Novel promoter, lentiviral vector and methods and uses thereof

Abstract

Disclosed herein is a promoter sequence comprising at least a portion of a ABCB4 promoter sequence or reverse complement thereof, wherein the splice sites of the ABCB4 promoter are mutated, and lentiviral vector sequences comprising said promoter sequences operably linked to a transgene. Preferably, the promoter sequence and transgene are present in a reverse expression cassette in reverse orientation to the direction to the lentiviral vector sequence. When a transgene for ABCB4 is used, the lentiviral vector can be used in the treatment of PFIC3.

Description

[0001] Novel Promoter, Lentiviral Vector and Methods and Uses Thereof

[0002] Background

[0003] Progressive familial intrahepatic cholestasis type 3 (PFIC3) is just one of a group of PFIC disorders with an incidence of approximately 1:50,000 births. PFIC3 is a life-limiting autosomal recessive disease caused by bi-allelic loss-of-function mutations in ATP-binding cassette subfamily B member 4 (ABCB4; OMIM: 171060) gene encoding hepatocyte apical membrane phosphatidylcholine transporter called multidrug resistance protein type 3 (MDR3). Phosphatidylcholine is normally secreted into bile by MDR3 and is required for protection of bile duct cells from bile acid- and cholesterol crystals- associated injury. Absence of ABCB4 leads to progressive liver fibrosis, cirrhosis and failure. UK patients with PFIC3 are typically diagnosed in infancy upon 1) observation of jaundice (abnormal yellow skin pigmentation), 2) detection of increased conjugated bilirubin levels in blood and 3) detection of bi-allelic pathogenic changes in ABCB4 using gene panels.

[0004] Without any curative treatment, PFIC3 patients often require a liver transplantation. While both mRNA therapy (Wei G et al, J Hepatol. 2021 Jun 1;74(6): 1416-28) and Adeno Associated Virus (AAV) therapy (Aronson SJ et al. J Hepatol. 2019 Jul 1;71(1): 153-62), have been developed for this disease, both of these therapies have a set of limitations that hinder their development to clinical stages. The half-life of most mRNA therapies is relatively short and frequent drug administration would be required to preserve the therapeutic effect. While liver-targeted AAV gene therapy products have been approved for treatment of adults with haemophilia A and B, there are several limitations for AAV based gene therapy: The episomal nature of the AAV means that its effect in children with a growing liver is short-lived because of dilution / loss of non-integrated AAV genomes. Also, pre-existing immunity to AAV capsid means that 50% of the patients may not benefit from the gene therapy. Thus, it is clear that improved approaches are needed for gene therapy for liver diseases, both generally and specifically for the treatment of PFIC3.

[0005] While in vivo lentiviral gene therapy has previously been considered for the treatment of PFIC3 and other liver diseases, lentiviral therapeutic approaches are often hindered by the low concentration of lentiviral vectors that can be obtained. Initial lentiviral vectors tested allowed expression of therapeutic transgenes during vector production in packaging cells leading to disruption of the vector packaging process and low titres. Overexpression of therapeutic transgene in target cells, meaning treatments are too strong, is also sometimes encountered. The scale of the challenge has been previously reported in the literature (see van Til NP, Heutinck KM, van der Rijt R, Paulusma CC, van Wijland M, Markusic DM, et al. Alteration of viral lipid composition by expression of the phospholipid floppase ABCB4 reduces HIV vectorinfectivity. Retrovirology. 2008;5:1-9), where it has been said that no one would be able to produce an effective gene therapy for PFIC3. Improved approaches were therefore needed.

[0006] Summary of Invention

[0007] In a first aspect of the present invention, there is provided a promoter sequence comprising at least a portion of a ABCB4 promoter sequence or reverse complement thereof, wherein the splice sites of the ABCB4 promoter are mutated.

[0008] In a second aspect of the present invention, there is a lentiviral vector sequence comprising the promoter sequence of the first aspect, which is operably linked with a transgene.

[0009] In a third aspect of the present invention, and an embodiment of the second aspect of the present invention, there is provided a lentiviral vector sequence comprising the promoter sequence of the first aspect which is operably linked with a transgene, wherein the promoter sequence and the transgene are in reverse orientation to the direction of the lentiviral vector sequence (i.e., the genome of the lentiviral vector). In other words, the lentiviral vector comprises a reverse expression cassette comprising the promoter sequence of the first aspect which is operably linked with a transgene.

[0010] In a fourth aspect of the present invention, there is provided a lentiviral vector comprising the lentiviral vector sequence of the second or third aspect.

[0011] In a fifth aspect of the present invention there is provided a composition comprising the lentiviral vector sequence according to the second or third aspect, or the lentiviral vector of the fourth aspect, and a pharmaceutically acceptable carrier.

[0012] In a sixth aspect of the present invention, there is provided the lentiviral vector sequence of the second or third aspect, the lentiviral vector of the fourth aspect, or the composition of the fifth aspect, for use in therapy. Also disclosed herein is use of the lentiviral vector sequence of the second or third aspect, or the lentiviral vector of the fourth aspect, in the manufacture of a medicament.

[0013] In a seventh aspect of the present invention, there is provided the lentiviral vector sequence of the second or third aspect, the lentiviral vector of the fourth aspect, or the composition of the fifth aspect, wherein the transgene encodes for a liver transmembrane protein, more particularly, encodes for an ATP-binding cassette transporter, and even more particularlyencodes for ABCB4, for use in the treatment of a liver disease. Also disclosed herein, is a method of treating a liver disease, wherein the method of treatment comprises administering to a subject a therapeutically effective amount of the lentiviral vector sequence according to the second or third aspect, or the lentiviral vector of the fourth aspect, wherein the transgene encodes for a liver transmembrane protein, more particularly, encodes for an ATP-binding cassette transporter, and more particularly encodes for ABCB4. Also disclosed herein is use of the lentiviral vector sequence of the second or third aspect, or the lentiviral vector of the fourth aspect, in the manufacture of a medicament for the treatment of liver disease, wherein the transgene encodes for a liver transmembrane protein, more particularly, encodes for an ATP-binding cassette transporter, and even more particularly encodes for ABCB4.

[0014] In an eighth aspect of the present invention, there is provided the lentiviral vector sequence of the second or third aspect, the lentiviral vector of the fourth aspect, or the composition of the fifth aspect, wherein the transgene encodes for a liver transmembrane protein and / or encodes for an ATP-binding cassette transporter, more particularly encodes for ABCB4, for use in the treatment of cholestasis. Also disclosed herein is a method of treating cholestasis, wherein the method of treatment comprises administering to a subject a therapeutically effective amount of the lentiviral vector sequence according to the second or third aspect, or the lentiviral vector of the fourth aspect, wherein the transgene encodes for a liver transmembrane protein and / or encodes for an ATP-binding cassette transporter, and more particularly encodes for ABCB4. Also disclosed herein is use of the lentiviral vector sequence of the second or third aspect, or the lentiviral vector of the fourth aspect, in the manufacture of a medicament for the treatment of cholestasis, wherein the transgene encodes for a liver transmembrane protein, more particularly, encodes for an ATP-binding cassette transporter, and even more particularly encodes for ABCB4.

[0015] In a ninth aspect of the present invention, there is provided the lentiviral vector sequence of the second or third aspect, the lentiviral vector of the fourth aspect, or the composition of the fifth aspect, wherein the transgene encodes for a liver transmembrane protein and / or encodes for an ATP-binding cassette transporter, particularly encodes for ABCB4, for use in the treatment of progressive familiar intrahepatic cholestasis (PFIC), preferably PFIC3. Also disclosed herein is a method of treating progressive familiar intrahepatic cholestasis (PFIC), preferably PFIC3, wherein the method of treatment comprises administering to a subject a therapeutically effective amount of the lentiviral vector sequence according to the second or third aspect, or the lentiviral vector of the fourth aspect, wherein the transgene encodes for a liver transmembrane protein, and / or an ATP-binding cassette transporter, and more particularly encodes for ABCB4. Also disclosed herein is use of the lentiviral vector sequence of the secondor third aspect, or the lentivirus vector of the fourth aspect, in the manufacture of a medicament for the treatment of progressive familiar intrahepatic cholestasis (PFIC), preferably PFIC3, wherein the transgene encodes for a liver transmembrane protein, more particularly, encodes for an ATP-binding cassette transporter, and even more particularly encodes for ABCB4.

[0016] In a tenth aspect of the present invention, there is provided the lentiviral vector sequence of the second or third aspect, the lentivirus vector of the fourth aspect, or the composition of the fifth aspect, wherein the transgene encodes for ABCB4, for use in the treatment of disease associated with mutated ABCB4 and / or aberrant expression of ABCB4. Also disclosed herein is a method of treating a disease associated with mutated ABCB4 and / or aberrant expression of ABCB4, wherein the method of treatment comprises administering to a subject a therapeutically effective amount of the lentiviral vector sequence according to the second or third aspect, or the lentiviral vector of the fourth aspect, wherein the transgene is ABCB4. The disease associated with mutated ABCB4 and / or aberrant expression of ABCB4 may be selected from PFIC3, intrahepatic cholestasis of pregnancy type 3 (ICP3), or gallbladder disease 1 (GBD1) or low phospholipid associated cholelithiasis. Also disclosed herein is use of the lentiviral vector sequence of the second or third aspect, or the lentivirus vector of the fourth aspect, in the manufacture of a medicament for the treatment of a disease associated with mutated ABCB4 and / or aberrant expression of ABCB4, wherein the transgene encodes for ABC B4.

[0017] In a tenth aspect of the present invention, there is provided a host cell comprising the lentiviral vector sequence of the second or third aspect, or the lentiviral vector of the fourth aspect. The host cell may be a hepatocyte cell.

[0018] In an eleventh aspect of the present invention, is the use of the promoter sequence as defined in the first aspect, or the lentiviral vector sequence as defined in the second or third aspect, to improve lentiviral titre. Also defined herein is a method of improving the titre of a lentiviral vector sequence, comprising using the promoter sequence of the first aspect, preferably using the lentiviral vector sequence of the third aspect (i.e., comprising a reverse expression cassette).

[0019] In a twelfth aspect of the present invention, there is provided a plasmid comprising the lentiviral vector sequence of the second of third aspect.

[0020] The present invention provides one or more advantages over the prior art.The present invention provides an improved novel liver-specific promoter sequence and lentiviral vector sequences comprising the novel promoter. The promoter sequence and lentiviral vector sequence comprising the novel promoter sequence promote more physiological levels of gene expression and appropriate localisation of transgene expression in hepatocytes, while also significantly increasing lentiviral vector titre overall (i.e., > 10E+7vg / ml). In contrast, comparative lentiviral constructs comprising a known LP1 promoter, commonly used in liver gene therapies, resulted in overexpression of the transgene leading to target cell depolarisation, and was associated with very poor titres of < 10E+4vg / ml.

