Reporter Gene Plasmid Information

 

We currently sell nine different reporter genes:

  1. Photinus pyralis luciferase (FLuc, pGL4 derived). The brightest and most up to date version of this gene.
  2. Renilla reniformis luciferase (RLuc). Slightly smaller than FLuc and uses a different substrate.
  3. iLumena luciferase, secreted into the supernatant and uses the same substrate as RLuc.
  4. daGFP. A small, bright, synthetic fluorescent gene with similar properties to GFP.
  5. krYFP. A bright synthetic yellow fluorescence protein.
  6. frCFP. A bright synthetic cyan fluorescence protein.
  7. Beta Galactosidase (ß-Gal)
  8. Human Secreted Alkaline Phosphatase (SEAP)
  9. Chloramphenicol Acetyl Transferase (CAT).

 

Plasmid Cloning options: We sell our reporter genes in a range of configurations, including...

  1. In the main multiple cloning site downstream of either yeast, bacterial or mammalian promoters. 
  2. Under control of the Phosphoglycerate Kinase (PGK) promoter for expression in mammalian cells immediately after the main multiple cloning site. In these vectors you can insert an additional gene driven by a second promoter, and both genes use the same poly adenylation signal to terminate expression.
  3. Under the control of an internal ribosome entry site (EMCV IRES or FMDV IRES) for expression in mammalian cells. These can often demonstrate low, or variable activity depending on the sequence, cell line and upstream gene. Total protein content yields of 3-5% of the upstream gene are not uncommon, although 40-60% of cells may register as positive by transfection, depending on the reporter gene and the assay.
  4. Under the control of either the Rous Sarcoma Virus (RSV) promoter (low expression) or the Human Ubiquitin (Ub) promoter (high expression), where the entire expression cassette is inserted away from the main multiple cloning site, flanked by two AscI restriction sites to allow excision or exchange if needed.

 

All of our constructs are designed for broad compatibility and versatile cloning, allowing you easily to produce any derivates you require. If you prefer to outsource the cloning work, we are happy to undertake it rapidly and efficiently. Please contact us to tell us about your project

 


Promoter Expression Plasmid Information

 

Promoter Configurations: The structure of our plasmid vectors means that we are able to  provide any of of the promoter you see in our catalogue in at least 4 different locations in our plasmids. Not all of these configurations will be available on the website so please contact us if you cant see the location you would like. 

 

In our current product range we have a wide range of promoters for expression in different biological systems. These include:

  • Bacterial Promoter - constitutive and inducible
  • Mammalian promoters - endogenous, viral and synthetic
  • Phage promoters for in vitro transcription (T7, SP6 and T3) or inducible bacterial expression (T7)
  • Yeast promoters for low or high expression
  •  

These promoters are typically either flanked by restriction sites, or provided within a pre-designed expression cassette which itself is flanked by resitrction sites. This allows you to move expression cassettes between our different plasmids and make complex expression systems in minimal steps. 

 

To view our different promoter options please see our plasmid products section, or please contact us for further information.


Peptide Tags

We stock the world's largest collection of plasmids encoding functional peptide tags. The tags are positioned automatically at either the N or C terminus when you insert your gene of interest into the MCS, using a range of unique enzyme sites.

We currently offer plasmids containing the following tags, for expression in mammalian, yeast or bacterial cells:Mamma Tag Fig

  1. Histidine (6 His)
  2. Histidine (10 His)
  3. Influenza HA
  4. C-Myc
  5. FLAG
  6. Strep
  7. V5
  8. Glutathione-S-transferase (GST)
  9. Maltose binding protein (MBP)
  10. T7
  11. S-Tag

For more information about these tags see our peptide tag guide.

Protease Cleavage Sites

All of our peptide tags are available with five different protease cleavage sites including: Enterokinase (EKT) / Rhinovirus 3C (PreScission) / Tobacco Etch Virus (TEV)/ Thrombin (Throm)/ Factor Xa. For more information on these enzyme sites, please see our cleavage tag guide.

The plasmid structure shown contains a 6His affinity tag and a V5 epitope tag, both upstream of a TEV cleavage site, all positioned upstream of the MCS. When you insert your gene of interest into the MCS, both the functional tags and the cleavage site will be positioned at the N terminal of the expressed protein. This will allow you to produce your protein, purify and characterise it, and simply remove the tags (using TEV proteolysis) if you wish. We have literally hundreds of variations on this structure, to meet your precise requirements.

