Table 1:

CRISPR/Cas9-mediated HDR in Primary B cells

	Johnson (17)	Hung (18)	Pesch (19)	Luo (20)	Voss (21)	Greiner (22)	Hartweger (23)	Moffett (24)	Nahmad (25)	Huang (26)
Modified Cell Type	Human B cells	Human B cells	Murine B cells	Human B cells	Human B cells	Human B cells	Human and murine B cells	Human and murine B cells	Human and murine B cells	Murine B cells
Pre-transduction activation	CD40L and IL4	CD40L, CpG, IL2/10/15	IL4, BAFF, CD40L feeder cells	CD40L, IL2/10/15, and CpG	CD40L and IL4	CellXVivo Human B Cell Expansion Kit	Anti-TLR4 Ab (RP105)	hCD40L, IL2/10/15, and CpG (h) mCD40L + IL4 (m)	Anti-TLR4 Ab (RP105), IL7 (h) LPS (m)	LPS, CpG
Delivery Vehicle	AAV6 + RNPs	AAV6 + RNPs	Linear dsDNA/AAVDJ +RNPs	BaEV-pseudtotyped Integrase-deficient LV	plasmid	ssDNA + RNPs	ssDNA + RNPS	AAV6 + RNPs	AAV6 + RNPs (h) AAVDJ + RNPs (m)	Plasmid / AAV2 RNPs
Integration Site	AAVS1 locus	CCR5 locus	Rosa26 locus	GAPDH locus	IgH locus	IgH and IgK loci	IgH locus	IgH locus	IgH locus	IgH and IgK loci
Cutting efficiency	Not evaluated for AAVS1 locus	43–84%	Not evaluated	48.3%	Not evaluated	not quantified for Ig loci	64% (h) 80% (m)	72% (h) 71% (m)	29% (h) 30% (m)	55–80%
Inserted sequences	EGFP	FIX and BAFF	CBCR	αPD1-CD90	VH	HC/nanobody (into IgH) KC (into IgK)	KC-P2A-VH	KC-linker-VH	KC-linker/P2A-VH	KC-P2A-VH or VH (into IgH) VL (into IgK)
HDR efficiency	25%	10–40%	0.3%	21.6%	0.21%	Approx. 15–35%	0.4% (h) 4% (m)	10–60% (h) 10–24% (m)	11% (h) 10–84% (m)	10% (HC)
Promoter	endogenous PPP1R12C	donor derived MND	donor derived CMV	endogenous GAPDH	endogenous HC V-gene	endogenous HC V-gene	donor derived HC V-gene	donor derived HC V-gene	HC V-gene promoter or splice acceptor	donor derived HC V-gene
Target	N/A	Hemophilia/ B cell engraftment	HEL	PD-1	HIV	TNF-α	HIV	RSV, HIV, Influenza, and EBV	HIV	HIV
Secretion in immunocompetent mice	N/A	N/A	N/A	N/A	N/A	N/A	7 days	7–12 days	33 days	252 days
Secretion in NSG mice	N/A	21 days	N/A	150 days	N/A	N/A	N/A	72 days	N/A	N/A