The persistence infection of low-risk type （type 6 or type 11） of human papillomavirus （HPV） is the main cause of genital warts. Given the high rate of recurrence after treatment, the use of a new molecular agent ...The persistence infection of low-risk type （type 6 or type 11） of human papillomavirus （HPV） is the main cause of genital warts. Given the high rate of recurrence after treatment, the use of a new molecular agent is certain to be of value. The aim of this study was to achieve targeted inactivation of viral E7 gene in keratinocytes using the reprogrammed clustered regularly interspaced short palindromic repeats （CRISPR）/CRISPR-associated （Cas） 9 system. To accomplish this, a universal CRISPR-Cas9 system for targeting both HPV6/11 E7 genes was constructed by using a dual guide RNA vector. After transfection of the vector into E7-transfromed keratinocytes, the expression level of E7 protein was measured using western-blot analysis and the sequence of the E7 gene was determined using Sanger sequencing. Cell proliferation was analyzed by CCK-8 assay, and cell apoptosis was evaluated by Hoechst 33258 staining, flow cytometry analysis and ELISA assay. The results indicated that both HPV6/11 E7 genes can be inactivated by the single CRISPR-Cas9 system. Furthermore, silencing of E7 led to inhibition of cell proliferation and induction of apoptosis in E7-transfromed keratinocytes but not in normal keratinocytes. Our data suggested that the reprogrammed CRISPR-Cas9 system has the potential for the development of an adjuvant therapy for genital warts.展开更多
Synthetic biology is an emerging field focusing on engineering genetic devices and biomolecular systems for a variety of applications from basic biology to biotechnology and medicine.Thanks to the tremendous advances ...Synthetic biology is an emerging field focusing on engineering genetic devices and biomolecular systems for a variety of applications from basic biology to biotechnology and medicine.Thanks to the tremendous advances in genomics and the chemical synthesis of DNA in the past decade,scientists are now able to engineer genetic devices and circuits for cancer research and intervention,which offer promising therapeutic strategies for cancer treatment.In this article,we provide a systemic review on recent development achieved by the synthetic biologists,oncologists and clinicians of one National“973”Plan.We expand the synthetic biology toolkits involving DNA,RNA and protein bio-parts to explore various issues in cancer research,such as elucidation of mechanisms and pathways,creation of new diagnostic tools and invention of novel therapeutic approaches.We claimed that the Chinese synthetic biologists are promoting the basic research productions of tumor synthetic biology into the clinic.展开更多
文摘The persistence infection of low-risk type （type 6 or type 11） of human papillomavirus （HPV） is the main cause of genital warts. Given the high rate of recurrence after treatment, the use of a new molecular agent is certain to be of value. The aim of this study was to achieve targeted inactivation of viral E7 gene in keratinocytes using the reprogrammed clustered regularly interspaced short palindromic repeats （CRISPR）/CRISPR-associated （Cas） 9 system. To accomplish this, a universal CRISPR-Cas9 system for targeting both HPV6/11 E7 genes was constructed by using a dual guide RNA vector. After transfection of the vector into E7-transfromed keratinocytes, the expression level of E7 protein was measured using western-blot analysis and the sequence of the E7 gene was determined using Sanger sequencing. Cell proliferation was analyzed by CCK-8 assay, and cell apoptosis was evaluated by Hoechst 33258 staining, flow cytometry analysis and ELISA assay. The results indicated that both HPV6/11 E7 genes can be inactivated by the single CRISPR-Cas9 system. Furthermore, silencing of E7 led to inhibition of cell proliferation and induction of apoptosis in E7-transfromed keratinocytes but not in normal keratinocytes. Our data suggested that the reprogrammed CRISPR-Cas9 system has the potential for the development of an adjuvant therapy for genital warts.
文摘Synthetic biology is an emerging field focusing on engineering genetic devices and biomolecular systems for a variety of applications from basic biology to biotechnology and medicine.Thanks to the tremendous advances in genomics and the chemical synthesis of DNA in the past decade,scientists are now able to engineer genetic devices and circuits for cancer research and intervention,which offer promising therapeutic strategies for cancer treatment.In this article,we provide a systemic review on recent development achieved by the synthetic biologists,oncologists and clinicians of one National“973”Plan.We expand the synthetic biology toolkits involving DNA,RNA and protein bio-parts to explore various issues in cancer research,such as elucidation of mechanisms and pathways,creation of new diagnostic tools and invention of novel therapeutic approaches.We claimed that the Chinese synthetic biologists are promoting the basic research productions of tumor synthetic biology into the clinic.
