The study explores the tuning of charge transport and spin dynamics in two-dimensional (2D) conjugated metal-organic frameworks (2D c-MOFs) by controlling interlayer stacking. By introducing bulky side groups on the conjugated ligands, the researchers achieve a transition from serrated to staggered stacking, reducing interlayer interactions and enhancing spin density. This results in a 30-fold increase in spin density and a significantly longer spin-lattice relaxation time (T1) of ~60 μs, outperforming reference 2D c-MOFs with compact stackings. The findings reveal that spinless polaron pairs or bipolarons play a crucial role in charge transport, and the strategy provides a bottom-up approach for enhancing spin dynamics in 2D c-MOFs, offering potential for spintronic applications.The study explores the tuning of charge transport and spin dynamics in two-dimensional (2D) conjugated metal-organic frameworks (2D c-MOFs) by controlling interlayer stacking. By introducing bulky side groups on the conjugated ligands, the researchers achieve a transition from serrated to staggered stacking, reducing interlayer interactions and enhancing spin density. This results in a 30-fold increase in spin density and a significantly longer spin-lattice relaxation time (T1) of ~60 μs, outperforming reference 2D c-MOFs with compact stackings. The findings reveal that spinless polaron pairs or bipolarons play a crucial role in charge transport, and the strategy provides a bottom-up approach for enhancing spin dynamics in 2D c-MOFs, offering potential for spintronic applications.