Home  |  Archive  |  Online Submission  |  News & Events  |  Subscribe  |  APFA  |  Society  |  Links  |  Contact Us  |  中文版

Ahead of print
Authors' Accepted
Current Issue
Special Issues
Browse by Category

Manuscript Submission

Online Submission
Online Review
Instruction for Authors
Instruction for Reviewers
English Corner new!

About AJA

About AJA
Editorial Board
Contact Us

Resources & Services

Email alert

Download area

Copyright licence
EndNote style file
Manuscript word template
Guidance for AJA figures
    preparation (in English)

Guidance for AJA figures
    preparation (in Chinese)

Proof-reading for the

AJA Club (in English)
AJA Club (in Chinese)


Societies & Institutes
Databases & Libraries
Other links


Volume 18, Issue 3 (May 2016) 18, 475–479; 10.4103/1008-682X.157399

Reprogrammed CRISPR-Cas9 targeting the conserved regions of HPV6/11 E7 genes inhibits proliferation and induces apoptosis in E7-transformed keratinocytes

Yu-Chen Liu1, Zhi-Ming Cai2, Xue-Jun Zhang1

1 Institute of Dermatology, Department of Dermatology, The First Affiliated Hospital, Anhui Medical University, Hefei; Key Laboratory of Dermatology, Ministry of Education, State Key Laboratory of Dermatology Incubation Center, Department of Dermatology, Anhui Medical University, Hefei, China
    2 Key Laboratory of Medical Reprogramming Technology, Department of Urology, The Genitourinary Institution of Shenzhen University, Shenzhen Second People's Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen, China

Correspondence: Dr. XJ Zhang (ayzxj@vip.sina.com) or Dr. ZM Cai (caizhiming2000@163.com)



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 E 7 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 E 7 genes was constructed by using a dual guide RNA vector. After transfection of the vector into E 7-transfromed keratinocytes, the expression level of E 7 protein was measured using western-blot analysis and the sequence of the E 7 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 E 7 genes can be inactivated by the single CRISPR-Cas9 system. Furthermore, silencing of E 7 led to inhibition of cell proliferation and induction of apoptosis in E 7-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.

Keywords: Keywords: apoptosis; CRISPR-Cas9; HPV E 7; proliferation

Full Text | PDF | 中文摘要 |

Browse:  631
Copyright 1999-2017  Shanghai Materia Medica, Shanghai Jiao Tong University.  All rights reserved