Table 1 DNA-barcoded AAV virus libraries.

Library name	AAV clones contained in the libraries	Amino acids to be investigated	Total no. of clones
	AAV strains	No. of mutants	No. of clones
AAV-Serotype-VBCLib
	AAV9	Control	100
	AAV2R585E	Control	10
	AAV1	Wild type	2
	AAV2	Wild type	2
	AAV3	Wild type	2
	AAV4*	Wild type	2
	AAV5	Wild type	2
	AAV6	Wild type	2
	AAV7	Wild type	2
	AAV8	Wild type	2
	AAVrh10	Wild type	2
	AAV1.9-1^†	Hybrid	2
	AAV1.9-3-5^†	Hybrid	2		132
AAV9-AA-VBCLib-1
	AAV9	Control	15
	AAV2R585E	Control	15
	AAV9 double alanine mutants	92	184 (2 each)	356–539	214
AAV9-AA-VBCLib-2
	AAV9	Control	15
	AAV2R585E	Control	15
	AAV9 double alanine mutants	38	76 (2 each)	540–615	106
AAV9-AA-VBCLib-3
	AAV9	Control	15
	AAV2R585E	Control	15
	AAV9 double alanine mutants	61	122 (2 each)	616–736	152
AAV9-AA-VBCLib-4
	AAV9	Control	18
	AAV2R585E	Control	18
	AAV9 double alanine mutants	119	238 (2 each)
	AAV2R585E hexapeptide mutants	11	22 (2 each)	See footnote^‡
	AAV2R585E.9-2	1	2		298
AAV2R585E-HP-VBCLib-1
	AAV9	Control	15
	AAV2R585E	Control	15
	AAV2R585E hexapeptide mutants	66	132 (2 each)	441–484^§	162
AAV2R585E-HP-VBCLib-2
	AAV9	Control	15
	AAV2R585E	Control	15
	AAV2	Wild type	2
	AAV2R585E hexapeptide mutants	59	177 (3 each)	571–604^§	209

^*We lost AAV4 viral particles significantly from the library during the viral stock purification procedure. Accordingly, Illumina sequence read numbers for AAV4 in the viral stock were several logs lower than others. Therefore, AAV4 data were not evaluated or included in the study.
^†AAV1.9-1 and AAV1.9-3-5 are AAV1 and AAV9 hybrid capsids⁴⁴.
^‡The following 11 AAV2R585E hexapeptide mutants were included: 463-16000, 465-00700, 465-16000, 467-16000, 479-00009, 573-00780, 581-10000, 583-06000, 585-00080, 593-10000 and 595-10000.
^§The amino-acid positions are provided based on AAV2 VP1.

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