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TitleTnpB structure reveals minimal functional core of Cas12 nuclease family.
Journal, issue, pagesNature, Vol. 616, Issue 7956, Page 384-389, Year 2023
Publish dateApr 5, 2023
AuthorsGiedrius Sasnauskas / Giedre Tamulaitiene / Gytis Druteika / Arturo Carabias / Arunas Silanskas / Darius Kazlauskas / Česlovas Venclovas / Guillermo Montoya / Tautvydas Karvelis / Virginijus Siksnys /
PubMed AbstractThe widespread TnpB proteins of IS200/IS605 transposon family have recently emerged as the smallest RNA-guided nucleases capable of targeted genome editing in eukaryotic cells. Bioinformatic analysis ...The widespread TnpB proteins of IS200/IS605 transposon family have recently emerged as the smallest RNA-guided nucleases capable of targeted genome editing in eukaryotic cells. Bioinformatic analysis identified TnpB proteins as the likely predecessors of Cas12 nucleases, which along with Cas9 are widely used for targeted genome manipulation. Whereas Cas12 family nucleases are well characterized both biochemically and structurally, the molecular mechanism of TnpB remains unknown. Here we present the cryogenic-electron microscopy structures of the Deinococcus radiodurans TnpB-reRNA (right-end transposon element-derived RNA) complex in DNA-bound and -free forms. The structures reveal the basic architecture of TnpB nuclease and the molecular mechanism for DNA target recognition and cleavage that is supported by biochemical experiments. Collectively, these results demonstrate that TnpB represents the minimal structural and functional core of the Cas12 protein family and provide a framework for developing TnpB-based genome editing tools.
External linksNature / PubMed:37020015
MethodsEM (single particle)
Resolution2.8 - 3.14 Å
Structure data

16016

8bf8

EMDB-16016, PDB-8bf8:
ISDra2 TnpB in complex with reRNA
Method: EM (single particle) / Resolution: 2.8 Å

28655

8ex9

EMDB-28655, PDB-8ex9:
ISDra2 TnpB in complex with reRNA and cognate DNA, conformation 2 (RuvC domain unresolved)
Method: EM (single particle) / Resolution: 2.96 Å

28656

8exa

EMDB-28656, PDB-8exa:
ISDra2 TnpB in complex with reRNA and cognate DNA, conformation 1 (RuvC domain resolved)
Method: EM (single particle) / Resolution: 3.14 Å

Source
  • Deinococcus radiodurans (radioresistant)
  • deinococcus radiodurans r1 (radioresistant)
  • synthetic construct (others)
KeywordsRNA BINDING PROTEIN / Transposon / TnpB / reRNA / RuvC domain / Cas12 / IS200/IS605

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