Ahmed F. Kilani
University of California, Berkeley
Network
Latest external collaboration on country level. Dive into details by clicking on the dots.
Publication
Featured researches published by Ahmed F. Kilani.
Journal of Biological Chemistry | 2002
Tianhong Zhou; Joseph Kim; Ahmed F. Kilani; Kihoon Kim; Walter Dunn; Solomon Jo; Edward Nepomuceno; Fenyong Liu
External guide sequences (EGSs) are small RNA molecules that bind to a target mRNA, form a complex resembling the structure of a tRNA, and render the mRNA susceptible to hydrolysis by RNase P, a tRNA processing enzyme. An in vitro selection procedure was used to select EGSs that direct human RNase P to cleave the mRNA encoding thymidine kinase (TK) of herpes simplex virus 1. One of the selected EGSs, TK17, was at least 35 times more active in directing RNase P in cleaving TK mRNAin vitro than the EGS derived from a natural tRNA sequence. TK17, when in complex with the TK mRNA sequence, resembles a portion of tRNA structure and exhibits an enhanced binding affinity to the target mRNA. Moreover, a reduction of 95 and 50% in the TK expression was found in herpes simplex virus 1-infected cells that expressed the selected EGS and the EGS derived from the natural tRNA sequence, respectively. Our study provides direct evidence that EGS molecules isolated by the selection procedure are effective in tissue culture. These results also demonstrate the potential for using the selection procedure as a general approach for the generation of highly effective EGSs for gene-targeting application.
Journal of Biological Chemistry | 2003
Hua Zou; Jarone Lee; Sean Umamoto; Ahmed F. Kilani; Joseph Kim; Phong Trang; Tianhong Zhou; Fenyong Liu
By using an in vitro selection procedure, we have previously isolated RNase P ribozyme variants that efficiently cleave an mRNA sequence in vitro. In this study, a ribozyme variant was used to target the overlapping region of the mRNAs encoding human cytomegalovirus (HCMV) major transcription regulatory proteins IE1 and IE2. The variant is about 90 times more efficient in cleaving the IE1/IE2 mRNA sequence in vitro than the ribozyme derived from the wild type RNase P ribozyme. Our results provide the first direct evidence that a point mutation at nucleotide position 80 of RNase P catalytic RNA from Escherichia coli (U80→ C80) increases the rate of chemical cleavage, and another mutation at nucleotide position 188 (C188→ U188) enhances substrate binding of the ribozyme. Moreover, the variant is more effective in inhibiting viral IE1 and IE2 expression and growth in HCMV-infected cells than the wild type ribozyme. A reduction of about 99% in the expression level of IE1 and IE2 and a reduction of 10,000-fold in viral growth were observed in cells that expressed the variant. In contrast, a reduction of less than 10% in IE1/IE2 expression and viral growth was observed in cells that either did not express the ribozyme or produced a catalytically inactive ribozyme mutant. Thus, engineered RNase P ribozyme variants are highly effective in inhibiting HCMV gene expression and growth. These results also demonstrate the feasibility of engineering highly effective RNase P ribozymes for gene targeting applications, including anti-HCMV gene therapy.
RNA | 1999
Ahmed F. Kilani; Fenyong Liu
RNase P ribozyme cleaves an RNA helix that resembles the acceptor stem and T-stem structure of its natural ptRNA substrate. When covalently linked with a guide sequence, the ribozyme can function as a sequence-specific endonuclease and cleave any target RNA sequences that base pair with the guide sequence. Using a site-directed ultraviolet (UV) cross-linking approach, we have mapped the regions of the ribozyme that are in close proximity to a substrate that contains the mRNA sequence encoding thymidine kinase of human herpes simplex virus 1. Our data suggest that the cleavage site of the mRNA substrate is positioned at the same regions of the ribozyme that bind to the cleavage site of a ptRNA. The mRNA-binding domains include regions that interact with the acceptor stem and T-stem and in addition, regions that are unique and not in close contact with a ptRNA. Identification of the mRNA-binding site provides a foundation to study how RNase P ribozymes achieve their sequence specificity and facilitates the development of gene-targeting ribozymes.
Journal of Clinical Virology | 2002
Phong Trang; Ahmed F. Kilani; Jarone Lee; Amy W. Hsu; Kwa Liou; Joe Kim; Arash Nassi; Kihoon Kim; Fenyong Liu
Ribozymes are promising gene-targeting agents for regulation of gene expression. In our recent studies, RnaseP (M1GS) ribozymes were constructed to target the overlapping region (IE mRNA) of IE1 and IE2 mRNAs of human cytomegalovirus (HCMV) and the mRNA (TK mRNA) coding for thymidine kinase (TK) of herpes simplex virus 1 (HSV-1). Our results indicate that RNase P ribozymes efficiently cleaved the IE mRNA and TK mRNA sequences in vitro. Significant inhibitions (approximately 75-85%) of HCMV IE1/IE2 and HSV-1 TK expression were observed in cells that expressed these ribozymes while a reduction of less than 10% was found in cells that did not express the ribozymes or expressed a disabled one that contained mutations abolishing catalytic activity. Ribozyme variants, which cleaved a TK mRNA sequence in vitro more efficiently than the ribozyme derived from the wildtype RNase P sequence, were selected by an in vitro selection system. When the selected ribozymes were expressed in cultured cells, they were more effective in inhibiting viral IE1/IE2 and TK expression and viral growth than the wildtype ribozyme sequence. Our results provide the first direct evidence that RNase P ribozymes are highly effective in inhibiting HCMV gene expression and growth. Moreover, a selection system was developed for generating novel ribozyme variants that cleave a mRNA substrate efficiently in vitro. These results suggest that M1GS ribozyme-mediated inhibition of expression of viral genes can be used as a new approach for the studies of HCMV gene function and the treatment of HCMV infection.
RNA | 1997
Joseph J. Kim; Ahmed F. Kilani; Xiaoyan Zhan; Sidney Altman; Fenyong Liu
Journal of Biological Chemistry | 2000
Ahmed F. Kilani; Phong Trang; Stephen Jo; Amy W. Hsu; Joseph Kim; Edward Nepomuceno; Kwa Liou; Fenyong Liu
Journal of Molecular Biology | 2000
Phong Trang; Ahmed F. Kilani; Joseph Kim; Fenyong Liu
Nucleic Acids Research | 2001
Phong Trang; Jarone Lee; Ahmed F. Kilani; Joe Kim; Fenyong Liu
Journal of Molecular Biology | 2002
Phong Trang; Amy W. Hsu; Tianhong Zhou; Jarone Lee; Ahmed F. Kilani; Edward Nepomuceno; Fenyong Liu
Nucleic Acids Research | 2000
Amy W. Hsu; Ahmed F. Kilani; Kwa Liou; Jarone Lee; Fenyong Liu