Greenlight Biosciences, Inc.

Massachusetts Institute of Technology

James Robbins Abshire

Klavs F Jensen

Armon R Sharei

Drew S Cunningham

Daniel MacEachran

Jacquin Clarence Niles

Mehak Gupta

William Jeremy Blake

Andrey J Zarur

Matthew Eduardo Moura

Naveen Sudharsan

Katherine J Williams

Ifeyinwa Iwuchukwu

Christian Matranga

Kelsey A Haugh

Himanshu Dhamankar

Nicholas Skizim

Rachit Jain

Karthikeyan Ramachandriya

The current subject matter includes methods, systems, articles, and techniques to deliver material to anucleate cells, such as red blood cells. Using a rapid deformation based microfluidic system, loading of red blood cells with macromolecules of different sizes has been shown. Although delivery to some mammalian cells, such as cancer cell lines and fibroblasts had been previously demonstrated using this technique, those designs were incompatible with RBCs that have dramatically different physical properties. Through the use of smaller constriction sizes, high speeds and different buffers successful delivery to red blood cells can be achieved. By enabling robust delivery to red blood cells in a simple, scalable manner, the current subject matter can be implemented in a diversity of applications that deliver material to study red blood cell diseases and/or use red blood cells as a therapeutic platform. Related apparatus, systems, techniques, and articles are also described.

Delivery of materials to anucleate cells

Provided herein, are compositions based on retroviruses (e.g., lentiviruses) comprising one or more nucleic acid molecules encoding retroviral Pol polyprotein components and a nucleic acid molecule comprising one or more transgene sequences flanked by long terminal repeat sequences, for delivery of the one or more transgenes to a target cell ex vivo or in vivo. The compositions are useful for delivering to a target cell (e.g., hematopoietic stem cells (HSCs), liver cells, ocular cells, muscle cells, epithelial cells, T cells, etc.) and/or stably expressing any transgene (e.g., beta-globin, Factor VIII, RP GTPase regulator (RPGR), dystrophin, cystic fibrosis transmembrane conductance regulator (CFTR), a chimeric antigen receptor, etc.) with a biological effect to treat and/or ameliorate the symptoms associated with any disorder related to gene expression (e.g., sickle cell disease, beta-thalassemia, haemophilia B, retinitis pigmentosa, Duchenne muscular dystrophy, cystic fibrosis, cancer, etc.).

Nucleic acid therapeutics for genetic disorders

Provided herein, in some aspects, are methods and compositions for cell-free production of ribonucleic acid.

Cell-free production of ribonucleic acid

Provided herein, in some embodiments, are methods and composition for the production of nucleoside triphosphates and ribonucleic acids.

Methods and compositions for nucleoside triphosphate and ribonucleic acid production

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Cambridge, MA, United States of America

James Robbins Abshire