Application of Modified mRNA in Somatic Reprogramming to Pluripotency and Directed Conversion of Cell Fate

Abstract

Modified mRNA (modRNA)-based somatic reprogramming is an effective and safe approach that overcomes the genomic mutation risk caused by viral integrative methods. It has improved the disadvantages of conventional mRNA and has better stability and immunogenicity. The modRNA molecules encoding multiple pluripotent factors have been applied successfully in reprogramming somatic cells such as fibroblasts, mesenchymal stem cells, and amniotic fluid stem cells to generate pluripotent stem cells (iPSCs). Moreover, it also can be directly used in the terminal differentiation of stem cells and fibroblasts into functional therapeutic cells, which exhibit great promise in disease modeling, drug screening, cell transplantation therapy, and regenerative medicine. In this review, we summarized the reprogramming applications of modified mRNA in iPSC generation and therapeutic applications of functionally differentiated cells.

Human Induced Pluripotent Stem Cells: From Cell Origin, Genomic Stability, and Epigenetic Memory to Translational Medicine

“… 21 However, reprogramming using synthetic modified mRNAs requires serial transgene delivery, and blood cells seem to be difficult for reprogramming using the modified mRNA technology. 22 The co-delivery of microRNA-302s/367 with synthetic modified mRNAs into fibroblasts promoted the generation of iPSCs with a high efficiency, 23 which might be tested in blood cells.…”

Modified mRNA-Based Vaccines Against Coronavirus Disease 2019

“…Recently, substantial modifications of mRNA have been investigated for their extensive applications. A modRNA-based vaccine contains 5′ cap, 5′ UTR (untranslated region), 3′ UTR, poly-A tail, and open reading frame (ORF) 79 , 80 . modRNA can be fully transcribed in vitro from a linearized DNA plasmid template by the enzymatic reaction in a cell-free system 81 , 82 .…”

“…The plasmid acts as a DNA template that transcripts an mRNA using an RNA polymerase in in vitro transcription (IVT). Subsequently, DNase is added to degrade the plasmid template and a novel cap analog 83 – 85 . As natural 5′ cap, including m7G, competes with guanosine triphosphate (GTP), it risks uncapping or becoming inactive mRNA.…”

“…New cap analogs (P(1)-3′-O,7-dimethylguanosine-5′ P3-guanosine-5′ triphosphate) were designed, named antireverse cap analogs (ARCAs), that are not able to incorporate in the reverse orientation and skip degeneration by Dcp2 (mRNA-decaying enzymes) 83 . The incorporation of a cap analog can enhance translation efficiency and modRNA stability 65 , 83 , 87 – 89 . Substantial modification in mRNA was reported 90 , whereas only a small subset of modified nucleotides, such as 5-methylcytidine, pseudouridine, 5-methyluridine, N6-methyladenosine, N1-methyl-pseudouridine, and 5-methoxyuridine, were demonstrated to enhance translation efficiency and mRNA stability 65 , 77 , 88 , 91 – 93 .…”

Induced pluripotent stem cells: Generation methods and a new perspective in COVID-19 research

“…These cells are similar to ESCs in characteristics such as morphology, pluripotency, and marker expression. While ESCs are obtained from the blastocyst᾽s inner cell mass (ICM), iPSCs can be derived from a wide range of different cellular sources using different reprogramming methods ( Wang, 2021 ). In the earliest studies on iPSCs, the Yamanaka laboratory used different types of retroviruses to produce iPSCs, each carrying only one type of the transcription factor.…”