I am looking to use shRNA to inhibit Moesin expression. I have had people advise me that my initial MOI should be low as 'less is more' and 'a little goes a long way' in terms of siRNA. I was wondering if you could elaborate on this for me and explain why my initial MOI should be low.

Name: Moesin Overexpression Lysate
Brand: Novus Biologicals
SKU: NBL1-13334
Price: 709 USD

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The reason for a low MOI is most likely because RNAi is a very strong and efficient technique. Wikipedia does a good job of explaining RNA interference. However, I would imagine that in a cell, there will be at most 1-2 copies of the gene mRNA present at any given time, unless you're dealing with a highly expressed protein such as Actin, where I would imagine silencing Actin would be lethal to the cell. I can imagine a few reasons to not use too much siRNA. First, it is expensive, so you don't want to waste it. Second, using too much would cause there to be a lot of non-translatable RNA present in the cell, which could trigger an immune response, as the presence of uncapped RNAs can indicate presence of a virus and one of the TLRs may respond to this.

At	=	A. thaliana	SyHa	=	Golden Syrian Hamster
All	=	All Species	Ha	=	Hamster
An	=	Animal	Hu	=	Human
ArHa	=	Armenian Hamster	I	=	Insect
Av	=	Avian	Ll	=	Llama
Bb	=	Baboon	Ma	=	Mammal
Ba	=	Bacteria	Mk	=	Monkey
Bv	=	Bovine	Mu	=	Mouse
Ca	=	Canine	Multi	=	Multi-species
Ce	=	C. Elegans	NA	=	Non-species specific
Ch	=	Chicken	Or	=	Orangutan
ChHa	=	Chinese Hamster	Pl	=	Plant
Cm	=	Cynomolgus Monkey	Pm	=	Primate
Do	=	Donkey	Po	=	Porcine
Dr	=	Drosophilia	Rb	=	Rabbit
Ec	=	E. Coli	Rt	=	Rat
Eq	=	Equine	RM	=	Rhesus Macaque
Fe	=	Feline	Sh	=	Sheep
Ft	=	Ferret	Vb	=	Vertebrate
Fi	=	Fish	Vi	=	Virus
Fu	=	Fungi	Xp	=	Xenopus
Ge	=	Gerbil	Ye	=	Yeast
GP	=	Guinea Pig	Ze	=	Zebrafish
Gt	=	Goat

Reactivity	HuSpecies Glossary
Applications	WB

Dilutions	Western Blot
Application Notes	This product is intended for use as a positive control in Western Blot. Overexpression of the target protein was confirmed using an antibody to DDK (FLAG) epitope tag (NBP1-71705) present on the protein construct. Each vial of cell lysate contains 100ug of total protein which should be sufficient for 20-50 reactions. Depending on over-expression level, antibody affinity and detection system, some lysates can go as low as 0.1 ug per load. We recommend starting with 5ug of cell lysate. Add an equal amount of cell lysate and 2X SDS Sample buffer and boil the SDS samples for 10 minutes before loading.
Theoretical MW	67.6 kDa. Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Storage	Store at -80C. Avoid freeze-thaw cycles.
Buffer	RIPA buffer

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