If you want a plasmid that provides functional tags alongside a secretory tag (signal peptide), to engineer secretion of your protein product from mammalian cells, please check out the products on our mammalian secretory tags page.

We also provide plasmids encoding reporter gene tags (fusion proteins) including Renilla and Firefly luciferase, Beta Galactosidase, Alkaline Phosphatase, Chloramphenicol Acetyl transferase and a range of fluorescent reporters. Please see our Reporter Fusion Proteins page. 

Please use the Tables below to find the mammalian tag plasmid you require. Alternatively you can search using the Plasmid Search tool, or browse our tags catalogues using the buttons at the bottom of this page. If in doubt, please contact us.

Plasmids Introducing Functional Tags onto Your Gene of Interest

 

 

N Terminal Tag Plasmids in Blue

C Terminal Tag Plasmids in Red

Protease Cleavage Site

For removal of tag after purification or analysis of your protein

 

For expression in mammalian cells

For expression in bacterial cells

 

none

EKT

TEV

FXa

Thrombin

3C

none

EKT

TEV

FXa

Thrombin

3C

 

 

Epitope Tags

V5

OG3422

OG3202

OG3411

OG3194

OG3404

OG3191

OG3400

OG3192

OG3401

OG3196

OG3405

OG3316

OG3178

OG3306

OG3167

OG3297

OG3508

OG3294

OG3505

OG3295

OG3506

OG3300

OG3511

 

Ha

OG3215

OG3425

OG3200

OG3409

OG1127

OG1128

OG1067

OG1068

OG1087

OG1088

OG1145

OG1146

OG3319

OG3181

OG3304

OG3515

OG2774

OG2775

OG2708

OG2709

OG2730

OG2731

OG2792

OG2793

 

C-Myc

OG3214

OG3424

OG3199

OG3408

OG1126

OG1125

OG1065

OG1066

OG1085

OG1086

OG1143

OG1144

OG3318

OG3180

OG3303

OG3514

OG2772

OG2773

OG2706

OG2707

OG2728

OG2729

OG2790

OG2791

 

S-Tag

OG3219

OG3429

OG1163

OG1164

OG1135

OG1136

OG1075

OG1076

OG1095

OG1096

OG1153

OG1154

OG3323

OG3185

OG2812

OG2813

OG2782

OG2783

OG2716

OG2717

OG2738

OG2739

OG2800

OG2801

 

T7

OG3218

OG3428

OG1161

OG1162

OG1133

OG1134

OG1073

OG1074

OG1093

OG1094

OG1151

OG1152

OG3322

OG3184

OG2810

OG2811

OG2780

OG2781

OG2714

OG2715

OG2736

OG2737

OG2798

OG2799

 

FLAG

OG3213

OG3423

OG3198

OG3407

OG1123

OG1124

OG1063

OG1064

OG1083

OG1084

OG1141

OG1142

OG3317

OG3179

OG3302

OG3513

OG2770

OG2771

OG2704

OG2705

OG2726

OG2727

OG2788

OG2789

 

Affinity Tags

MBP

OG3220

OG3430

OG1165

OG1166

OG3195

OG3403

OG1077

OG1078

OG1097

OG1098

OG1155

OG1156

OG3324

OG3186

OG2816

OG2817

OG3298

OG3509

OG2720

OG2721

OG2742

OG2743

OG2804

OG2805

 

GST

OG3216

OG3426

OG3201

OG3410

OG1129

OG1130

OG1069

OG1070

OG1089

OG1090

OG1147

OG1148

OG3320

OG3182

OG3305

OG3516

OG2776

OG2777

OG2710

OG2711

OG2732

OG2733

OG2794

OG2795

 

Strep

OG3217

OG3427

OG1159

OG1160

OG1131

OG1132

OG1071

OG1072

OG1091

OG1092

OG1149

OG1150

OG3321

OG3183

OG2808

OG2809

OG2778

OG2779

OG2712

OG2713

OG2734

OG2735

OG2796

OG2797

 

6His

OG3211

OG3421

OG3197

OG3406

OG1121

OG1122

OG1061

OG1062

OG1081

OG1082

OG1139

OG1140

OG3329

OG3517

OG2814

OG2815

OG3299

OG3510

OG2718

OG2719

OG2740

OG2741

OG2802

OG2803

 

