This document summarizes a gene knockdown product that uses shRNA plasmids to suppress target gene expression in human, mouse, and rat cell lines. It guarantees that at least two of the four shRNA plasmids provided will reduce target gene expression by 70% or more. The plasmids contain antibiotic resistance or GFP markers to select for or enrich stable transfectants for studying long-term or short-term gene suppression effects, respectively. It provides experimental validation and specificity testing to minimize off-target effects.

1. Guaranteed Knock Down for Every Human, Mouse, and Rat Gene
Our experimentally verified shRNA design algorithm ensures the
maximum gene-specificity and RNAi efficacy. We guarantee that
you will see 70% or greater knock down for your targeted gene
with at least two of the four shRNA plasmids.* The availability of
two effective sequences allows you to properly control for nonspecific and off-target effects.
Enrich or Select:
Use Neomycin or Puromycin Resistance for stable
selection and study the long term effects of gene
suppression.
Use GFP Marker for FACS enrichment and the study of
short term effects of gene suppression.
SureSilencing shRNA Plasmid Kit Contents:
4 shRNAs targeting gene-of-interest
1 negative control shRNA
Plasmid Options:
For FACS Sorting:
GFP Marker (Cat. Nos. KX#####G)
For Stable Selection:
Neomycin (Cat. Nos. KX#####N)
Puromyin (Cat. Nos. KX#####P)
Prepare stock of transfection-grade plasmids
by amplification in bacteria.
Transfect shRNA Plasmids into Cell Line of Interest.
Transfect all 4 shRNAs and Negative Control into your cell line in a
six-well culture plate.
Guaranteed!*:
Knock down expression of the target gene by at least
70 percent. Control for non-specific and off target
effects.
Convenient and Cost-Effective:
Use standard plasmid-based and lipid-mediated
transfection methods. Plasmids provide a renewable
source of RNA Interference.
e
ativ
Neg
4
NA
shR
3
NA
shR
2
NA
shR
1
NA
shR
hairpin RNA (shRNA) designs packaged in a plasmid backbone.
3 days
interference. For each gene, we provide four separate short
How It Works
shRNA 1
shRNA 2
shRNA 3
shRNA 4
24 - 48 hours
the expression of every human, mouse, or rat gene by RNA
Negative
If using GFP Marker Plasmid,
Analyze Short Term Effects of Gene Suppression
Enrich by FACS or analyze gene function by fluorescence microscopy.
> 1 day
SureSilencing™ shRNA Plasmids specifically knock down
If using Selection Markers,
Analyze Longer Term Effects of Gene Suppression
100
80
Select for stable transfectants using Neomycin or Puromycin.
60
1 - 2 weeks
- OR-
Guaranteed Knock Down
Verify Suppression of Gene Expression
Using Real-Time PCR
40
hours
Relative Expression Level
(Percent of Control)
RNA InterFerence
SureSilencing™ shRNA Plasmid-Based RNA Interference:
20
0
MAP3K1 ITGB1
MYC
SP1
STAT1 HSPCB
Targeted Gene
SureSilencing™ shRNA Plasmids knock down target gene expression in
transfected cells by at least 70 percent. SureSilencing shRNA Plasmids
with GFP Marker for each of six genes were transfected into HEK-293 cells.
The transfection efficiency was estimated by fluorescence microscopy. The
relative expression level of each target gene, normalized to transfection
efficiency was determined by qRT-PCR and plotted with the standard
deviation of the biological and technical replicates.
Perform biochemical, cell, or molecular biological
assay of gene function

2. Feature Summary
Tested
Experimentally verified design algorithm ensures efficacy.
Specificity
Minimizes off-target effect with the Smith-Waterman specificity search.
Plasmid Backbone
U1 promoter transcribes moderate shRNA amount: minimal off-target effect & toxicity
Bacterial origin of replication and ampicillin-resistance marker: amplify life-time supply
Mammalian Expression Markers:
Neomycin or Puromycin Resistance: select stably transfected cells , study long term effects
GFP Marker: enrich or track transiently transfected cells, study short term effects
Unlike chemically synthesized siRNA, plasmid-based shRNA
provides a renewable source of RNA interference. You can
Applications
Use RNAi in virtually any cell line:
Accommodates both low and high transfection efficiencies.
amplify enough plasmid to complete any number of projects in
your lab. Purchase of SureSilencing™ shRNA plasmids includes
complete shRNA sequence information.
Stable transfections with resistance markers:
Study slow effects using any gene function assay.
Transient transfections GFP-enriched by FACS:
Study quick effects using any gene function assay.
* The SureSilencing™ shRNA Guarantee: At least two of the provided
pre-designed SureSilencing™ shRNA Plasmids are guaranteed to
knock down expression of the targeted gene at the RNA level by at
least 70 percent as measured by real-time qRT-PCR in transfected
cells upon FACS-based enrichment for GFP expression or selection for
neomycin or puromycin resistance as described in the User Manual.
Transient transfections GFP-tracked by
fluorescence microscopy:
Study quick effects using cell-based imaging and gene
function assays.
SureSilencing™ shRNA Plasmid Ordering Guide
Find the shRNA for your genes at:
Cat. No.
KX#####N
KX#####P
KX#####G
www.SABiosciences.com/RNAisearch.php
Description
SureSilencing™ shRNA Plasmid with Neomycin Resistance
SureSilencing™ shRNA Plasmid with Puromycin Resistance
SureSilencing™ shRNA Plasmid with GFP Marker
X = H for Human, M for Mouse, or R for Rat
Related Products:
For Real-Time PCR Verification of Knock Down:
RT2 PCR qPCR Assays: Available for all shRNA target genes and housekeeping genes
RT2 Real-Time™ SYBR Green Master Mix
RT2 First Strand Kit (C-03)
Focus on your Pathway
SABiosciences Corporation
6951 Executive Way
Frederick, MD 21703
www.SABiosciences.com
T 301.682.9200
888.503.3187
F 301.682.7300
888.465.9859