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US 10,947,534 B2Granted
RNA-guided DNA integration using Tn7-like transposons
AssigneeThe Trustees of Columbia University in the City of New York
InventorsSamuel Henry Sternberg, Sanne Eveline Klompe
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Abstract
In certain embodiments, the present systems and methods use Tn7-like transposons that encode CRISPR-Cas systems for programmable, RNA-guided DNA integration. For example, the CRISPR-Cas machinery directs the Tn7 transposon-associated proteins to integrate DNA downstream of a target site (e.g., a genomic target site) recognized by a guide RNA (gRNA).
Claims9
A system for RNA-guided DNA integration, the system comprising one or more vectors heterologous to Vibrio cholerae encoding:
- a) an engineered Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-CRISPR associated (Cas) (CRISPR-Cas) system, the engineered CRISPR-Cas system comprising:
- Cas5, Cas6, Cas7 and Cas8;
- and
- b) an engineered transposon 7-like (Tn7-like) transposon system, the engineered Tn7-like transposon system comprising:
- i) Transposon 7 protein A (TnsA) TnsA, ii) Transposon 7 protein B (Tns B), iii) Transposon 7 protein C (Tns C), and iv) transposition of integron protein Q (TniQ),
- wherein the engineered Tn7-like transposon system is derived from Vibrio cholerae Tn6677.
3 more elements
Dependencies are inferred from the claim text.