10His

OG3210

OG3420

OG1157

OG1158

OG1119

OG1120

OG1059

OG1060

OG1079

OG1080

OG1137

OG1138

OG3315

OG3177

OG3301

OG3512

OG2768

OG2769

OG2702

OG2703

OG2724

OG2725

OG2786

OG2787

 

Dual

Tags

 

V5 & 6His

OG3203

OG3209

OG3412

OG3419

OG3207

OG3417

OG3204

OG3413

OG3205

OG3415

OG3208

OG3418

 

 

 

 

 

 

 

 

HA & 6His

OG1179

OG1267

OG1180

OG1268

OG1235

OG1236

OG1187

OG1188

OG1203

OG1204

OG1251

OG1252

 

 

 

 

 

 

 

 

c-Myc & 6His

OG1265

OG1177

OG1266

OG1178

OG1233

OG1234

OG1185

OG1186

OG1201

OG1202

OG1249

OG1250

 

 

 

 

 

 

 

 

S-Tag & 6His

OG1171

OG1275

OG1172

OG1276

OG1243

OG1244

OG1195

OG1196

OG1211

OG1212

OG1259

OG1260

 

 

 

 

 

 

 

 

T7 & 6His

 

OG1169

OG1273

OG1170

OG1274

OG1241

OG1242

OG1193

OG1194

OG1209

OG1210

OG1257

OG1258

 

 

 

 

 

 

 

 

FLAG & 6His

OG1175

OG1263

OG1176

OG1264

OG1231

OG1232

OG1183

OG1184

OG1199

OG1200

OG1247

OG1248

 

 

 

 

 

 

 

 

MBP & 6His

OG1173

OG1277

OG1174

OG1278

OG1245

OG1246

OG1197

OG3414

OG1213

OG1214

OG1261

OG1262

 

 

 

 

 

 

 

 

GST & 6His

OG1181

OG1269

OG1182

OG1270

OG1237

OG1238

OG1189

OG1190

OG1205

OG1206

OG1253

OG1254

 

 

 

 

 

 

 

 

Strep & 6His

OG1167

OG1271

OG1168

OG1272

OG1239

OG1240

OG1191

OG1192

OG1207

OG1208

OG1255

OG1256

 

 

 

 

 

 

 


 

Our Secretion Plasmids:

We sell a range of secretory signal peptide plasmids that allow the export of a protein from the cytosol into the secretory pathway. Proteins can exhibit differential levels of successful secrection and often certain signal peptides can cause lower or higher levels when partnered with specific proteins. For this reason we sell 10 signal peptides for secretion in mammalian cells, 10 for secretion in bacterial cells and 6 for secretion from yeast cells. This provides a range of plasmid options to enable the successful secretion of your proteins.
We have had significant success with some of our signal peptides in the past, and for this reason we provide those particular signal peptides in conjunction with a range of other peptide tags, such as His tags and epitope tags. These are generally also available with all of the commonly used protease cleavage tags. Tags that are available in these configurations are:

Mammalian - Human Insulin and BM40
Bacteria - OmpA and PelB
Yeast - Alpha Amylase and Full Length Alpha Factor

 

Cloning into our Plasmids

Our Plasmids provide three cloning options -


  1. Standard restriction enzyme cloning (normally into EcoRI and XhoI)

  2. SnapFusion cloning (amplify your gene to add either BsgI or BseRI at each end and clone in a single seamless step. See the cloning section of each product page for more information and help with primer design. 

  3. Gibson assembly - create small arms of homology to the plasmid and clone in your gene in a single step. 

  4.  

SnapFusion Cloning

SnapFusion is a method that we developed that allows you to insert your gene sequence next to a peptide tag without adding any extra bases to your gene.

N-terminal Peptide Tags - Using SnapFusion on any of our N-terminal peptide tag plasmids leaves a TG overhang in an ATG start codon that is always poistioned immediately after the peptide tag sequence in our plasmids. It also leaves a TA overhang on a TAG codon that is further down the multiple cloning site. This is achieved by cutting the plasmid with either BseRI or BsgI. 

To insert your gene amplify it to add any of the followning sites: AcuI, BpmI, BpuEI, BseRI, BsgI, EciI. Using this webtool can help with the primer design if required. Then simply cut your gene and clone it into the plasmid. If using BsgI or BseRI you can simply mix the PCR product and the plasmid and add the enzyme and ligase. The gene will automatically be inserted into the plasmid and cannot be cut back out. T4 DNA ligase is active in the recommended buffers for BseRI and BsgI.

