Multicomponent systems for site-specific genome modifications
AssigneeThe Regents of the University of California
InventorsKathleen Collins, Xiaozhu Zhang, Briana Van Treeck, Heather E. Upton, Sarah Palm, Jeremy McIntyre
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Abstract
The invention includes systems, compositions, and methods for the making of modular gene editing through reverse transcriptase related processes. Systems and methods that use modified nucleotides and peptides are specifically provided.
Claims14
A system for genome editing, comprising
- (i) at least one reverse transcriptase construct (RTC), said RTC comprising at least one reverse transcriptase module (RTC:
- RT-module) comprising an mRNA encoding a reverse transcriptase (RT), at least one reverse transcriptase construct 5′ module (RTC:
- 5′ module), and/or at least one reverse transcriptase construct 3′ module (RTC:
- 3′ module), and
- (ii) at least one gene insertion construct (GIC), said GIC comprising at least one RNA template suitable for reverse transcription by a polypeptide encoded by the at least one RTC, wherein the at least one gene insertion construct comprises at least one optional GIC:
- 5′ module, at least one GIC:
- payload module, and at least one GIC:
- 3′ module.
A method comprising de novo design of a 5′ module that recruits host machinery for second strand nicking and thus second strand synthesis, the method optionally providing efficiency of insertion gain by de novo design of the 5′ module to (a) include a predetermined length and position of rRNA, (b) have enhanced RZ folding, and/or (c) recruit host cell machinery.
Dependencies are inferred from the claim text.