The　host　range　of　human　immunodeficiency　virus　type　1　(HIV-1)　is　extremely　narrow,　which　has　hampered　the　establishment　of　non-human　primate　models　for　HIV-1　infection.　The　species-specific　innate　immune　factor　tripartite　motif　5　alpha　(TRIM5　alpha)　is　a　key　molecule　that　confers　potent　resistance　against　HIV-1　infection.　In　this　study,　we　targeted　the　TRIM5　alpha　gene　of　rhesus　macaques　(rhTRIM5　alpha)　using　the　transcription　activator-like　effector　nuclease　(TALEN)　to　study　the　effect　on　HIV-1　infection.　CD4(+)　T　cells　were　separated　from　the　peripheral　blood　of　rhesus　macaques　by　magnetic　cell　sorting,　and　the　positive　rate　was　greater　than　99%.　TALEN　plasmids　targeting　rhTRIM5　alpha　were　constructed　and　introduced　into　CD4(+)　T　cells　by　electroporation,　with　a　transfection　efficiency　of　approximately　25%.　The　genome　of　the　targeted　cells　was　extracted,　and　the　target　efficiency　was　analyzed　by　T7E1　enzyme　digestion.　After　sorting　the　positive　transductants,　the　TALENs　induced　rhTRIM5　alpha　mutations　at　a　rate　of　more　than　40%.　The　ability　of　the　HIV-1　virus　to　infect　the　targeted　cells　was　demonstrated　by　ELISA.　The　results　showed　that　targeting　rhTRIM5　alpha　enhanced　the　susceptibility　to　HIV-1　infection.　This　finding　will　pave　the　way　for　further　establishment　of　a　new　rhesus　macaque　model　for　HIV-1　studies.