 

General Information - Signal Peptides

In eukaryotes the signal peptide is a hydrophobic string of amino acids that is recognised by the Signal recognition particle (SRP) in the cytosol of eukaryotic cells. After the signal peptide is produced from a mRNA-ribosome complex, the SRP binds the peptide and stops protein translation. The SRP then shuttles the mRNA/ribosome complex to the rough endoplasmic reticulum where the protein is translated into the lumen of the endoplasmic reticulum. The signal peptide is then cleaved off the protein to produce either a soluble, or membrane tagged (if a transmembrane region is also present), protein in the endoplasmic reticulum. 

Signal peptides contain the sequences that are responsible for their own cleavage. This cleavage point will be highlighted in each individual product data sheet.

In prokaryotes, the most commonly used secretory tags are the OmpA and PelB secretion tags. These signals peptide function similarly to their eukaryotic counterpart, however, because prokaryotes have no internal membranous organelles, and bacteria have either a cell wall (gram-positive) or a second membrane and a cell wall (gram-negative), the protein is normally secreted into the periplasmic space rather than the supernatant.

Please browse our reporter constructs using the buttons below. All of our constructs are designed for broad compatibility and versatile cloning, allowing you easily to produce any derivates you require. As ever, if you prefer to outsource the cloning work, we are happy to undertake it rapidly and efficiently. Please use the 'Build Your Own Secretory Peptide Plasmid' button below to evaluate possible modifications to your chosen plasmids.


What is an Internal Ribosome Entry Site (IRES)?

In a sentence: An IRES is an RNA sequence that forms a complex secondary strcuture that allows the initiation of translation from any position within an mRNA immediately downstream from where the IRES is located.

More Information: 

In order to explain what an IRES is we must first consider their origin and natural function. The most commonly used IRES sequences used in protein expression are derived from the picornavirus family of viruses, which include polioviruses and foot and mouth disease viruses. When these viruses enter cells they are often inflammatory and can induce the expression of genes that will prevent virus gene expression such as interferons. They also, as most viruses do, want to convert the cell into factories for producing their own proteins.

In order to this efficiently, these viruses express proteins that prevent ribosomes from engaging on mRNA molecules inside infected cells. This means that almost all protein production is stopped inside infected cells. However, the virus still needs to produce its own proteins and as such has evolved an RNA sequence that folds in a particular way that allows ribosomes to bind and start protein translation, independent of normal translation routes. This allows the virus to produce its proteins even though all 5’ cap-dependent translation has been inhibited within the cell. This also means that the virus is able to load ribosomes onto an mRNA from any region within an mRNA where the IRES is located. Hence why they are termed ‘internal’ ribosome entry sites.

 

How can we exploit IRES sequences?

For research, it is often desirable to express more than one gene from a stretch of DNA. The difficulty of predicting splice sequences, which would provide the ideal solution to this problem, has traditionally been made this hard to achieve. The observation that some viruses possess sequences that allow the loading of ribosomes for translation from ‘internal’ positions within an mRNA provided a potential solution. By positioning one coding sequence downstream of the 5’ cap/5’UTR in an mRNA, and a second gene downstream of an IRES sequence it is possible to allow the expression of two genes from a single mRNA (see figure below).

 

 

Commonly used sequences

The main IRES sequences used for the expression of exogenous genes are derived from Foot and Mouth Disease virus (FMDV) and Encephalomyocarditis virus (EMCV). The EMCV virus sequence is much more frequently used and consistently delivers slightly higher expression in the cell types we have tested. However, the EMCV virus sequence is longer than the FMDV virus sequence, which can be an important consideration in space constraint expression systems (lentivirus and adenoviruses for example).

 

IRES Expression levels

There is no shortage on literature pertaining to IRES expression levels. At Oxford Genetics we have spent more time on trying to understand IRES expression than almost any other sequence group. This is because they often show low expression levels compared to the upstream genes, and the level of expression varies from cell type to cell type. The following is a very brief conclusion of our findings:


  1. IRES expression is always lower than the upstream gene in vivo if using a strong promoter such as CMV or EF1-Alpha

  2. The position if the start codon of the gene is important but movement either side of the ideal position will normally be tolerable but may give slightly lower expression.

