Invention Application
- Patent Title: RAB ESCORT PROTEIN POTENCY ASSAY
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Application No.: US15806136Application Date: 2017-11-07
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Publication No.: US20180135097A1Publication Date: 2018-05-17
- Inventor: John Fraser WRIGHT , Marina SUMAROKA
- Applicant: Spark Therapeutics, Inc.
- Applicant Address: US PA Philadelphia
- Assignee: Spark Therapeutics, Inc.
- Current Assignee: Spark Therapeutics, Inc.
- Current Assignee Address: US PA Philadelphia
- Main IPC: C12Q1/48
- IPC: C12Q1/48

Abstract:
Methods for measuring REP-1 and REP-2 activity are provided. In certain embodiments, a method includes: (a) contacting cells that do not express endogenous functional REP-1 or REP-2 protein with an adeno-associated viral (AAV) vector comprising a CHM gene encoding a REP-1 protein or CHM like gene encoding a REP-2 protein under conditions allowing cell transduction; (b) incubating transduced cells under conditions allowing expression of the encoded REP-1 or REP-2 protein; (c) lysing the transduced cells to produce an extract comprising the encoded REP-1 or REP-2 protein and Rab small GTPase (Rabs); (d) incubating said extract with a Rab substrate for a period of time and under conditions allowing prenylation of the Rab thereby forming prenylated Rab; and (e) detecting and/or quantifying the prenylated Rab, wherein the amount of prenylated Rab reflects REP-1 or REP-2 activity thereby measuring REP-1 or REP-2 activity.
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