[0021] Notably, the present inventors also found that directly replacing LP1 with the novel promoter of the invention resulted in expression of the ABCB4 transgene at levels closer to wild-type in a PFIC3 cell model and expression of transgenes in the liver post / n vivo injections. The novel promoter also improves target cell polarisation as compared to the comparative known vector LP1. While these results are demonstrated for a lentivirus vector expressing the ABCB4 transgene, it is envisaged that the novel promoter could be utilised in any other liver-directed gene therapy, particularly with transgenes relevant for other disorders that have similar titre issues.

[0022] The novel promoter can be engineered from a naturally occurring ABCB4 promoter or portion thereof, (e.g., the human ABCB4 promoter, such as SEQ ID NO: 23 or SEQ ID NO: 65) but wherein the splice acceptor and donor sites have been mutated (e.g., SEQ ID NO: 1 or 68 respectively). As such, the sequences are structurally distinguished from naturally occurring sequences (e.g., in the human genome). Mutation of these splice sites is paramount for the safety of any lentiviral therapeutic, since the presence of splice sites in the lentiviral genome can lead to aberrant splicing of the gene in which the vector is integrated. Additionally, the mutation of active splice sites is important, since their presence could potentially lead to the removal of the intronic region containing parts of the promoter from the ssRNA genome of the lentiviral vector leading to a truncated promoter in the vector genome that is integrated in the host DNA.

[0023] In a preferred embodiment of the second aspect, and according to the third aspect of the present invention, the promoter of the present invention is preferably used within a lentiviral vector sequence having a reverse expression cassette. This importantly was found to prevent expression of the transgene in producer cells, while allowing transgene expression in hepatocyte cells. This improved the titre even further (>1E+8vg / ml). Such a result was particularly unexpected since flipping the cassette has previously been associated with providing decreased titre. (See, e.g., Jiaying Han, et al “Improved lentiviral vector titers froma multi-gene knockout packaging line, Molecular Therapy - Oncolytics, Volume 23, 2021, P

[0024]

[0025] ages 582-592, ISSN 2372-7705, and Jiaying Han et al “ / 3-Globin Lentiviral Vectors Have Reduced Titers due to Incomplete Vector RNA Genomes and Lowered Virion Production, Stem Cell Reports" Volume 16, Issue 1, 2021, Pages 198-211, ISSN 2213-6711, https: / / doi.org / 10.1016 / stemcr.2020.10.007)

[0026] While this preferred novel backbone is demonstrated in a lentivirus vector sequence expressing the ABCB4 transgene, it is envisaged that this approach and lentiviral backbone could be utilised for other transgenes and liver-directed or hepatocyte derived gene therapies.

[0027] Lentiviral vectors and lentiviral vector sequences in accordance with the present invention were shown to be effective in the treatment of PFIC3 model mice fed with a cholestatic diet. Mice showed a significant improvement when treated with the lentiviral vector of the invention as compared to a control.

[0028] Embodiments and further aspects of the present invention will now be described.

[0029] In some embodiments of all aspects, the promoter sequence corresponds to a portion of the human ABCB4 promoter sequence or reverse complement thereof. In some embodiments, at least a portion of the promoter sequence comprises at least 95 % sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity, or is identical to, SEQ ID NO: 1, wherein the splice sites are mutated (i.e., the sequence is not SEQ ID NO: 23, i.e., the naturally occurring sequence). In some embodiments, at least a portion of the promoter sequence comprises at least 95 % sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity, or is identical to, SEQ ID NO: 68, wherein the splice sites are mutated (i.e., the sequence is not SEQ ID NO: 65, i.e., the naturally occurring sequence) or the reverse complement thereof.

[0030] In some embodiments, at least a portion of the promoter sequence comprises at least 95 % sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity, or is identical to, SEQ ID NO: 2, wherein the splices sites are mutated (i.e., the sequence is not SEQ ID NO: 24). While it is anticipated that alternative ABCB4 promoter sequences could be used, these sequences are representative of the ABCB4 promoter sequences that are used in the application examples.

[0031] SEQ ID NO: 2 is the reverse complement of SEQ ID NO:1. SEQ ID NO: 2 is utilised in the lentiviral vector sequence where the promoter sequence and the transgene are in reverseorientation to the direction of transcription of the lentiviral vector sequence (i.e., according to the third aspect). SEQ ID NO: 1 is utilised in the lentiviral vector sequence where the promoter sequence and the transgene are in the same orientation to the direction of the transcription of the lentiviral vector sequence. SEQ ID NO: 1 may be referred to as LPa promoter herein, while SEQ ID NO: 2 may be referred to as a reverse complement of the LPa promoter.

[0032] In some embodiments, at least a portion of the promoter sequence comprises at least 95 % sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity, or is identical to, SEQ ID NO: 3. In some embodiments, at least a portion of the promoter sequence comprises at least 95 % sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity, or is identical to, SEQ ID NO: 4.

[0033] SEQ ID NO: 4 is the reverse complement of SEQ ID NO:3. SEQ ID NO: 4 is utilised in the lentiviral vector sequence where the promoter sequence and the transgene are in reverse orientation to the direction of transcription of the lentiviral vector sequence (i.e., in the third aspect of the invention). SEQ ID NO: 3 is utilised in the lentiviral vector sequence where the promoter sequence and the transgene are in the same orientation to the direction of the transcription of the lentiviral vector sequence. These are LPa sequences where both the splice sites of the portion of the human ABCB4 promoter have been mutated to CC in SEQ ID NO: 3, or GG in SEQ ID NO: 4, although it is envisaged that different splice mutations may be used. These are the promoter sequences used in the application Examples.

[0034] In preferred embodiments of the first, second and third aspects, the promoter or lentiviral vector further comprises an enhancer sequence or enhancer region, or the reverse complement thereof. In some embodiments the enhancer is an HCR enhancer, preferably as defined in SEQ ID NO: 5 or a reverse complement thereof. In some embodiments, the HCR enhancer has a sequence which has at least 95% sequence identity, at least 96% sequence identity, at least 97% sequence identity, at least 98% sequence identity, or at least 99% sequence identity, or is identical to, SEQ ID NO: 5 or the reverse complement thereof. The reverse complement of SEQ ID NO: 5 may be used where the promoter sequence and the transgene are in reverse orientation to the direction of transcription of the lentiviral vector sequence (i.e., lentiviral vector genome). This is so that it can promote the expression of the reverse complement transgene in target cells.In some embodiments, the promoter or lentiviral vector sequence comprises a sequence with at least 95% sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity, or is identical to SEQ ID NO: 6 or SEQ ID NO: 7. SEQ ID NO: 6 defines the novel LPa promoter sequence directly coupled to a HCR enhancer sequence, and SEQ ID NO: 7 defines the reverse complement thereof. This is expected to promote stable expression.

[0035] In some embodiments of the second and third aspects, the transgene encodes for a liver protein. In some embodiments of the second and third aspects, the transgene encodes for a transmembrane protein, preferably a live transmembrane protein. In some embodiments, the transgene encodes for an ATP binding cassette transporter protein. This is beneficial since the lentiviral vectors of the invention are expressed in the liver and have good tissue specificity.

[0036] In some embodiments of the second and third aspects, the transgene encodes for ABCB4. This is the transgene expressed in the application examples. Lentiviral vectors of the invention encoding for ABCB4 provide an improved therapy for the treatment of PFIC3. As stated above, patients with PFIC3 typically require a liver transplantation without corrective treatment. Alternative therapies, such as AAV gene therapy, are unlikely to provide long-term correction because of the need to treat infants with a rapidly growing liver, and the non-integrating nature of the AAV leads to loss of the gene therapy effect as the hepatocytes divide. In vivo lentiviral gene therapy is therefore a more effective strategy as it allows integration of the ABCB4 into hepatocyte genome and adequate protein expression. In contrast, lentiviral vectors are replication-defective ssRNA vectors containing a transgene expression cassette of choice, packaged in a minimal set of capsid and enzymatic proteins of the parental virus. They have efficient liver transduction due to VSVg pseudotyping, up to 12kb packaging capacity and provide long-term transgene expression due to (i) genomic integration, (ii) low pre-existing immunity to lentivectors and (iii) an excellent safety profile. As shown in the application examples, vectors encoding for ABCB4 have high lentiviral titre, along with other advantages.

[0037] In some embodiments of the second and third aspects, the transgene encodes human ABCB4, i.e., wild-type human ABCB4. In an embodiment, the transgene comprises at least 95% sequence identity, or at least 96% identity, or at least 97% identity, or at least 98% identity, or at least 99% sequence identity, or is identical to SEQ ID NO: 8 or the reverse complement thereof. SEQ ID NO: 8 is the reverse complement of the wild-type ABCB4 transgene and may be used where the promoter sequence and the transgene are in reverse orientation to the direction of transcription of lentiviral vector sequence (i.e., lentiviral vector genome). The reverse complement of SEQ ID NO: 8 may be used where the promoter sequence and thetransgene are in the same orientation to the direction of transcription of the lentiviral vector sequence (i.e., lentiviral vector genome). In some examples, vectors comprising wild type ABCB4 showed higher titre compared to codon-optimised ABCB4.

[0038] In an embodiment, the transgene coding sequence has at least 95% sequence identity, or at least 96%, or at least 97%, or at least 98%, or at least 99% sequence identity, or is identical to SEQ ID NO: 43. SEQ ID NO: 43 is the coding sequence (CDS) of wtABCB4.

[0039] In some embodiments of the second and third aspects, the transgene is a codon-optimised ABCB4. In some embodiments, the transgene comprises at least 95% sequence identity, or at least 96% identity, or at least 97% identity, or at least 98% identity, or at least 99% sequence identity, or is identical to SEQ ID NO: 9 or the reverse complement thereof. The reverse complement of SEQ ID NO: 9 may be used where the promoter sequence and the transgene are in the same orientation to the direction of transcription of lentiviral vector sequence (i.e., lentiviral vector genome) In an embodiment, the transgene coding sequence has at least 95% sequence identity, or at least 96%, or at least 97%, or at least 98%, or at least 99% sequence identity, or is identical to SEQ ID NO: 44. SEQ ID NO: 44 is the coding sequence (CDS) of a codon-optimised ABCB4.

[0040] In some embodiments of the second and third aspects, the transgene is ABCB11. Lentiviral vectors of the invention encoding for ABCB11 provide therapy for the treatment of PFIC2.

