Synthetic Modified RNA Transfection Protocol

Synthetic modified RNA (modRNA) was generated as described [29 (link)]. Briefly, RNA was synthesized with the MEGAscript T7 kit (Ambion, Austin, TX). A custom ribonucleoside blend was used, comprising 6 mM 5’ cap analog (New England Biolabs), 7.5 mM adenosine triphosphate and 1.5 mM guanosine triphosphate (USB, Cleveland, OH), 7.5 mM 5-methylcytidine triphosphate and 7.5 mM pseudo-uridine triphosphate (TriLink Biotechnologies, San Diego, CA). Transfections of modRNA and multiple siRNA targeted against Sirt1 and Satb2 (both from Santa Cruz) were carried out with RNAiMAX (Invitrogen), as per the manufacturer’s instructions.

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Sadegh C., Ebina W., Arvanites A.C., Davidow L.S., Rubin L.L, & Macklis J.D. (2021). Synthetic modified Fezf2 mRNA (modRNA) with concurrent small molecule SIRT1 inhibition enhances refinement of cortical subcerebral/corticospinal neuron identity from mouse embryonic stem cells. PLoS ONE, 16(9), e0254113.

Publication 2021

MEGAscript T7 kit 5-methylcytidine triphosphate pseudo-uridine triphosphate RNAiMAX

Adenosine triphosphate Austin Guanosine triphosphate Ribonucleoside Sirna Sirt1 Transfections Triphosphate Uridine triphosphate

Corresponding Organization : Harvard Stem Cell Institute

Other organizations : Massachusetts General Hospital, Boston Children's Hospital

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Variable analysis

independent variables

Transfections of modRNA
Multiple siRNA targeted against Sirt1 and Satb2

dependent variables

Not explicitly mentioned

control variables

Not explicitly mentioned

controls

No positive or negative controls were explicitly mentioned.

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