SARS-CoV-2, like different coronaviruses, builds a membrane-bound replication organelle (RO) to allow RNA replication1. The SARS-CoV-2 RO consists of double membrane vesicles (DMVs) tethered to the endoplasmic reticulum (ER) by skinny membrane connectors2, however the viral proteins and the host components concerned are at the moment unknown. Right here we establish the viral non-structural proteins (NSPs) that generate the SARS-CoV-2 RO. NSP3 and NSP4 generate the DMVs whereas NSP6, by way of oligomerization and an amphipathic helix, zippers ER membranes and establishes the connectors. The NSP6ΔSGF mutant, which arose independently within the α, β, γ, η, ι, and λ variants of SARS-CoV-2, behaves as a gain-of-function mutant with a better ER-zippering exercise. We recognized three principal roles for NSP6: to behave as a filter in RO-ER communication permitting lipid movement however limiting entry of ER luminal proteins to the DMVs, to place and set up DMV clusters, and to mediate contact with lipid droplets (LDs) through the LD-tethering advanced DFCP1-Rab18. NSP6 thus acts as an organizer of DMV clusters and may present a selective observe to refurbish them with LD-derived lipids. Importantly, each correctly fashioned NSP6 connectors and LDs are required for SARS-CoV-2 replication. Our findings, uncovering the organic exercise of NSP6 of SARS-CoV-2 and of different coronaviruses, have the potential to gasoline the seek for broad antiviral brokers.