[0041] In some embodiments of the second and third aspects, the transgene is a codon-optimised ABCB11. In some embodiments, the transgene comprises at least 95% sequence identity, or at least 96% identity, or at least 97% identity, or at least 98% identity, or at least 99% sequence identity, or is identical to SEQ ID NO: 72 or the reverse complement thereof. The reverse complement of SEQ ID NO: 72 may be used where the promoter sequence and the transgene are in the same orientation to the direction of transcription of lentiviral vector sequence (i.e., lentiviral vector genome).

[0042] In some embodiments of the second and third aspects, the lentiviral vector sequence further comprises one or more miRNA binding sequence, preferably one or more repeat miRNA sequences. In some embodiments, the lentiviral vector sequence comprises one or more of miR-142 and / or miR-126-3pT binding sequences. In some embodiments, the lentiviral vector sequence comprises a sequence with at least 95% sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or atleast 99% identity, or is identical to any one of SEQ ID NO: 10-14 or a reverse complement thereof.

[0043] In some embodiments of the second and third aspects, the lentiviral vector sequence is based on human immunodeficiency virus (HIV) preferably HIV-1.

[0044] In some embodiments of the second and third aspects, the lentiviral vector sequence comprises or has an LTR-LTR sequence that has at least 95% sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity, or is identical to SEQ ID NO: 16 or SEQ ID NO: 15 or SEQ ID NO: 71.

[0045] The present invention provides a novel lentiviral gene therapy that carries a transgene, e.g., the human ABCB4 gene, under the control of a hepatocyte specific promoter, along with a novel lentivirus promoter sequence. The lentivirus vectors disclosed herein can be used for the treatment of liver-related diseases and cholestasis, e.g., PFIC3 for which there are no other effective corrective therapeutic strategies currently available. The lentivirus vectors described herein is supported in at least by a mouse model of PFIC3 which are fed a cholestatic diet, where mice show improvement after vector treatment. As a lentiviral gene therapy, the therapy of the present invention can overcome the above-mentioned challenges described for other gene therapies, including AAV vectors.

[0046] Brief Description of Figures

[0047] Figure 1 shows graphical maps of the different types of lentiviral backbones used in the Examples. (A) Version 1 Backbone - A comparative vector CCL. LP1. Transgene (e.g., CCL. LP1. coABCB4) containing the known LP1 promoter (SEQ ID NO: 42). (B) Version 2 backbone - An example lentiviral vector CCL. LPx. Transgene (e.g., CCL. LPa. coABCB4 and CCL. LPa. coABCB4) containing a novel LPa promoter (SEQ ID NO: 3, with splice sites mutated to CC) (C) Version 3.1 backbone - An example lentiviral vector containing a reverse transgene expression cassette and the reverse complement of the novel LPa promoter (SEQ ID NO: 4). This example vector also comprises a reverse complement of bGH poly(A) signal (SEQ ID NO: 21), and 4x miR-126-3pT binding site and 4x miR142 binding site region (SEQ ID NO: 14). (D) Version 3.2 backbone - an example lentiviral vector (e.g., miR-coA4-LPaH or miR-wtA4-LPaH) which is identical to (C) but additionally contains the novel LPa promoter in combination with HCR enhancer (SEQ ID NO: 5). All constructs contain: SIN 5’ LTR_truncated 5' long terminal repeat (LTR) from HIV-1 (SEQ ID NO: 18), HIV-1

[0048]

[0049] packaging signal of humanimmunodeficiency virus type 1 (SEQ ID NO: 19), cPPT / CTS_ central polypurine tract and central termination sequence of HIV-1 (SEQ ID NO: 20), RRE_ Rev response element of HIV-1 (SEQ ID NO: 46), 3' LTR (AU3)_ self-inactivating 3' long terminal repeat (LTR) from HIV-1 (SEQ ID NO: 22) and the WPRE_ woodchuck hepatitis virus posttranscriptional regulatory element sense (SEQ ID NO: 47) for A + B backbones, or the reverse complement (SEQ ID NO: 17) for C and D backbones. The Constructs A and B allow transgene expression in producer cells while constructs C and D have no transgene expression in these cells. AUG = start codon, CAU = reverse complement of start codon, pDNA = plasmid DNA, and ssRNA = single stranded RNA.

[0050] Figure 2 shows that the comparative example CCL. LP1. coABCB4 construct hinders the production of the lentiviral vectors needed for PFIC3. (A) The cloning of coABCB4 into the CCL. LP1 vector was confirmed by digestion with BstBT, BspTI and EcoRI and (B) Sanger sequencing. (C) Lentiviral vectors were produced by transient transfection of HEK293T cells using the CCL. LP1. coABCB4 transgene plasmid and Verapamil inhibition. The vectors were harvested 48 hours post-transfection and tittered. No MDR3 inhibitor hinders vector production, while inhibition at various verapamil concentrations increased the vector yield to just above 1x104TU / ml.

[0051] Figure 3 shows that the example CCL. LPa. coABCB4 construct, comprising the novel LPa promoter, improves vector titres and MDR3 localisation at bile canaliculi in HepG2 ABCB4~ / ~ cells. Sanger sequencing was used to confirm the successful cloning of (A) LPa promoter (SEQ ID NO: 3) and (B) LPE promoter (SEQ ID NO: 45) containing the novel LPa promoter sequence and an enhancer region. (C) Lentiviral vectors were produced by transient transfection of Hek293T cells using the example CCL. LP1. coABCB4, CCL.hAAT. coABCB4, CCL. LPa. coABCB4 or CCL. LPE. COABCB4 as transgene plasmids along with Verapamil inhibition. The vectors were harvested 48 hours post-transfection, concentrated and titered. Titration has revealed that the CCL. LPa. coABCB4 or CCL. LPE. COABCB4 vectors have increased yields over 1.5x107TU / ml. Transduction of HepG2 ABCB4~ / ~ with the produced vectors at a multiplicity of infection of 1 had shown that 48 hours post transduction cells had on average similar vector copy number (D), but treatments with the example CCL. LPa. coABCB4 or CCL. LPE. COABCB4 vectors have displayed lower levels of ABCB4 expression (E-F). (G) Immunofluorescence analysis images taken at 25x magnification showing localisation of MDR3.

[0052] Figure 4 shows the in vivo analysis of the example CCL. LPa. Transgene vectors comprising the novel promoter of the invention. Three weeks old FVB.129P2-Abcb4fmie°7J were injectedwith 1x1010TU / kg CCL. LPa. Luc. GFP (LPa) and received intraperitoneal Luciferin injections 2 days thereafter along wild type animals. (A) Live I VIS imaging and (B) organ I VIS imaging where black areas representing low expression levels and white areas in the middle of dark regions representing high levels of expression. Neonate FVB. 29P2-Abcb4tm1Bor / J mice were also injected with CCL. LPa. coABCB4 (LV, SEQ ID NO: 29) or control CCL. LPa. Luc. GFP (LG, SEQ ID NO: 31) and monitored for 18 weeks along wild type mice. (C) Alanine aminotransferase levels at 6 weeks and at 18 weeks (harvest). (D) Vector copy number and (E) ABCB4 gene expression measured in the livers of experimental animals.

[0053] Figure 5 shows that example lentiviral vectors for PFIC3, containing a reverse transgene expression cassette, show adequate titres and ability for transgene expression in HepG2 cells. Sanger sequencing cloning confirmation of (A).miR-G-LPaH (SEQ ID NO: 32) (B) miR-coA4-LPaH (SEQ ID NO: 33 - coABCB4 Version 3 (C) miR-wA4-LPa (SEQ ID NO: 36) (D) miR-wtA4-LPaH (SEQ ID NO: 34 - wtABCB4 Version 3) (E) miR-GFP. Luc-LPaH (SEQ ID NO: 39).

[0054] (F) Lentiviral vectors were produced by transient transfection of Hek293T cells using the miR-G-LPaH (G. LPaH), miR-GFP. Luc-LPaH (GL. LPaH), miR-coA4-LPaH (coA4. LPaH), miR-coA4-LPa (coA4. LPa), miR-wA4-LPaH (wA4. LPaH) and miR-wA4-LPa (wA4. LPa) as transgene plasmids. The example vectors were harvested 48 hours post-transfection, concentrated and tittered. Titration has revealed that the version 3 backbones carrying the wild type ABCB4 gene (SEQ ID NO: 8) generated titres of above 1x109TU / ml. (G) Immunofluorescence imaging of HepG2 cells with or without the miR-G-LPaH transduction.

[0055] Figure 6 shows that the example miR-wtA4-LPaH vectors rescue the PFIC3 phenotype of in vivo. (A) Neonate FVB. '\29P2-Abcb4tm1Bor / J pre-treated with 0.12g / kg intraperitoneal injections of clodronate liposomes were injected at P3-4 with 3x101oTU / kg of miR-wtA4-LPaH (LV, n=6) and placed at 4 weeks for 9 days on a 0.10% CA diet to exacerbate the cholestatic phenotype along with WT (n=5) and untreated KO (n=6) animals. (B) Animal survival from week 4. The treated and WT animals survived till the end of the experiment at 12 weeks. (C) Weight change over the first CA administration for WT (n=5), KO (n=3) and LV (n=2). (D) Alanine aminotransferase and (E) alkaline phosphatase levels in serum collected from all experimental animals at 5-6 weeks. (F) Representative image of plasma opacity in KO and LV treated animals at harvest. The darker KO plasma contains high levels of the bilirubin pigment as confirmed by (G) direct bilirubin levels measured in the serum at harvest. (H) Vector copy number in the livers of treated animals. (I) Picrosirius red fibrosis staining of animal livers. Dark areas represent fibrotic regions quantified in (J) by measuring the percentage of liver fibrosis areas. Graphs (D-H) display unique values, medians and IQRs.Figure 7 shows the example miR-wtA4-LPaH vectors rescue the excretion of phosphatidyl choline in the bile and the expression of the ABCB4 protein in the liver of PFIC3 mice. Neonate FVB.129P2-Abcb4fm76o7J pre-treated with 0.12g / kg intraperitoneal injections of clodronate liposomes were injected at P3-4 with 3x1010TU / kg of miR-wtA4-LPaH (LV) and placed at 4 weeks for 9 days on a 0.10% CA diet to exacerbate the cholestatic phenotype along with WT and untreated KO animals. (A) Phosphatidyl choline levels measured by mass spectrometry in bile samples collected at harvest from WT (n=3), KO (n=4) and LV (n=4) mice. (B) ABCB4 protein level measured by mass spectrometry in liver samples collected at harvest from randomly selected mice (n=3) per group.

