Cold Spring Harbor Laboratory

Dana-farber-cancer Institute Inc.

Cold Spring Harbor Institute

Johannes Zuber

Scott William Lowe

Christopher Vakoc

James Elliott Bradner

Constantine S Mitsiades

Junwei Shi

Gregory J Hannon

Bruce W Stillman

Christof Fellmann

Anthony Mazurek

Katherine McJunkin

Scott W Lowe

Christopher R Vakoc

The invention provides compositions, methods, and kits for the treatment of acute myeloid leukemia in a subject.

Compositions and methods for treating leukemia

Provided is a single construct combining a sequence encoding an RNAi molecule, a sequence encoding a reporter, and a target sequence specific for the RNAi molecule. The construct can be used to determine the potency of the encoded RNAi molecule in a direct and unbiased way. These results can be used to inform the design of potent RNAi molecules of various types and can be extended to several other applications, including: (1) generation of tiled libraries comprising every possible RNAi molecule-encoding sequence for a given gene target; (2) large-scale parallel validation of RNAi molecules targeting many genes to generate validated RNAi molecule-encoding libraries; (3) experimental comparison of design algorithms and strategies; and (4) investigation of RNAi biology in target site mutagenesis assays by screening pools containing single nucleotide changes in target sites and/or in the RNAi molecule to identify the most relevant sequence characteristics of potent RNAi-target site predictions.

High throughput methods for functionally determining RNA interference efficiency

Provided is a mosaic mouse model for use in determining the potency of an shRNA in vivo for reducing survival of cancer cells of chemotherapy-resistant leukemia. The syngeneic mouse recipient is transplanted with tet-on competent leukemia cells carrying a bicistronic nucleic acid construct comprising a promoter operably linked to a fusion gene associated with chemotherapy-resistant leukemia, and a sequence encoding a reverse tet-transactivator protein, such that both coding sequences are co-expressed from the promoter. Also provided are methods of treating soft tissue cancers.

Identification of RNAI targets and use of RNAI for rational therapy of chemotherapy-resistant leukemia and other cancers

Growing community of inventors

Cold Spring Harbor, NY, United States of America

Johannes Zuber