  3. Expression will vary from cell type to cell type and if using cells that are hard to transfect it can be difficult to use selection markers controlled by an IRES. For example, suspension immune cell lines are very difficult to select for with IRES systems.

 

Our current hypotheses and a possible explanation for the variable data in the literature are:

  1. 1: Often experiments are conducted in vivo, but also some published studies use in vitro transcription. Using in vitro transcription it is hard to determine how efficient the 5’ capping of the mRNA has been, therefore making comparisons between upstream gene and downstream gene levels hard to interpret.

  2. It is likely that IRES systems are saturable in vivo. For this reason, so if you produce 100 000 mRNAs from a CMV promoter but the IRES system in a given cell type can only load onto 10 000 mRNAs, the relative expression level from the upstream gene to the downstream gene will be around 10:1. However, if you use a weaker promoter, perhaps SV40, that might produce only 10 000 mRNA copies, the IRES system can load this many and so the ratio is now 1:1 between the upstream and downstream gene. We have not yet conclusively proven this, but it seems to explain the conflicting data in the literature.

  3. Measuring positive cells doesn’t measure protein levels. Often in the literature measurements are made using GFP by FACS. Then data is interpreted on the basis of positive cells. All cells expressing the upstream gene will also be expressing the downstream gene, so there will be a 1:1 ratio of expression. However, this does not mean there is as much of the upstream protein and there is of the downstream protein. In our studies we have focused mainly on total protein yield.  

 

In our studies, using a CMV promoter, the EMCV IRES produces between 10 and 20 fold less protein than the upstream gene, whilst FMDV IRES produces typically 20-30 fold less.


\"gallery

Bacterial Origin Cloning: We currently have four alternative origins of replication for use in bacteria. Our bacterial origins of replication are normally flanked by two SwaI restriction sites. They will function if they are inserted in either orientation between these two sites.

 

Phage and Mammalian Origin Cloning: We also have a range of other origins of replication, including the SV40 origin of replication for mammalian cells and the F1 phage origin of replication that can be used to create single stranded DNA. These are currently available in either the SbfI or PacI sites respectively, however, they can be inserted into other positions on request.

 


We have produced three different types of plasmids containing industry standard transcription termination signals. These include:

  1. Plasmids containing three transcriptional terminators all positioned downstream of the main multiple cloning site (MCS), including the SV40 PolyA signal, RrnG bacterial terminator and the T7 polymerase hair-pin terminator.  This single terminator region provides flexibility when exchanging between mammalian, bacterial and phage systems. This system is present in the vast majority of our plasmids.

  2. Plasmids designed for expression in yeast contain the CYC1 terminator, together with the RrnG bacterial terminator and the T7 polymerase hair-pin terminator.

  3. We also produce plasmids containing a single terminator downstream of the MCS, namely the SV40 polyA mammalian terminator (product OG176), RrnB bacterial terminator (Product OG179), RrnG bacterial terminator (Product OG180) or the T7 polymerase hair-pin terminator (Product OG177).

 

The reason that we stock plasmids that only contain one terminator is because sometimes, despite the increased versatility, researchers may not want unnecessary terminators in their plasmids, for example when size constraints are an issue. 

The button below takes you to the plasmids with single terminators. If you prefer to use a plasmid with triple terminator, we suggest you search using the Plasmid Seach tool since nearly all our plasmids have triple terminators. The Plasmid Search tool will give you access to our full plasmid catalogue.

Finally, please try the 'Design Your Own Plasmid Online' button below, to see what our cloning system can do for you. And remember that, while our plasmids are designed for easy cloning modifications, we are happy to do it for you if you prefer to outsource the cloning work.

 


About  CRISPR 

Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) contribute to a bacterial system for defence against infection from phages, similar to acquired immunity in eukaryotes. Key components of CRISPR have been adapted for use as a tool for genome engineering in a variety of organisms. The editing system consists of two components: CRISPR-associated endonuclease 9 (Cas9) and a guide RNA (gRNA). Cas9 protein creates a double strand DNA break at a site in the genome that is defined by the sequence of a gRNA molecule that is bound to the Cas9 protein. The location at which the Cas9 protein cuts the DNA is defined by the unique sequence of the RNA that is bound to it. The gRNA consists of two sections, a scaffold region required for the RNA to bind to Cas9 and a 20 nucleotide targeting-sequence, which directs Cas9 to the desired cut position in the genome.