[0056] Figure 8 shows safety results following integration site analysis experiments performed on primary human hepatocytes after treatment with the miR-wtA4-LPaH (SEQ ID NO: 34). Prior to analysis the cells were treated with the miR-wtA4-LPaH (SEQ ID NO: 34) at MOI 2 and MOI 20. (A) Shannon diversity score measuring integration site variety. (B) Integration site frequency with black close to x-axis representing sum of integration events with a frequency below 1%. (C) Total integration sites measured in samples. Circle packing plots show the genes with Gene Relative Abundance (GRA) greater than 0.0001 in (D) cells treated at MOI 2 and (E). (F) Tables showing genes with top 5 integration frequencies in each replicate.

[0057] Figure 9 shows that the CCL. LP1.coABCB11 lentiviral vector has a significantly lower titre compared to the CCL. LP1. GFP vector, likely due to the expression of ABCB11 protein in the Hek293T cells during vector production. (A) Unconcentrated lentiviral vector titre obtained from production in Hek293T cells using the CCL. LP1. GFP (GEW, SEQ ID NO: 74) and the CCL. LP1. coABCBH (ABCB11, SEQ ID NO: 69). (B) Western blot showing that the CCL. LP1.coABCB77 (ABCB11, SEQ ID NO: 69) expresses ABCB11 in producer cells in comparison to the miR-coABCB11-LPaH (SEQ ID NO: 70).

[0058] Detailed Description

[0059] Although any materials, materials for use, and methods similar or equivalent to those described herein can be used in the practice or testing of the present disclosure, the preferred embodiments, materials and methods are now described. Other features, objects and advantages of the disclosure will be apparent from the description. In the description, the singular forms also include the plural unless the context clearly dictates otherwise. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs unless stated otherwise.The term “vector” refers to any vehicle that can be used to deliver the nucleic acid material of interest into cells.

[0060] A lentiviral vector sequence defined herein relates to the genome of the lentiviral vector that can be used as a gene therapy.

[0061] A lentiviral vector disclosed herein relates to a lentivirus vector comprising the lentiviral vector sequence disclosed herein. This can be used as a gene therapy.

[0062] For any sequence described herein, the complementary sequence or reverse complement is also implicitly disclosed and considered part of the disclosure.

[0063] The terms "treatment" and "treating" herein refer to an approach for obtaining beneficial or desired results in a subject, which includes a prophylactic benefit and a therapeutic benefit.

[0064] “Therapeutic benefit” refers to eradication, amelioration or slowing the progression of the underlying disorder being treated. Also, a therapeutic benefit is achieved with the eradication or amelioration of one or more of the physiological symptoms associated with the underlying disorder such that an improvement is observed in the subject, notwithstanding that the patient may still be afflicted with the underlying disorder.

[0065] “Prophylactic benefit” refers to delaying or eliminating the appearance of a disease or condition, delaying or eliminating the onset of symptoms of a disease or condition, slowing, halting, or reversing the progression of a disease or condition, or any combination thereof. In the context of the present invention, the prophylactic benefit or effect may involve the prevention of the condition or disease. The vector or pharmaceutical composition described herein may be administered to a subject at risk of developing a disease described herein, or to a subject reporting one or more of the physiological symptoms of a disease described herein, even though a diagnosis of this disease may not have been made.

[0066] The term "effective amount" or "therapeutically effective amount" refers to the amount of the vector or pharmaceutical composition needed to bring about an acceptable outcome of the therapy as determined by reducing the likelihood of the disease as measurable by clinical, biochemical or other indicators that are familiar to those trained in the art. The therapeutically effective amount may vary depending upon the condition, the severity of the condition, thesubject, e.g., the weight and age of the subject and the mode of administration and the like, which can readily be determined by one of ordinary skill in the art.

[0067] The term "subject" refers to any suitable subject, including any animal, such as a mammal. In preferred embodiments described herein, the subject is a human. In some embodiments, the subject may be a test subject, e.g., selected from mouse, a rat, a canine, a pig, a non-human primate or a guinea pig.

[0068] The term "comprising" (and related terms such as "comprise" or "comprises" or "having" or "including") includes those embodiments, for example, an embodiment of any composition of matter, composition, method, or process, or the like, that "consist of” or "consist essentially of” the described features. The term “comprises” or “comprising” can be used interchangeably with “includes”.

[0069] Mutated splice sites herein refers to any genetic mutation / alteration (e.g., from a naturally occurring DNA sequence) that occurs at the boundary of an exon and an intron. For example, the mutation of the donor splice site is such that the 5’ end of the intron is not GT and mutation of the acceptor splice site is such that the 3’ end of the intron is not AG. Mutation of the splice site is such that the splice site is no spliced, or recognised by splicing machinery, in the cell.

[0070] The term “identical” and “identity” refers to the overall relatedness between polymeric molecules, e.g., between polynucleotide molecules (e.g., DNA molecules and / or RNA molecules) and / or between polypeptide molecules. Calculation of the percent identity of two polynucleotide sequences, for example, can be performed by aligning the two sequences for optimal comparison purposes (e.g., gaps can be introduced in one or both of a first and a second nucleic acid sequences for optimal alignment and non-identical sequences can be disregarded for comparison purposes). The nucleotides at corresponding nucleotide positions are then compared. When a position in the first sequence is occupied by the same nucleotide as the corresponding position in the second sequence, then the molecules are identical at that position. The percent identity between the two sequences is a function of the number of identical positions shared by the sequences, taking into account the number of gaps, and the length of each gap, which needs to be introduced for optimal alignment of the two sequences. The comparison of sequences and determination of percent identity between two sequences can be accomplished using a mathematical algorithm. For example, in a preferred embodiment, the percent identity between two nucleotide sequences can be determined using an alignment algorithm, e.g., such as the BLAST sequence analysis tool.The term “encode” refers broadly to any process whereby the information in a polymeric vector is used to direct the production of a second molecule that is different from the first. In the context of this invention, a lentiviral vector that encodes for a protein refers to a lentiviral vector comprising a gene (which can also be referred to as a transgene) which specifies the sequence of amino acids for the protein of interest.

[0071] vg / ml defined herein refers to vector genome copies per millilitre. TU / ml refers to transducing units per ml.

[0072] Promoter

[0073] In a first aspect of the present invention, there is provided a promoter sequence comprising at least a portion of a ABCB4 promoter or reverse complement thereof, wherein the splice sites of the ABCB4 promoter have been mutated. In some embodiments, the ABCB4 promoter is a human ABCB4 promoter sequence.

[0074] Since the splice sites have been mutated, the promoter sequence does not comprise a sequence which is identical to a portion of the naturally occurring ABCB4 promoter sequence or reverse complement thereof, i.e., SEQ ID NO 23 orSEQ ID NO: 24. The promoter sequence may instead comprise a sequence corresponding to SEQ ID NO: 23 or SEQ ID NO: 24 but wherein the splice donor and splice acceptor sites have been mutated. In an example, both the splice donor and splice acceptor sites of SEQ ID NO: 23 have been mutated, to CO.

[0075] In some embodiments, at least a portion of theABCB4 promoter, or the entireABCB4 promoter, comprises or has at least 95 % sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity to SEQ I D NO: 1 or 2. This corresponds to the LPa promoter and reverse LPa promoter sequence defined herein, where the splice sites have been mutated. SEQ ID NO: 2 is the reverse complement of SEQ ID NO:1. SEQ ID NO: 2 can be utilised in lentiviral vector sequences where the promoter sequence and the transgene are in reverse orientation to the direction of transcription of the lentiviral vector sequence (i.e., according to the third aspect). SEQ ID NO: 1 can be utilised in lentiviral vector sequences where the promoter sequence and the transgene are in the same orientation to the direction of the transcription of the lentiviral vector sequence (i.e., lentiviral vector genome (i.e., according to the second aspect)

[0076] In some embodiments, the promoter sequence comprises or has a sequence corresponding to SEQ ID NO: 1 with the proviso that the sequence is not SEQ ID NO: 23. This is the LPapromoter defined herein wherein the donor splice site is not GT and the acceptor splice site is not AG. In some embodiments, the LPa promoter has a sequence corresponding to SEQ ID NO: 3, wherein the donor splice site and the acceptor splice site has been mutated to CC.

[0077] In some embodiments, the promoter sequence or has a sequence corresponding to SEQ ID NO: 2 with the proviso that the sequence is not SEQ I D NO: 24. This is the reverse complement of the LPa promoter defined herein. In some embodiments, the promoter sequence comprises or has a sequence corresponding to SEQ ID NO: 4 which is the reverse complement of SEQ ID NO: 3.

[0078] In some embodiments, at least a portion of theABCB4 promoter, or the entireABCB4 promoter, comprises or has at least 95 % sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity to SEQ ID NO: 3 or SEQ ID NO: 4. As indicated above, SEQ ID NO: 4 is the reverse complement of SEQ ID NO:3. SEQ ID NO: 4 can be utilised in lentiviral vectors where the promoter sequence and the transgene are in reverse orientation to the direction of transcription of the lentiviral vector sequence (i.e., according to the third aspect). SEQ ID NO: 3 can be utilised in lentiviral vectors where the promoter sequence and the transgene are in the same orientation to the direction of the transcription of the lentiviral vector sequence (i.e., lentiviral vector genome). This is an example LPa promoter sequence, where the splice sites have been mutated to CC and GG respectively.

[0079] In some embodiments, the splice site or the promoter is mutated such that the splice donor site is not GT. For example, in SEQ ID NO: 1, SEQ ID NO:3 or variants thereof, the splice donor site at sequence positions 372-373 is not GT. In some embodiments, the splice donor site at sequence positions 372-373 is selected from AA, AT, AG, AC, CA, CT, CG, CC, TA, TT, TG, TC, GA, GC or GG, (e.g., CC in SEQ ID NO: 3).

[0080] If a reverse complement of the ABCB4 promoter is used (in lentiviral vectors where the promoter sequence and the transgene are in reverse orientation to the direction of transcription of the lentiviral vector sequence), the splice site is mutated such that the reverse complement of the splice donor site is not AC. For example, in SEQ ID NO: 2, SEQ ID NO:4 or variants thereof, the reverse complement of the splice donor site at positions 167-168 is not AC. In some embodiments, the reverse complement of the splice donor site at positions 167-168 is selected from GG, GA, GC, GT, AA, AT, AG, TG, TA, TC, TT, CA, CC, CG, CT (e.g., GG in SEQ ID NO: 4).In some embodiments, the splice site or the promoter is mutated such that the splice acceptor site is not AG. For example, in SEQ ID NO: 1, SEQ ID NO:3 or variants thereof, the splice acceptor site at positions 532-533 is not AG. In some embodiments, the splice acceptor site at positions 532-533 is not AG and is selected from AA, AT, AC, CA, CT, CG, CC, TA, TT, TC, GA or GT (e.g., CC in SEQ ID NO: 3).

