Single lentiviral vector system for induced pluripotent (IPS) stem cells derivation

Abstract

The present invention is based on the discovery that a single lentiviral vector expressing multiple individual transcription factor proteins from a single multi-cistronic mRNA can reprogram a fibroblast cell to a stem cell-like cell. These reprogrammed induced pluripotent stem (iPS) cells are pluripotent. Additions of the Cre-LoxP sequences into the single lentiviral vector facilitate excision of the vector after reprogramming in achieved. Addition of a maker gene into the single lentiviral vector facilitates detection of the presence of the vector in an iPS. The invention provides compositions and methods of producing iPS cells using a single multi-cistronic lentiviral vector.

Description

CROSS REFERENCE TO RELATED APPLICATIONS[0001]This application is a continuation application of co-pending U.S. application Ser. No. 13 / 080,728 filed on Apr. 6, 2011, which is a continuation application of the International Application No. PCT / US2009 / 059660, filed Oct. 6, 2009, which designates the United States, which claims benefit under 35 U.S.C. §119(e) of the U.S. Provisional Application No. 61 / 103,091 filed on Oct. 6, 2008, the contents of which are incorporated herein by reference in their entirety.SEQUENCE LISTING[0002]The instant application contains a Sequence Listing which has been submitted electronically in ASCII format and is hereby incorporated by reference in its entirety. Said ASCII copy, created on Sep. 5, 2014, is named 701586-063733-C2_SL.txt and is 87,936 bytes in size.BACKGROUND OF INVENTION[0003]The capacity of embryonic stem cells (ESCs) to give rise to all types of somatic cells together with their ability to grow indefinitely in culture underscores their pot...

AI Technical Summary

Benefits of technology

The patent describes a method of making cells that can be used to create tissues or treat diseases. The method involves using a special tool to remove a part of the DNA that controls the cells' behavior. These cells are then able to grow and develop into different types of cells that can help with treating things like diabetes or cancer. The tool used to remove the DNA is a special tool that can cut and leave behind only the part that controls the cells. This makes the cells better and more likely to develop into specific types.

Problems solved by technology

However, many challenges exist, for example, ES cells are not genetically identical to the organism from which they are harvested, and thus rejection and immunogenicity are two concerns that potentially limit their future use in clinical transplantation.

However, the generation of patient-specific autologous ESCs is technically challenging and further complicated by ethical concerns, significantly limiting their potential for clinical transplantation.

This presence of multiple viral integrations across the genome makes their genetic elimination to produce safer iPS cells very difficult.

Moreover, many cells will receive only one, two or three factors, making it difficult to study the biochemistry of reprogramming on a homogeneous population of cells.

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