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G rna cap structure analog

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The G RNA Cap Structure Analog is a modified guanine nucleotide that can be used to cap the 5' end of in vitro-transcribed RNA. It serves as a structural analog of the natural mRNA cap, facilitating efficient translation of the RNA.

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Lab products found in correlation

Rna clean and concentrator kit, by Zymo Research (2 mentions) Poly a polymerase, by Thermo Fisher Scientific (2 mentions) Vaccinia virus enzymes, by New England Biolabs (2 mentions) T7 rna polymerase, by New England Biolabs (2 mentions)

Close spelling variants of this product

M7G(5')ppp(5')G RNA Cap Structure Analog (11 citations) 3′-O-Me-m7G(5′)ppp(5′)G RNA cap structure analog (8 citations) Cap structure analog (4 citations) G(5’)ppp(5’)A RNA cap structure analog (4 citations) G(5′)ppp(5′)G RNA cap structure analog (4 citations) 3′-0-Me-m7G(5′)ppp (5′)G RNA Cap Structure Analog (ARCA; (2 citations)

3 protocols using g rna cap structure analog

1

Characterization of RNA Cap Analogues

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The RNAs 1 to 4 were purchased from New England Biolab (Ipswich, MA, USA): RNA#1: G(5’) ppp(5’) A RNA Cap Structure Analog (S1406); RNA #2: m7 G(5’) ppp(5’) A RNA Cap Structure Analog (S1405); RNA #3: G(5’) ppp(5’) G RNA Cap Structure Analog (S1407); and RNA #4: m7 G(5’) ppp(5’) G RNA Cap Structure Analog (S1405). Other oligonucleotide (oligo RNAs) substrates were synthesized by TriLink Biotechnologies (Maravai LifeSciences, San Diego, CA, USA): Oligo #1: 5’ {Gppp.}A[mG] UUG UUA GUC UA 3’; Oligo #2: 5’ {Gppp.}AG UUG UUA GUC UA 3’; Oligo#3: 5’ {N7-MeGppp.}AG UUG UUA GUC UA 3’. Additional Oligo RNAs were obtained from IDT Technologies, Inc (Coralville, Iowa, USA): Oligo RNA 0011: 5’- UAC ACU CGA UCU GGA CUA AAG CUG CUC-3’; Oligo RNA 0012: 5’-ACG AGU CCU GGA CUG AAA CGG ACU UGU-3’; Oligo RNA 0020: 5’-ACA GAG A-3’; and Oligo RNA 0021: 5’-UAC AGA GAA-3.
Hsiao K., Zegzouti H, & Goueli S. (2022). High throughput bioluminescent assay to characterize and monitor the activity of SARS-CoV-2 methyltransferases. PLOS ONE, 17(11), e0274343.
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2

In Vitro Transcription and Purification of RNAs

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RNAs were made by in vitro transcription with T7 RNA polymerase (NEB). For luciferase RNAs, transcription was performed in the presence of 3′-O-Me-m7G(5′)ppp(5′)G RNA Cap Structure Analog (NEB), using linearized plasmid as the template, and poly-adenylated using polyA polymerase (Invitrogen). For gel shifts, annealed oligonucleotides were used as the template, and RNAs were radiolabeled by capping with vaccinia virus enzymes (NEB) and [α-32P]-GTP. For SHAPE reactions, PCR templates were made using primers to add a 3′ handle (5′- GAACCGGACCGAAGCCCGGGCTGAG-3′), and transcription was performed using gel-extracted PCR products. RNAs were purified by phenol-chloroform extraction and ethanol precipitation or using the RNA Clean and Concentrator Kit (Zymo).
Lee A.S., Kranzusch P.J, & Cate J.H. (2015). eIF3 targets cell proliferation mRNAs for translational activation or repression. Nature, 522(7554), 111-114.
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3

In Vitro Transcription and Purification of RNAs

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RNAs were made by in vitro transcription with T7 RNA polymerase (NEB). For luciferase RNAs, transcription was performed in the presence of 3′-O-Me-m7G(5′)ppp(5′)G RNA Cap Structure Analog (NEB), using linearized plasmid as the template, and poly-adenylated using polyA polymerase (Invitrogen). For gel shifts, annealed oligonucleotides were used as the template, and RNAs were radiolabeled by capping with vaccinia virus enzymes (NEB) and [α-32P]-GTP. For SHAPE reactions, PCR templates were made using primers to add a 3′ handle (5′- GAACCGGACCGAAGCCCGGGCTGAG-3′), and transcription was performed using gel-extracted PCR products. RNAs were purified by phenol-chloroform extraction and ethanol precipitation or using the RNA Clean and Concentrator Kit (Zymo).
Lee A.S., Kranzusch P.J, & Cate J.H. (2015). eIF3 targets cell proliferation mRNAs for translational activation or repression. Nature, 522(7554), 111-114.
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