[0081] If a reverse complement of the ABCB4 promoter is used (in lentiviral vectors where the promoter sequence and the transgene are in reverse orientation to the direction of transcription of the lentiviral vector sequence), the splice site is mutated such that the reverse complement of the splice donor site is not GT. For example, in SEQ ID NO: 2, SEQ ID NO: 4 or variants thereof, the reverse complement of the splice acceptor site at positions 7-8 is not GT. In some embodiments, in SEQ ID NO: 2, SEQ ID NO: 4 or variants thereof, the reverse complement of the splice acceptor site at positions 7-8 is not GT and is instead selected from CC, TC, GC, AC, TT, TA, CT, CA, AT, GA, AA, TG, GG, CG, or AG, (e.g., GG in SEQ ID NO: 4).

[0082] In some embodiments, the promoter sequence further comprises an enhancer sequence. In some embodiments, the enhancer sequence is a HCR enhancer sequence. In some embodiments, the HCR enhancer sequence is defined by SEQ ID NO: 5 or a reverse complement thereof. In some embodiments, the HCR enhancer sequence has at least 95% sequence identity, or at least 96% identity, or at least 97% identity, or at least 98% identity, or at least 99% identity, or at least 100% sequence identity with SEQ ID NO:

[0083] 5 or the reverse complement thereof.

[0084] In some embodiments, the promoter sequence comprises the LPa promoter (e.g., SEQ ID NO: 1 or 3) and the HCR enhancer sequence (SEQ ID NO: 5). In some embodiments, the promoter comprises or has a sequence with at least 95% sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity, or 100% sequence identity to SEQ ID NO: 6.

[0085] In some embodiments, the promoter sequence comprises the reverse complement of the LPa promoter (e.g., SEQ ID NO: 2 or 4) and the reverse complement of the HCR enhancer sequence (reverse complement of SEQ ID NO: 5). In some embodiments, the promoter comprises or has a sequence with at least 95% sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity, or 100% sequence identity to SEQ ID NO: 7.In some embodiments, the promoter comprises an enhancer sequence which corresponds to an enhancer sequence found 129bp upstream of the natural ABCB4 promoter, i.e., corresponding to SEQ ID NO: 66. In such embodiments, the promoter sequence has a sequence that has at least 95% sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity, or 100% sequence identity, with SEQ ID NO: 45 or a reverse complement thereof. The combination of the LPa promoter + enhancer sequence from the natural ABCB4 promoter is referred to as the LPe promoter herein.

[0086] Lentiviral Vector Sequence and Lentiviral Vector

[0087] Disclosed herein is a lentiviral vector sequence comprising the promoter sequence defined herein (i.e., according to the first aspect). The lentiviral vector sequence is the sequence used in a lentiviral vector.

[0088] In a second aspect of the present invention, there is provided a lentiviral vector sequence comprising a promoter sequence defined herein (i.e., of the first aspect) which is operably linked with a transgene. The promoter sequences comprises at least a portion of a ABCB4 promoter, wherein the splice sites have been mutated as disclosed herein. The promoter sequence may be or comprise the LPa promoter defined herein.

[0089] In a third aspect of the present invention, there is provided a lentiviral vector sequence comprising a promoter sequence of the first aspect, comprising the promoter sequence defined herein (i.e., of the first aspect) which is operably linked with a transgene, wherein the promoter sequence and the transgene are in reverse orientation to the direction of transcription of the lentiviral vector (i.e., lentiviral vector genome). This may otherwise be described as a lentiviral vector sequence having a reverse transgene expression cassette. The promoter sequence used is the reverse complement of at least a portion of a ABCB4 promoter, wherein the splice sites have been mutated. The promoter sequence may correspond to, or comprise, the reverse complement of the LPa promoter defined herein.

[0090] Enhancer

[0091] In some embodiments, the lentiviral vector sequence further comprises an enhancer sequence. In some embodiments, the enhancer sequence is an HCR enhancer sequence. In some embodiments, the HCR enhancer sequence is defined by SEQ ID NO: 5 or a reverse complement thereof. In some embodiments, the HCR enhancer sequence has at least 95%sequence identity, or at least 96% identity, or at least 97% identity, or at least 98% sequence identity, or at least 99% sequence identity, or at least 100% sequence identity with SEQ ID NO: 5 or the reverse complement thereof.

[0092] In some embodiments, the lentiviral vector sequence comprises the LPa promoter (e.g., SEQ ID NO 1 or 3) and the HCR enhancer sequence, or the reverse complement of the LPa promoter (e.g., SEQ ID NO 2 or 4) and the reverse complement of the HCR enhancer sequence. In some embodiments, the lentiviral vector sequence comprises a sequence with at least 95% sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity, or 100% sequence identity to SEQ ID NO: 6 or a reverse complement thereof (SEQ ID NO: 7).

[0093] In some embodiments, the lentiviral vector comprises an enhancer sequence which corresponds to an enhancer sequence found 129bp upstream of the natural ABCB4 promoter, i.e., corresponding to SEQ ID NO: 66. In such embodiments, the lentiviral vector has a sequence that has at least 95% sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity, or 100% sequence identity, with SEQ ID NO: 45 or a reverse complement thereof.

[0094] Transgene

[0095] In some embodiments, the transgene is a therapeutic protein (i.e., a protein used to replace a protein that is abnormal or deficient in a particular disease).

[0096] In some embodiments, the transgene encodes for a liver protein and / or a protein expressed by a hepatocyte cell.

[0097] In some embodiments, the transgene encodes for a liver transmembrane protein. In some embodiments, the transgene encodes for an ATP binding cassette transporter. An ATP binding cassette transporter may otherwise be referred to as an ABC transporter in the art. The ATP binding cassette transporter protein may be of the ABCA subfamily, ABCB subfamily, ABCC subfamily, ABCD subfamily, ABCE / ABCF subfamily or ABCG subfamily. In some embodiments, the ATP binding cassette transporter is of the ABCB subfamily

[0098] In some embodiments, the ATP binding cassette transporter is ABCB4, ABCB11 or ATP8B1, ABCC2, ABCD3, ABCG5, ABCG8, ABCB6, ABCC9, ABCA1 or ABCA12. In some embodiments, the ATP binding cassette transporter is ABCB4, ABCB11 or ATP8B1. In someembodiments, the ATP binding cassette transporter is ABCB4 or ABCB11. In some embodiments, the ATP binding cassette transporter is ABCB4.

[0099] In some embodiments, the transgene encodes for human ABCB4, i.e., human wild-type ABCB4. In some embodiments, the transgene comprises or has a sequence with at least 95% sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity, or 100% sequence identity, to SEQ ID NO: 8. SEQ ID NO: 8 is the reverse complement of wtABCB4 and is used where the promoter sequence and the transgene are in the reverse orientation to the direction of transcription of the lentiviral vector sequence (i.e., lentiviral vector genome)

[0100] . In some embodiments, the transgene comprises or has a sequence with at least 95% sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity, or 100% sequence identity with SEQ ID NO: 43. SEQ ID NO: 43 is the wtABCB4 sequence and may be used where the promoter sequence and the transgene are in the same orientation to the direction of transcription of the lentiviral vector sequence (i.e., lentiviral vector genome).

[0101] In some embodiments, the transgene encodes for a codon-optimised ABCB4. In some embodiments, the transgene comprises or has a sequence with at least 95% sequence identity, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 9 or the reverse complement thereof. SEQ ID NO: 9 is the reverse complement of codon-optimised ABCB4 and is used where the promoter sequence and the transgene are in the reverse orientation to the direction of transcription of the lentiviral vector sequence (i.e., lentiviral vector genome)

[0102] In some embodiments, the transgene coding sequence has at least 95% sequence identity, or, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 44. SEQ ID NO: 44 is a codon-optimised ABCB4 sequence and may be used where the promoter sequence and the transgene are in the same orientation to the direction of transcription of the lentiviral vector sequence (i.e., lentiviral vector genome).

[0103] In some embodiments, the transgene encodes for human ABCB11, i.e., human wild-type ABCB11 or has a sequence with at least 95% sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity, or 100% sequence identity, to a human wild-type ABCB11.In some embodiments, the transgene encodes for a codon-optimised ABCB11. In some embodiments, the transgene comprises or has a sequence with at least 95% sequence identity, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or 100% sequence identity to SEQ ID NO: 72 or SEQ ID NO: 73. SEQ ID NO: 72 is a codon-optimised ABCB11. SEQ ID NO: 73 is the reverse complement of a codon-optimised ABCB11.

[0104] In some embodiments, the transgene encodes for a diagnostic protein or marker protein. In some examples, the transgene encodes for a fluorescent protein, e.g., GFP.

[0105] miR-binding sites

[0106] In some embodiments, the lentiviral vector sequences defined herein further comprise one or more miRNA binding sequences, or one or more repeat miRNA binding sequences, or the reverse complement thereof. In some embodiments, the miRNA binding sequence is selected from miR-142, miR-126-3pT or a combination thereof. In some embodiments, the lentiviral vector sequence comprises two or more, or three or more, or four or more of the miRNA binding sequence(s), e.g., at least two of miR-142, and / or at least two of miR-126-3pT. In some embodiments, the vector comprises four miR-142 binding sequences and / or four miR-126-3pT binding sequences. miRNA binding sequences may be used interchangeably with miRNA sequences herein.

[0107] In some embodiments, the lentiviral vector sequence comprises a sequence with at least 95% sequence identity, or at least 96% sequence identity, or at least 97% sequence identity, or at least 98% sequence identity, or at least 99% sequence identity, or 100% sequence identity to SEQ ID NO: 10-14, ora reverse complement thereof.

[0108] Post-transcriptional regulatory element

[0109] In some embodiments, the lentiviral vector sequence defined herein further comprises a post-transcriptional regulatory element, or reverse complement thereof. Any suitable post-transcriptional regulatory element may be used. In some embodiments the post-transcriptional regulatory element is the woodchuck hepatitis virus post-transcriptional regulatory element. In some embodiments, the lentiviral vector sequence comprises a sequence corresponding to SEQ ID NO 47, or the reverse complement thereof (SEQ ID NO: 17). The reverse complement will be used. The reverse complement of the post-transcriptional regulatory element (e.g., SEQ ID NO: 17) is used where the promoter sequence and the transgene are in the reverse orientation to the direction of transcription of the lentiviral vector sequence (i.e., lentiviral vector genome).

