19*2 allele was 0.139. The prevalence of genotypes for this clinically relevant mutation was 24.6% single carrier (heterozygous) and 1.64% double-carrier. Genotypes met Hardy-Weinberg equilibrium (p>0.01). The findings were confirmed by PG analysis using Illumina BeadArrayTM technology. No carriers for CYP2C19*3 mutation were detected, suggesting a very low frequency for this allele in the Puerto Rican population. This variant is considered rare in non-Asian descendant population. Considering the observed prevalence of CYP2C19*2 polymorphisms in Puerto Ricans, it is necessary to make local healthcare providers aware of the CYP2C19-related pharmacogenetics in order to achieve better clinical outcomes by developing DNA-guided personalized algorithms. This project has received support from NIH grants R25GM61838 and P20RR011126, as well as from the PRNSP of the UPR School of Medicine and the Bio-Minds program.
