title: Selective silencing of gene target expression by siRNA expression plasmids in human cervical cancer cells.
abstract: RNA interference is a natural mechanism to silence post-transcriptional gene expression in eukaryotic cells in which microRNAs act to cleave or halt the translation of target mRNAs at specific target sequences. Mature microRNAs, 19-25 nucleotides in length, mediate their effect at the mRNA level by inhibiting translation, or inducing cleavage of the mRNA target. This process is directed by the degree of complementary nucleotides between the microRNAs and the target mRNA; perfect complementary base pairing induces cleavage of mRNA, whereas several mismatches lead to translational arrest. Biological effects of microRNAs can be manipulated through the use of small interference RNAs (siRNAs) generated by chemical synthesis, or by cloning in molecular vectors. The cloning of a DNA insert in a molecular vector that will be transcribed into the corresponding siRNAs is an approach that has been developed using siRNA expression plasmids. These vectors contain DNA inserts designed with software to generate highly efficient siRNAs which will assemble into RNA-induced silencing complexes (RISC), and silence the target mRNA. In addition, the DNA inserts may be contained in cloning cassettes, and introduced in other molecular vectors. In this chapter we describe an attractive technology platform to silence cellular gene expression using specific siRNA expression plasmids, and evaluate its biological effect on target gene expression in human cervical cancer cells.
authors.affiliation: Direction of Chronic Infections and Cancer, Research Center in Infection Diseases, National Institute of Public Health, Av. Universidad No. 655, Colonia Santa María Ahuacatitlán, Cuernavaca, Morelos, 62100, Mexico, operalta@correo.insp.mx.
authors.forename: Oscar
authors.initials: O
authors.lastname: Peralta-Zaragoza
authors.forename: Faustino
authors.initials: F
authors.lastname: De-la-O-Gómez
authors.forename: Jessica
authors.initials: J
authors.lastname: Deas
authors.forename: Gloria
authors.initials: G
authors.lastname: Fernández-Tilapa
authors.forename: Geny Del Socorro
authors.initials: Gdel S
authors.lastname: Fierros-Zárate
authors.forename: Claudia
authors.initials: C
authors.lastname: Gómez-Cerón
authors.forename: Ana
authors.initials: A
authors.lastname: Burguete-García
authors.forename: Kirvis
authors.initials: K
authors.lastname: Torres-Poveda
authors.forename: Victor Hugo
authors.initials: VH
authors.lastname: Bermúdez-Morales
authors.forename: Mauricio
authors.initials: M
authors.lastname: Rodríguez-Dorantes
authors.forename: Carlos
authors.initials: C
authors.lastname: Pérez-Plasencia
authors.forename: Vicente
authors.initials: V
authors.lastname: Madrid-Marina
mesh_terms: D001483:Base Sequence; D015153:Blotting, Western; D045744:Cell Line, Tumor; D002498:Centrifugation; D003001:Cloning, Molecular; D004247:DNA / Q000378:metabolism; D004926:Escherichia coli / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D020868:Gene Silencing; D006801:Humans; D035683:MicroRNAs / Q000378:metabolism; D008969:Molecular Sequence Data; D051059:PTEN Phosphohydrolase / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics*; D034741:RNA, Small Interfering / Q000378:metabolism*; D015203:Reproducibility of Results; D015183:Restriction Mapping; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D014162:Transfection; D014170:Transformation, Genetic; D002583:Uterine Cervical Neoplasms / Q000235:genetics*
chemical_list: C524940:MIRN21 microRNA, human; D035683:MicroRNAs; D034741:RNA, Small Interfering; D004247:DNA; D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human