[0110] Poly A signalIn some embodiments, the lentiviral vectors defined herein further comprise a polyA signal. Any suitable polyA signal may be used. In some embodiments, the lentiviral vector sequence defined herein comprise the bovine growth hormone polyadenylation signal (bGH)poly(A) signal or the reverse complement thereof. In some embodiments, the lentiviral vector comprises SEQ ID NO: 21. This is the reverse complement of the bGH(poly)(A) signal.

[0111] Type of lentiviral vector, LTR sequences and packaging sequences

[0112] In some embodiments, the lentiviral vector sequence and lentiviral vector is based on human immunodeficiency virus (HIV), in other words, it is a HIV-derived vector sequence. The lentiviral vector sequence may be based on HIV of type 1 or type 2, preferably HIV-1.

[0113] Any other suitable lentiviral vector sequences or lentiviral vector may be used. In some embodiments, the lentiviral vector sequence or lentiviral vector is based on simian immunodeficiency virus (SIV), feline immunodeficiency virus (FIV), equine infectious anaemia virus (EIAV), caprine arthritis encephalitis virus (CAEV), bovine immunodeficiency virus (BIV), or jembrana disease virus (JDV).

[0114] The human immunodeficiency virus (HlV)-based vector sequence typically comprises a truncated 5’ long terminal repeat (LTR) from HIV. In some embodiments, the lentiviral vector sequence comprises SEQ ID NO: 18.

[0115] The human immunodeficiency virus (HlV)-based vector sequence typically comprises a truncated 3’ long terminal repeat (LTR) from HIV. In some embodiments, the lentiviral vector sequence comprises SEQ ID NO: 22.

[0116] The human immunodeficiency virus (HlV)-based vector sequence typically comprises the HIV-1 packaging signal of HIV-1. In some embodiments, the lentiviral vector sequence comprises SEQ ID NO: 19.

[0117] The human immunodeficiency virus (HlV)-based vector sequence typically comprises the Rev response element (RRE) from HIV-1. In some embodiments, the lentiviral vector sequence comprises SEQ ID NO: 46.

[0118] The human immunodeficiency virus (HlV)-based vector sequence typically comprises the central polypurine tract (cPPT) and the central termination sequence (CTS) of HIV-1. In some embodiments, the lentiviral vector sequence comprises SEQ ID NO: 20.In some embodiments, the LTR-LTR sequence of the lentiviral vector sequence has a sequence which is at least 95%, or at least 96%, or at least 97%, or at least 98%, or at least 99%, or is 100% identical with SEQ ID NO: 15 or SEQ ID NO: 16 or SEQ ID NO: 71.

[0119] Disclosed herein is a lentiviral vector comprising the lentiviral vector sequence defined herein.

[0120] Exemplary Version 3 backbones

[0121] “Version 3” backbones herein are preferred lentivirus vector sequences of the invention in accordance with the third aspect, and comprise a reverse expression cassette.

[0122] In some embodiments, a lentiviral vector sequence comprises a sequence comprising at least, in the direction

[0123]

[0124] 5’ 3’:

[0125] A reverse transgene sequence (such as the ABCB4 transgene, e.g., SEQ I D NO: 8 or 9, or the ABCB11 transgene, e.g. SEQ ID NO: 73), and

[0126] The reverse complement of the LPa promoter, e.g., SEQ ID NO: 2 or 4, and

[0127] optionally the reverse complement of an enhancer, such as an HCR enhancer, e.g., the reverse complement of SEQ ID NO: 5.

[0128] In some embodiments, a lentiviral vector sequence comprises a sequence comprising at least in the direction

[0129]

[0130] 5’ 3’:

[0131] the reverse complement of a post transcriptional regulatory element, such as WPRE, e.g., SEQ ID NO: 17

[0132] a reverse transgene sequence (such as a ABCB4 transgene, e.g., SEQ ID NO: 8 or 9, or the ABCB11 transgene, e.g. SEQ ID NO: 73),

[0133] the reverse complement of the LPa promoter, e.g., SEQ ID NO: 2 or 4, and

[0134] optionally the reverse complement of an enhancer, such as an HCR enhancer, e.g., the reverse complement of SEQ ID NO: 5.

[0135] In some embodiments, a lentiviral vector sequence comprises a sequence comprising at least: the reverse complement of a poly A signal, such as the bGH poly(A) signal (e.g., SEQ ID NO: 21)

[0136] the reverse complement of a post transcriptional regulatory element, such as WPRE, e.g., SEQ ID NO: 17

[0137] a reverse transgene sequence (such as a ABCB4 transgene, e.g., SEQ ID NO: 8 or 9, or the ABCB11 transgene, e.g. SEQ ID NO: 73),the reverse complement of the LPa promoter, e.g., SEQ ID NO: 2 or 4, and

[0138] optionally the reverse complement of an enhancer, such as an HCR enhancer, e.g., the reverse complement of SEQ ID NO: 5.

[0139] In some embodiments, a lentiviral vector sequence comprises a sequence comprising at least in the direction

[0140]

[0141] 5’ 3’:

[0142] the reverse complement of a poly A signal, such as the bGH poly(A) signal (e.g., SEQ ID NO: 21)

[0143] a repeat miRNA binding sequence containing one or more miRNA binding sites, e.g., SEQ ID NO: 14

[0144] the reverse complement of a post transcriptional regulatory element, such as WPRE, e.g., SEQ ID NO: 17

[0145] a reverse transgene sequence (such as a ABCB4 transgene, e.g., SEQ ID NO: 8 or 9, or the ABCB11 transgene, e.g. SEQ ID NO: 73),

[0146] the reverse complement of the LPa promoter, e.g., SEQ ID NO: 2 or 4, and

[0147] optionally the reverse complement of an enhancer, such as an HCR enhancer, e.g., the reverse complement of SEQ ID NO: 5.

[0148] In some embodiments, the lentiviral vector sequence comprises:

[0149] A 5’-LTR sequence, e.g., SEQ ID NO: 18

[0150] A packaging signal, e.g., SEQ ID NO: 19

[0151] A central polypurine tract and central termination sequence, e.g., SEQ ID NO: 20

[0152] An RRE, e.g., SEQ ID NO: 46

[0153] the reverse complement of a poly A signal, such as the bGH poly(A) signal (e.g., SEQ ID NO: 21)

[0154] a repeat miRNA binding sequence containing one or more miRNA binding sites, e.g., SEQ ID NO: 14

[0155] the reverse complement of a post transcriptional regulatory element, such as WPRE, e.g., SEQ ID NO: 17

[0156] a reverse transgene sequence (such as a ABCB4 transgene, e.g., SEQ ID NO: 8 or 9, or the ABCB11 transgene, e.g. SEQ ID NO: 73),

[0157] the reverse complement of the LPa promoter, e.g., SEQ ID NO: 2 or 4,

[0158] optionally the reverse complement of an enhancer, such as an HCR enhancer, e.g., the reverse complement of SEQ ID NO: 5, and

[0159] a 3’-LTR sequence, e.g., SEQ ID NO: 22

[0160] Medical Use or Methods of TreatmentThe lenti viral vectors sequence of the present disclosure (i.e., according to the second or third aspect) or the lentiviral vectors of the fourth aspect, can be for use in therapy and / or may be used in a method of treatment.

[0161] The lentiviral vector sequence of the present disclosure (i.e., according to the second or third aspect), or the or the lentiviral vectors of the fourth aspect, may also be for use in the treatment of a disease. Also disclosed herein are methods of treatment for treating a disease, comprising administering to a subject a pharmaceutically effective amount of the lentiviral vector sequences disclosed herein (i.e., of the second or third aspect) or the lentiviral vectors disclosed herein (i.e., of the fourth aspect).

[0162] In some embodiments, the disease is a liver disease.

[0163] In some embodiments, the disease is cholestasis. The cause of cholestasis may be selected from familial intrahepatic cholestasis, pregnancy, cancer, liver disease, biliary trauma, gallstones, biliary dyskinesia, acute hepatitis, cystic fibrosis, drug-induced cholestasis or secondary syphilis. In some embodiments, the disease is Dubin-Johnson syndrome. In some embodiments, the disease is congenital bile acid synthesis defect 5. In some embodiments, the disease is sitosterolaemia.

[0164] In some embodiments, the drug induced cholestasis is induced by paracetamol / acetaminophen, antifungals or antibiotics (e.g., penicillin, sulfonamides), antidiabetic drugs, anti-inflammatory drugs, cardiovascular drugs (e.g., statins), psychotropic drugs (e.g., chlorpromazine), birth control, hormones (e.g., anabolic steroids or estrogen), cancer.

[0165] In some embodiments, the disease is familial intrahepatic cholestasis (FIH), particularly progressive familial intrahepatic cholestasis (PFIC). In some embodiments, the progressive familial intrahepatic cholestasis is PFIC1, PFIC2, PFIC3 or PFIC4. In some embodiments, the transgene encodes for ABCB4 and the progressive familial intrahepatic cholestasis is PFIC3. In some embodiments, the transgene encodes for ABCB11 and the progressive familial intrahepatic cholestasis is PFIC2. In some embodiments, the transgene encodes for APT8B1 and the progressive familial intrahepatic cholestasis is PFIC1.

[0166] In some embodiments, the transgene encodes for ABCC2, and the disease to be treated is Dubin-Johnson syndrome. In some embodiments, the transgene encodes for ABCD3, and thedisease to be treated is Congenital bile acid synthesis defect 4. In some embodiments, the transgene encodes for ABCG5 or ABCG8 and the disease to be treated is Sitosterolaemia.

[0167] In some embodiments, the disease is a disease associated with a mutated ABC protein and / or aberrant expression of a mutated ABC protein. This may otherwise be referred to as an ABC transporter disorder.

[0168] In some embodiments, the ABC protein is ABCB4, and the disease associated with mutated ABCB4 and / or aberrant expression of ABCB4 is PFIC3, ICP3 (intrahepatic cholestasis of pregnancy type 3), or GBD1 (gallbladder disease 1) or low phospholipid associated cholelithiasis, preferably PFIC3.

[0169] In some embodiments, the ABC protein is ABCB11, and the disease associated with mutated ABCB11 and / or aberrant expression of ABCB11 is PFIC2 or BRIC2 (benign recurrent intrahepatic cholestasis)

[0170] Host cell and sequence

[0171] Also disclosed herein is a cell or host cell comprising the lentiviral vector sequence of the second and third aspect, or the lentivirus vector of the fourth aspect. The cell may be a liver cell, e.g., a hepatocyte cell. The cell may be a producer cell. The cell may be in vivo or in vitro.

