Chemical Dual End-Labeling of Large Ribozymes

Ahunbay, Esra; Steffen, Fabio D; Zelger-Paulus, Susann; Sigel, Roland K. O.

doi:10.1007/978-1-0716-2047-2_13

Cited by 2 publications

(2 citation statements)

References 43 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Third, many biochemical techniques, such as FRET, have limited options on where the probe can be placed and hence are mostly confined to the termini, thus restricting complete characterization of the molecule. 5 One way to overcome both the size limitations of NMR and site-specific incorporation of modified nucleotides is to construct RNA segments sequentially, each of which can be manipulated for selective incorporation of isotopes (termed segmental labeling) and/or modified rNTPs and analogues 6 . There are three methods currently in use for segmental and site-specific labeling.…”

Section: Introductionmentioning

confidence: 99%

Extending the toolbox for RNA biology with SegModTeX: A polymerase-driven method for site-specific and segmental labeling of RNA

Haslecker

Pham

Glänzer

et al. 2023

Preprint

View full text Add to dashboard Cite

RNA performs a wide range of functions regulated by its structure, dynamics, and often post-transcriptional modifications. While NMR is the leading method for understanding RNA structure and dynamics, it is currently limited by the inability to reduce spectral crowding by efficient segmental labeling. Furthermore, because of the challenging nature of RNA chemistry, the tools being developed to introduce site- specific modifications are increasingly complex and laborious. Here we use a previously designed Tgo DNA polymerase mutant to present SegModTeX — a versatile, one-pot, copy-and-paste approach to address these challenges. By precise, stepwise construction of a diverse set of RNA molecules, we demonstrate the technique to be superior to RNA polymerase driven and ligation methods owing to its substantially high yield, fidelity, and selectivity. We also show the technique to be useful for incorporating fluorescent- and a wide range of other probes, which significantly extends the toolbox of RNA biology in general.

show abstract

Section: Introductionmentioning

confidence: 99%

Extending the toolbox for RNA biology with SegModTeX: A polymerase-driven method for site-specific and segmental labeling of RNA

Haslecker

Pham

Glänzer

et al. 2023

Preprint

View full text Add to dashboard Cite

show abstract

“…Second, with current methods, we cannot easily construct functionally relevant chemical states of RNAs; for example, with existing enzymatic approaches, we cannot add at specific sites N 6 -methyl-adenosine (m 6 A), 5-methyl-cytosine (m 5 C), pseudouridine (Ψ), etc. Third, many biochemical techniques, such as FRET, have limited options on where the probe can be placed and hence are mostly confined to the termini, thus restricting complete characterization of the molecule 5 .…”

Section: Introductionmentioning

confidence: 99%

Extending the toolbox for RNA biology with SegModTeX: a polymerase-driven method for site-specific and segmental labeling of RNA

Haslecker,

Pham,

Glänzer

et al. 2023

Nat Commun

View full text Add to dashboard Cite

RNA performs a wide range of functions regulated by its structure, dynamics, and often post-transcriptional modifications. While NMR is the leading method for understanding RNA structure and dynamics, it is currently limited by the inability to reduce spectral crowding by efficient segmental labeling. Furthermore, because of the challenging nature of RNA chemistry, the tools being developed to introduce site-specific modifications are increasingly complex and laborious. Here we use a previously designed Tgo DNA polymerase mutant to present SegModTeX — a versatile, one-pot, copy-and-paste approach to address these challenges. By precise, stepwise construction of a diverse set of RNA molecules, we demonstrate the technique to be superior to RNA polymerase driven and ligation methods owing to its substantially high yield, fidelity, and selectivity. We also show the technique to be useful for incorporating some fluorescent- and a wide range of other probes, which significantly extends the toolbox of RNA biology in general.

show abstract

Chemical Dual End-Labeling of Large Ribozymes

Cited by 2 publications

References 43 publications

Extending the toolbox for RNA biology with SegModTeX: A polymerase-driven method for site-specific and segmental labeling of RNA

Extending the toolbox for RNA biology with SegModTeX: A polymerase-driven method for site-specific and segmental labeling of RNA

Extending the toolbox for RNA biology with SegModTeX: a polymerase-driven method for site-specific and segmental labeling of RNA

Contact Info

Product

Resources

About