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Phân tích hoạt động của ranh giới cromatin trong tế bào Drosophila
Tóm tắt
Ranh giới cromatin, còn được gọi là bộ cách, điều chỉnh hoạt động gen bằng cách tổ chức các miền cromatin hoạt động và ức chế và điều chỉnh các tương tác giữa tăng cường và promoter. Tuy nhiên, cơ chế hoạt động của các ranh giới vẫn chưa được hiểu rõ, một phần là do kiến thức hạn chế của chúng ta về các protein bộ cách và sự thiếu thốn các xét nghiệm chuẩn mà có thể so sánh các ranh giới khác nhau. Chúng tôi báo cáo ở đây sự phát triển của một xét nghiệm chặn tăng cường để nghiên cứu hoạt động của bộ cách trong các tế bào Drosophila nuôi cấy. Chúng tôi cho thấy rằng hoạt động của các bộ cách Drosophila khác nhau, bao gồm suHw, SF1, SF1b, Fab7 và Fab8, được hỗ trợ trong các tế bào này. Chúng tôi cũng chỉ ra rằng sự giảm mức thuốc của các yếu tố SuHw và dCTCF thông qua RNA chuỗi đôi (dsRNA) làm gián đoạn chức năng ngăn chặn tăng cường của suHw và Fab8, từ đó xác lập hiệu quả của việc sử dụng can thiệp RNA trong xét nghiệm dựa trên tế bào của chúng tôi để điều tra chức năng của bộ cách. Xét nghiệm ranh giới mới cung cấp một phương pháp định lượng và hiệu quả để phân tích cơ chế bộ cách và có thể được khai thác thêm trong các sàng lọc RNAi trên quy mô toàn bộ gen cho các thành phần bộ cách. Nó cung cấp một công cụ hữu ích bổ sung cho các phương pháp chuyển gen và di truyền để nghiên cứu loại yếu tố điều tiết quan trọng này.
Từ khóa
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