[0172] Use of the vectors

[0173] Also defined herein is the use of the promoter sequence as defined in the first aspect, or the lentiviral vector sequence as defined in the second or third aspect, to improve lentiviral titre. The lentiviral titre may be improved as compared to a lentiviral titre where the known LP1 promoter is used. In some embodiments, use of the promoter sequence or lentiviral vector sequence as defined herein results in a titre of at least 1 x 108TU / ml, or at least 5 x 108TU / ml, or at least 1 x 109TU / ml.Examples

[0174] The present inventors have developed a novel promoter and novel lentiviral vector sequence to produce a novel lentiviral vector. The novel promoter and novel lentiviral vector sequence can be used in lentiviral gene therapies. Below, we describe methods, rationale and experiments used in support of an example lentiviral vectors expressing the ABCB4 gene, which can be used as a lentiviral gene therapy for PFIC3.

[0175] Results

[0176] Comparative Example 1 - Lentiviral Vector - CCL. LP1.coABCB4

[0177] A CCL. LP1,coABCB4 lentiviral vector backbone comprising the known promoter LP1 impedes vector production

[0178] We aimed to develop and test a liver directed lentiviral gene therapy for treating PFIC3. For this, a self-inactivating lentiviral vector, named CCL. LP1. coABCB4 (SEQ ID NO: 28 -CCL. LP1,coABCB4_v1, Figure 1), carrying the codon optimised ABCB4 (SEQ ID NO: 9) under the control of a known liver-specific Liver Promoter 1 (LP1, SEQ ID NO: 42) (18,19) was first generated. The LP1 promoter (SEQ ID NO: 42) composed of the human apolipoprotein hepatic control region (HCR, SEQ ID NO: 41) and the human alpha-1-antitrypsin (hAAT) promoter (SEQ ID NO: 40) (20) was chosen for initial experiments as it has been previously described to provide high levels of transgene expression in hepatocytes (see Nathwani AC, Gray JT, Ng CYC, Zhou J, Spence Y, Waddington SN, et al., Blood. 2006;107(7):2653-61.).

[0179] To generate this plasmid the human ABCB4 gene was codon optimised (SEQ ID NO: 44) and ordered from GeneArt with a 5’ Kozak sequence along 5’ Agel and 3’ Sall cutting sites. The DNA sequence was then cloned in a CCL. LP1. GFP backbone in place of the GFP gene. Upon confirmation of the cloning success by enzymatic digestion (Figure 2A) and sequencing (Figure 2B) the generated plasmid was used for producing the comparative CCL. LP1.coABCB4_v1 (SEQ ID NO: 28) by transient transfection of HEK293T cells with or without Verapamil (inhibitor of MDR3 function)(3). Titration of these vectors highlighted an inability of the CCL. LP1.coABCB4 (SEQ ID NO: 28) vector production in the absence of Verapamil, which for all concentrations increased the vector yield to above 1x104TU / ml (Figure 2. C). Despite this increase, the vector yield remained low requiring further optimisation.

[0180] CCL. LP1.coABCB4 treatment depolarises HepG2 cells in vitro

[0181] The vector CCL. LP1,coABCB4 comparative vector’s efficacy was also tested in vitro in ABCB4' HepG2 knock-out cells which could generate bile canaliculi-like structures (see Van derWouden JM, Van Uzendoorn SCD, Hoekstra D. Oncostatin M regulates membrane traffic and stimulates bile canalicular membrane biogenesis in HepG2 cells. EMBO Journal.

[0182] 2002;21(23):6409-18). The results showed that overexpression of the ABCB4 gene driven by the LP1 promoter leads to depolarisation of the treated cells and mislocalization of the MDR3 protein outside the bile canaliculi structures.

[0183] Example 2: Novel promoters for use in a lentiviral backbone

[0184] Novel promoters LPa and LPE improve titre and polarisation and are liver-specific

[0185] As demonstrated above, the comparative lentiviral vector CCL. LP1.coABCB4, comprising the known Liver Promoter 1 (LP1), had shown issues with poor titre and depolarisation with treated cells.

[0186] In order to resolve these issues, we generated new lentiviral vectors for PFIC3 carrying coABCB4 (SEQ ID NO: 9) under the control of different novel promoters. This included the hAAT promoter (SEQ ID NO: 40), the LPa promoter (SEQ ID NO: 3) or LPE promoter (SEQ ID NO: 45) (Figure 1 B). The hAAT promoter (SEQ ID NO: 40) was obtained by removing the HCR region (SEQ ID NO: 5) of the LP1 promoter in the comparative construct.

[0187] The novel LPa promoter (SEQ ID NO: 3) was engineered in silico. This was based on the natural ABCB4 promoter, however, while the predicted human ABCB4 promoter has a size of approximatively 1000bp, the size of LPa promoter (SEQ ID NO: 3) was reduced to 539bp to preserve only highly conserved regions and markers of promoters such as CpG islands from the natural sequence. Given this area overlapped with the first intron of the ABCB4 gene the splicing donor and splicing acceptor regions were mutated to 5’-CC-3’. This was paramount for ensuring safety of the vector. The novel LPE promoter (SEQ ID NO: 45) contains the LPa sequence (SEQ ID NO: 3) but also contains an enhancer region found upstream the natural ABCB4 promoter (SEQ ID NO: 66).

[0188] The novel LPa and LPE promoter sequences (SEQ ID NO: 3 and NO 45) were ordered from Geneart with flanking 5’ PspXI and 3’ Agel cutting sites and cloned instead of the known LP1 promoter as was used in comparative example CCL. LP1. coABCB4. This generated the example constructs: CCL. LPa. coABCB4 (SEQ ID NO: 29 - Version 2 coABCB4) and CCL. LPE. coABCB4 (SEQ ID NO: 30). These had a backbone corresponding to Figure 1B and may be referred to as a lentiviral vector with a Version 2 backbone herein.The sequence of the new transgene plasmids has been confirmed by sequencing (Figure 3 -A and B) and the constructs have been used to generate lentiviral vectors by transient transfection of HEK293T cells with 70mM Verapamil inhibition. Unexpectedly, CCL. LPa. coABCB4 (SEQ ID NO: 29 - Version 2 coABCB4) and CCL. LPE. COABCB4 (SEQ ID NO: 30 - Version 2 coABCB4) plasmids, both comprising the novel promoter, both produced increased vector titres compared to the comparative vector comprising the known promoter LP1 (Figure 3C).

[0189] Next, given that Example lentiviral vectors CCL. LPa. coABCB4 and CCL. LPE. COABCB4 had higher titres, we have tested in ABCB4~ / ~ HepG2 cells their efficacy in restoring MDR3 expression at bile canaliculi compared to the comparative lentiviral 1 vectors. As seen in Figure 3, D-F, both example vector constructs expressed ABCB4 at lower levels, which were closer to the physiological levels (Figure 3E). Whilst investigating the ability of HepG2 ABCB4'1' cells to polarise MDR3 at the bile canaliculi (Figure 3. G), we discovered that the CCL. LPa. coABCB4 (SEQ ID NO: 29) shows a similar polarisation pattern as wild type cells. Thus, the new construct not only increased the vector titers but allowed the MDR3 polarisation at the bile canaliculi compared to the comparative construct comprising the known promoter LP1.

[0190] To assess the in vivo biodistribution of the expressed example lentiviral vectors containing the novel LPa promoter we generated CCL. LPa. Luc. GFP (LPa, SEQ ID NO: 29 Version 2 GFP) reporter vector by replacing the coABCB4 gene (SEQ ID NO: 9) in the example construct with the Luciferase. GFP reporter genes (SEQ ID NO: 48). Three weeks old mice were then injected with 1x1010TU / kg LPa reporter vector. Two days post injections the mice were administered Luciferin intraperitoneally and imaged with an I VIS machine. As seen in Figure 4, A-B, the expression driven by the novel LPa promoter is localised to the liver and spleen. Encouraged by these results, the in vivo efficacy of the example vectors in rescuing the PFIC3 phenotype was assessed in neonate FVB.129P2-Abcb4fm76o7J mice injected with the example vectors CCL. LPa. coABCB4 (LV, SEQ ID NO: 29) or CCL. LPa. Luc. GFP (LG, SEQ ID NO: 31) and monitored for 18 weeks. The treatment showed an initial improvement in alanine aminotransferase levels at 6 weeks post treatment (Figure 3C). The data also confirmed the ability of the LPa promoter (SEQ ID NO: 3) to drive coABCB4 expression in the liver of treated animals.

[0191] Version 3 vectors produce sufficient titre and rescue the PFIC3 phenotype in vivo

[0192] The inventors sought to even further improve the vector such that there was increased likelihood of sustained efficacy of treatment. This is because a higher titre of therapeuticlentiviral vectors carrying the ABCB4 transgene may be needed if a low dose of vector is administered.

[0193] It was hypothesised that not only the function of ABCB4 but also its expression driven by the CMV promoter (Figure 1. A-B) in HEK293T cells may impact the vector titres. To prevent ABCB4 expression driven by the CMV promoter, we generated further new example lentiviral constructs with flipped transgene cassettes. These had a backbone corresponding to Figure 1C and D, and may otherwise be referred to as lentiviral vectors with a Version 3 backbone herein.

[0194] Example vectors include miR-coA4-LPaH (SEQ ID NO: 33 - coABCB4 Version 3, Figure 1D) and miR-wtA4-LPaH (SEQ ID NO: 34 - wtABCB4 Version 3, Figure 1D) and miR-GFP. Luc-LPaH (SEQ ID NO: 39). The new lentiviral constructs included 1) the reverse complement sequence of the LPa promoter (SEQ ID NO: 4) combined with the reverse complement of the HCR region (SEQ ID NO: 5) 3’ to the transgene to drive stable transgene hepatocyte expression, and 2) 4 x miR-126-3pT binding sites (SEQ ID NO: 11) and 4 x miR142 binding sites (SEQ ID NO: 13) to inhibit the post-transduction transgene expression in macrophages and dendritic cells.