 

Cas9 Cutting Process

First, a DNA molecule is introduced into a cell that encodes the Cas9 protein and also encodes an RNA molecule that has both the scaffold sequence and a sequence that will bind to location on the genome to be cut. Following transcription and translation the Cas9 protein binds to the scaffold section of the gRNA. This forms a gRNA-Cas9 complex causing a conformational change in the Cas9 protein enabling the RNA-protein complex to bind to double stranded DNA at loci defined by the guide RNA. This guide must contain the sequence NGG, the Protospacer Adjacent Motif (PAM) at the 3' end (see figure 1 'PAM' section).  It is important to note that the NGG PAM sequence is not in the guide RNA molecule, but must be in the genome to allow cleavage.

Whether the PAM bound-Cas9 cleaves the DNA strands depends on base pairing between one of the genomic DNA strands and the targeting region of the gRNA (figure 1). Base pairing begins at the 3’ end of the gRNA targeting region and propagates along towards the 5’ terminus. A 100% match will lead to efficient cleavage, however, Cas9 tolerates up to 7 mismatches toward the 5’ end of the gRNA. There must be no mismatches in the 11 base pairs (bp) preceding the PAM site (Cong et al., 2013). This is an important factor when considering potential off-target binding sites. Online tools are available to help minimise off-target binding (please see below). For more information on CRISPR experimental design please see the following review (Graham & Root, 2015).

 

\"how

 

Figure 1: Diagram of the genomic DNA, gRNA and Cas9 complex. The NGG PAM site is not present in the gRNA but must be present in the genomic target sequence. The Cas9-gRNA riboprotein complex binds the genomic DNA and base pairing between the targeting-region and complementary strand leads to cleavage of the genomic DNA.

Once Cas9 has cleaved the genomic DNA, usually 2-4 bp upstream the PAM site, the resulting double stranded break (DSB) is repaired by one of two cellular pathways:


  1. An efficient but error prone pathway called non-homologous end joining (NHEJ) and 

  2. A less efficient but more accurate pathway called homology directed repair (HDR), which requires an intact DNA repair template.

 

Disrupting a Gene Sequence with CRISPR & Cas9

If the double strand break is repaired by NHEJ, this often results in short insertions or deletions (indels) at the cleavage site. This is the mode of repair used if no homologous repair template is provided. NHEJ creates short insertions or deletions where the break occurred. If the cleavage site is within a coding region, and if that mutation causes a frame-shift, it will effectively disrupt / silence that gene at the genomic level. The CRISPR system can be used to disrupt genes and generate knock-outs cells by co-expressing a gRNA specific to the target gene with Cas9 (see figure 1). The target sequence must be immediately upstream a Protospacer Adjacent Motif (PAM). The sequence of the PAM site is NGG. The target sequence must be immediately upstream a PAM (NGG) site.

 

Oxford Genetics’ Vectors

At Oxford Genetics we offer vectors containing cassettes for expressing Cas9 plus a gRNA of choice. These may be in the same vector, which simplifies experimental set up and improves co-transfection efficiency; or in separate vectors, which allows more control over the relative dosage of Cas9 versus gRNA. We also offer an adapter to allow expression of two different gRNAs from the same vector. This may be useful, for instance, for disrupting two genes simultaneously or making large deletions. We offer Cas9 expression vectors with a variety of different promoters to tailor Cas9 expression to the desired cell-type and/or required expression level. We offer a range of expression vectors where Cas9 is co-expressed with a marker gene (either GFP or puromycin resistance) by means of an internal ribosome entry site (IRES).

 

\"CRISPR

 

Figure 4: This image show some of the more popular sequences and plasmid variants from our CRISPR plasmid platform. Our plasmid collection includes over 2000 unique sequences of functional components (peptide tags, reporter genes, selection genes). These can all be inserted into our CRISPR plasmids through our custom cloning services. Contact our team for more information. 

 

References

Cong, L., Ran, F. A., Cox, D., Lin, S., Barretto, R., Habib, N., … Zhang, F. (2013). Multiplex genome engineering using CRISPR/Cas systems. Science (New York, N.Y.), 339(6121), 819–23. 


Graham, D. B., & Root, D. E. (2015). Resources for the design of CRISPR gene editing experiments. Genome Biology, 16(1), 260.