Multi-targeted gene silencing strategies inhibit replication of Canine morbillivirus

BMC Veterinary Research

Table 1 Effects of transiently pENTR/U6-expressed shRNAS on CDV replication

shRNA construct	Log₁₀ reduction value^a and CPE score^b
	pENTR/U6-shRNA (μg)
	0.25			0.5			0.75			1
	qRT-PCR	TCID₅₀	CPE	qRT-PCR	TCID₅₀	CPE	qRT-PCR	TCID₅₀	CPE	qRT-PCR	TCID₅₀	CPE
Ni1	1.35^a	1.17^a	3.0	2.14^b	1.67^b	2.0	2.23^b	3.28^c	1.3	2.65^c	3.28^c	1.0
Ni2	0.71^a	1.22^a	3.0	1.48^b	2.22^b	2.3	1.92^c	3.00^b	1.8	2.53^d	3.00^b	1.0
Ni3	1.12^a	1.28^a	3.0	1.63^b	2.44^b	2.3	2.03^bc	2.78^b	1.3	2.29^c	2.78^b	1.0
Li1	ns	ns	4.0	0.29^a	0.61^a	3.0	0.27^ab	0.50^a	3.0	0.41^b	0.78^a	2.0
Li2	0.26^a	0.50^a	3.0	0.36^ab	0.56^a	3.0	0.39^b	0.50^a	3.0	0.31^ab	0.56^a	2.3
Li3	ns	ns	4.0	0.12^a	ns	3.0	0.17^a	ns	3.0	0.27^b	0.72^a	2.0
SCR	ns	ns	4.0	ns	ns	4.0	ns	ns	4.0	ns	ns	4.0
Ni shRNAs association^c	Ni1 (1.2 μg)			Ni2 (1.2 μg)			Ni3 (1.2 μg)			Ni1 + Ni2 + Ni3 (0.4 μg each)
Ni shRNAs association^c	2.54^b	3.1^a	1.0	1.90^a	3.1^a	1.0	1.92^a	3.1^a	1.0	1.87^a	3.1^a	1.0

^aThe log₁₀ reduction was calculated by subtracting the log₁₀ means of the CDV infectivity in the presence of transfected shRNA constructs from the log₁₀ means of the CDV infectivity in the absence of shRNAs
^bCPE scores were defined as (1) CPE < 25%, (2) CPE between 25 and 50%, (3) CPE between 51 and 75% and (4) CPE > 75%. CPE values correspond to the CPE score means for replicates
^cAssociation of pENTR/U6-shRNAs against CDV nucleoprotein
Values followed by the same lowercase letters in horizontal lines do not differ within each method; p < 0.01 (Tukey’s test)
Ni plasmid shRNA for CDV nucleoprotein mRNA, Li plasmid shRNA for CDV large protein mRNA, SCR plasmid scramble shRNA, ns (not statistically significant): no significant difference between shRNA-treated and untreated CDV infected cells

ISSN: 1746-6148