Method for expression of small RNA molecules within a cell

摘要

The invention provides methods and compositions for the expression of small RNA molecules within a cell using a lentiviral vector. The methods can be used to express doubles stranded RNA complexes. Small interfering RNA (siRNA) can be expressed using the methods of the invention within a cell, which are capable of down regulating the expression of a target gene through RNA interference. A variety of cells can be treated according to the methods of the invention including embryos, embryogenic stem cells, allowing for the generation of transgenic animals or animals constituted partly by the transduced cells that have a specific gene or a group of genes down regulated.

主权项

1. A retroviral construct for the expression of an siRNA within a cell, comprising:
a nucleic acid having the R and U5 sequences from a 5′ lentiviral long terminal repeat (LTR); a self-inactivating lentiviral 3′ LTR; and a first RNA Polymerase III promoter configured to operably linked to a first RNA coding region encoding a first siRNA; wherein the first RNA Polymerase III promoter is located between the 5′ LTR and the 3′ LTR.