Rapid and reversible knockdown of endogenous proteins by peptide-directed lysosomal degradation.
Rapid and reversible methods for altering the levels of endogenous proteins are critically important for studying biological systems and developing therapeutics. Here we describe a membrane-permeant targeting peptide-based method that rapidly and reversibly knocks down endogenous proteins through chaperone-mediated autophagy in vitro and in vivo. We demonstrate the specificity, efficacy and generalizability of the method by showing efficient knockdown of various proteins, including death associated protein kinase 1 (160 kDa), scaffolding protein PSD-95 (95 kDa) and α-synuclein (18 kDa), with their respective targeting peptides in a dose-, time- and lysosomal activity-dependent manner in rat neuronal cultures. Moreover, we show that, when given systemically, the peptide system efficiently knocked down the targeted protein in the brains of intact rats. Our study provides a robust and convenient research tool for manipulating endogenous protein levels and may also lead to the development of protein knockdown-based therapeutics for treating human diseases.
1] Brain Research Centre and Department of Medicine, Vancouver Coastal Health Research Institute, University of British Columbia, Vancouver, British Columbia, Canada. [2].
Xuelai
X
Fan
1] Brain Research Centre and Department of Medicine, Vancouver Coastal Health Research Institute, University of British Columbia, Vancouver, British Columbia, Canada. [2].
Wu Yang
WY
Jin
Brain Research Centre and Department of Medicine, Vancouver Coastal Health Research Institute, University of British Columbia, Vancouver, British Columbia, Canada.
Jie
J
Lu
Institute of Pharmacology, Medicine College of Shandong University, Jinan, China.
Jin
J
Wang
1] Brain Research Centre and Department of Medicine, Vancouver Coastal Health Research Institute, University of British Columbia, Vancouver, British Columbia, Canada. [2] Translational Medicine Research Center, China Medical University Hospital, Taichung, Taiwan. [3] Graduate Institute of Immunology, China Medical University, Taichung, Taiwan.
Yu Tian
YT
Wang
D000818:Animals; D001343:Autophagy / Q000187:drug effects / Q000502:physiology; D035843:Biomedical Research / Q000379:methods*; D003188:Composite Resins / Q000378:metabolism*; D055785:Gene Knockdown Techniques; D008247:Lysosomes / Q000378:metabolism*; D010455:Peptides / Q000502:physiology*; D014176:Protein Biosynthesis / Q000502:physiology; D051381:Rats
C505390:BR 100; D003188:Composite Resins; D010455:Peptides