This study explores the tunable charge transport and spin dynamics in two-dimensional conjugated metal-organic frameworks (2D c-MOFs) by controlling interlayer stacking. The research focuses on the effect of bulky side groups on the conjugated ligands, which leads to a transition from serrated stacking to staggered stacking, thereby weakening interlayer interactions. This results in a significant increase in spin density and spin-lattice relaxation time (T₁), up to ~60 μs, while decreasing electrical conductivity by six orders of magnitude. The staggered stacking also enhances spin transport, with spinless polaron pairs or bipolarons playing a critical role in charge transport. The study demonstrates that the spin density in Ni₃(HATI_iPr)₂ is 30 times higher than in Ni₃(HATI_H)₂, and its T₁ is significantly longer, indicating improved spin dynamics. The results highlight the importance of interlayer stacking in controlling spin and charge transport properties in 2D c-MOFs, offering a pathway for developing MOF-based spintronics. The findings suggest that precise control of stacking modes can enhance spin dynamics, making 2D c-MOFs promising candidates for spin qubits and spintronic applications. The study provides a bottom-up approach for designing 2D c-MOFs with tailored spin and charge transport properties.This study explores the tunable charge transport and spin dynamics in two-dimensional conjugated metal-organic frameworks (2D c-MOFs) by controlling interlayer stacking. The research focuses on the effect of bulky side groups on the conjugated ligands, which leads to a transition from serrated stacking to staggered stacking, thereby weakening interlayer interactions. This results in a significant increase in spin density and spin-lattice relaxation time (T₁), up to ~60 μs, while decreasing electrical conductivity by six orders of magnitude. The staggered stacking also enhances spin transport, with spinless polaron pairs or bipolarons playing a critical role in charge transport. The study demonstrates that the spin density in Ni₃(HATI_iPr)₂ is 30 times higher than in Ni₃(HATI_H)₂, and its T₁ is significantly longer, indicating improved spin dynamics. The results highlight the importance of interlayer stacking in controlling spin and charge transport properties in 2D c-MOFs, offering a pathway for developing MOF-based spintronics. The findings suggest that precise control of stacking modes can enhance spin dynamics, making 2D c-MOFs promising candidates for spin qubits and spintronic applications. The study provides a bottom-up approach for designing 2D c-MOFs with tailored spin and charge transport properties.