[0195] To generate the new lentiviral vector backbone a fragment containing the reverse complement of bGH poly(A) signal (SEQ ID NO: 21) + 4x miR-126-3pT binding site and 4x miR142 binding site region (SEQ ID NO: 14) + reverse complement of WPRE (SEQ ID NO: 17) + Sall cutting site + reverse complement of GFP + Agel cutting site + LPaH promoter (SEQ ID NO: 6) was ordered from Geneart and cloned in CCL. LP1.coABCB4_v1 (SEQ ID NO: 28) between Xhol and Kpnl. The cloning success was confirmed by sanger sequencing (Figure 5A). Once confirmed, the miR-G-LPaH (SEQ ID NO: 32) was used for cloning the reverse complements of wild type ABCB4 (SEQ ID NO: 8), codon optimised ABCB4 gene (SEQ ID NO: 9) and Luciferase. GFP (SEQ ID NO: 49) to generate the miR-coA4-LPaH (SEQ ID NO: 33 - coABCB4 Version 3, Figure 1. D) and miR-wtA4-LPaH (SEQ ID NO: 34 - wtABCB4 Version 3, Figure 1D) and miR-GFP. Luc-LPaH (SEQ ID NO: 39). The success of the cloning was confirmed by Sanger sequencing (Figure 5 - B-D). The transgene plasmids were then used to produce lentiviral vectors through transient transfection of Hek293T cells without verapamil inhibition. Using the new lentiviral backbones above the titres of 1 E+8TU / ml were achieved for all the vectors with the highest titres obtained for miR-wtA4-LPaH (Figure 5F). T ransduction of HepG2 cells with miR-G-LPaH (SEQ ID NO: 32) vector confirmed the new construct could express the transgenes in target cells (Figure 5G).Given these findings, the efficacy of the example vector miR-wtA4-LPaH in rescuing the PFIC3 phenotype was tested in vivo in the F\ B. 29P2-Abcb4tm1Bor / J model (Figure 6). For this, neonate F\ / B. 29P2-Abcb4tm1Bor / J mice were pre-treated with 0.12g / kg intraperitoneal injections of clodronate liposomes at 24 and 6 hours before lentiviral vector administration to improve the lentiviral transduction efficacy (Touramanidou L et al, Cells [Internet]. 2024 Nov 29; 13(23): 1979. Available from: https: / / www.mdpi.com / 2073-4409 / 13 / 23 / 1979).

[0196] P3-4 mice were then injected with 3x1010TU / kg of miR-wtA4-LPaH (LV, n=6) and placed at 4 weeks for 9 days on a 0.10% CA diet to exacerbate the cholestatic phenotype along with WT (n=5) and untreated KO (n=6) animals (Figure 6A). The treated animals have shown improved survival post CA administration (Figure 6B), reduced weight loss on CA (Figure 6C) and highly reduced alanine transaminase (Figure 6D) and alkaline phosphatase levels at 5-6 weeks compared to untreated littermates (Figure 6E). At harvest, the treated animals also showed normalised bilirubin levels (Figure 6F and Figure 6G). Analysis of the liver vector copy number at harvest has shown an average of 5 vector copies in the liver of treated animals (Figure 6H). Importantly, picrosirius red staining for fibrotic tissue revealed that the liver of most treated mice has minimal fibrosis, similar to the one found in wild type animals (Figure 6I and 6J). As measured by mass spectrometry the miR-wtA4-LPaH treatment also restored the excretion of the phosphatidyl choline into the bile (Figure 7A) and the expression of ABCB4 protein in the liver tissue (Figure 7B) These, support the efficacy of the example vector miR-wtA4-LPaH in rescuing the PFIC3 phenotype in the FVB.129P2-Abcb4fm76o7J PFIC3 mouse model.

[0197] Version 3 vectors have shown safe integration patterns in primary human hepatocytes

[0198] To assess the safety of the vector in a cellular model relevant for humans, primary human hepatocytes sourced from BIOIVT were plated per the manufacturer’s instructions into 12-well plates at a density of 700000 cells / well. After 24h in culture the cells were transduced with miR-wtA4-LPaH (SEQ ID NO: 34) lentiviral vector at a multiplicity of infection of 2 and 20. The cells were harvested 72h post transduction and their DNA was sent for integration site analysis to assess the integration pattern in these cells. Figure 8 shows results of integration site analysis. Cells treated with the miR-wtA4-LPaH (SEQ ID NO: 34) at both low and high MOI show signs of polyclonal and oligoclonal integration with Shannon diversity scores generally above 0.8 (Figure 8A) and diverse integration sites (Figure 8B). Cells showed similar total integration numbers (Figure 8C). Circle packing plots show the genes with Gene Relative Abundance (GRA) greater than 0.0001, with larger circles showing higher frequency of integration (Figure 8D-E). Representative plot of one replicate treated with MOI 2 (Figure 8D) and one replicated treated with MOI 20 (Figure 8E). The genes with top 5 integration sites ineach of the replicates are not known oncogenes (Figure 8F). This indicated that the vector is likely to integrate in safe genetic locations in primary human hepatocytes.

[0199] Comparative Example 3 - Lentiviral Vector - CCL. LP1.coABCB11

[0200] A CCL. LP1.coABCB11 lentiviral vector backbone comprising the known promoter LP1 impedes vector production

[0201] We aimed to develop and test a liver directed lentiviral gene therapy for treating PFIC2. For this, a self-inactivating lentiviral vector, named CCL. LP1. coABCBI 1 (SEQ ID NO: 69 -CCL. LP1.coABCB11_v1, Figure 1), carrying the codon optimised ABCB11 (SEQ ID NO: 71) under the control of a known liver-specific Liver Promoter 1 (LP1, SEQ ID NO: 42) was first generated. The LP1 promoter (SEQ ID NO: 42) composed of the human apolipoprotein hepatic control region (HCR, SEQ ID NO: 41) and the human alpha-1-antitrypsin (hAAT) promoter (SEQ ID NO: 40) (20) was chosen for initial experiments as it has been previously described to provide high levels of transgene expression in hepatocytes (see Nathwani AC, Gray JT, Ng CYC, Zhou J, Spence Y, Waddington SN, et al., Blood. 2006;107(7):2653-61.).

[0202] To generate this plasmid the human ABCB11 gene was codon optimised (SEQ ID NO: 71) and ordered from GeneArt with a 5’ Kozak sequence along 5’ Agel and 3’ Sall cutting sites. The DNA sequence was then cloned in a CCL. LP1. GFP (SEQ ID NO: 74) backbone in place of the GFP gene. The generated plasmid was used for producing the comparative CCL. LP1.coABCB11_v1 (SEQ ID NO: 69) by transient transfection of HEK293T cells. Titration of these vectors highlighted a reduced ability of the CCL. LP1.coABCB11 (SEQ ID NO: 69) to generate lentiviral particles compared to the CCL. LP1. GFP (GEW, SEQ ID NO: 74) (Figure 9A). The CCL. LP1.coABCB11_v1 (SEQ ID NO: 69) expresses the ABCB11 protein in Hek293T cells during viral production which likely interfere with the vector packaging (Figure 9B).

[0203] Example 4 - Version 3 vectors containing the coABCBH gene do not express the ABCB11 protein in producer Hek293T cells

[0204] To generate the miR-coABCB11-LPaH vector (SEQ ID NO: 70 - miR-coABCB77-LPaH, Version 3 backbone - coABCBH) the reverse complement of the coABCBH gene (SEQ ID NO: 73 - Reverse complement of codon-optimised ABCB11) was cloned in place of the GFP in the miR-G-LPaH (SEQ ID NO: 32) construct. The new plasmid was the used to transfect Hek293T cells. Western blotting has shown this construct does not allow expression of the ABCB11 protein in the producer cells (Figure 9B).Discussion

[0205] PFIC3 is a severe autosomal recessive disorder which leads to progressive liver cholestasis, fibrosis and end stage liver disease. With no curative treatments, most patients require liver transplantation which is often challenging. Several gene therapy approaches such as mRNA tor AAV gene therapy have been investigated for this disease however they face several challenges such as limited efficacy over time. Lentiviral gene therapy would be an ideal treatment for this disorder. However, the development of lentiviral vectors for PFIC3 has previously been hindered by the expression of the ABCB4 in the HEK293T producer cells. These findings were in alignment with previous observations which stated that the development of lentiviral gene therapy for PFIC would be challenging (van Til NP et al, Retrovirology. 2008;5:1-9.). Additionally, overexpression of ABCB4 in target cells was also observed to be problematic.

[0206] Here, we have detailed a stepwise process through which we developed novel lentiviral promoters and novel lentiviral vector constructs for PFIC3 capable of producing high level titres, and which showed improved results compared to known liver promoters LP1.

[0207] The preferred new backbone conformation (Figure 1 - C-D) importantly supresses the expression of the ABCB4 transgene in producer cells by reverting the expression cassette in the lentiviral vector genome, removing the translation starting site from the positive sense RNA. During the reverse transcription process of the lentiviral vector a double stranded DNA is generated and integrated in the target genome. This DNA will allow the expression of the transgene of interest (i.e. ABCB4) from the internal promoter (i.e. LPa) included in the construct. Using this technology the therapeutic gene is expressed only in target cells but not in the producer cells. It is understood that such constructs can be used to block the expression of transgenes of interest in producer cells during lentiviral vector production.

[0208] We have also shown that PFIC3 phenotypic correction could be achieved with the novel lentiviral vectors in FVB.129P2-Abcb4fm76o7J PFIC3 murine disease model. In our preliminary studies, treating FVB.129P2-Abcb4“7J mice with the miR-wtA4-LPaH (SEQ ID NO: 33) carrying the reverse complement of wild type ABCB4 (SEQ ID NO: 8) and a chimeric promoter containing our novel LPa promoter (SEQ ID NO: 4) has shown improvement of several PFIC3 specific parameters.Overall, these results not only are relevant for therapies for PFIC3 patients but also for other patients suffering from genetic disorders amenable to lentiviral gene therapy, but for whom the production of lentiviral vectors is hindered by the therapeutic gene expression in producer cells such as for PFIC2.

[0209] Sequences

[0210] Sequence Elements

[0211] SEQ ID NO: 1 LPa promoter gggtggcccatcaggcgaggctttctcggcacccgaggctccagcctgatctcggtcttatcctgcgaccgggctggttctggcgg gtcgccagggtgggcggcggccccagccgggcgccccggcggcaagagcggcaggctgcgcccctggcccgcgcctagcc tggggagagagctgggcgggcggcgggagctgctctcgcgggccgcggccctcgccctggctgcaacggtaggcgtttcccg ggccggacgcgcgtggggggcgggggcgggggcgggggcgaggccgcggcgagcaaagtccaggcccctctgctgcagc gcccgcgcgtccagaggccctgccagacacgcgcgaggttcgagnngagagaggtccgggcgcgtctggcctcgaaggga gacccgggacgtggggcgcggggcgggagtggccggacctccacccagtgcccccgggccccgcgactcgtgcgccgggc cgccggagagggtgtacttggttctgaggctgtggtttctcctcnngctgag

[0212] Mutated splice sites are shown in bold.

[0213] SEQ ID NO: 2 LPa promoter reverse complement ctcagcnngaggagaaaccacagcctcagaaccaagtacaccctctccggcggcccggcgcacgagtcgcggggcccgg gggcactgggtggaggtccggccactcccgccccgcgccccacgtcccgggtctcccttcgaggccagacgcgcccggacct ctctcnnctcgaacctcgcgcgtgtctggcagggcctctggacgcgcgggcgctgcagcagaggggcctggactttgctcgccg cggcctcgcccccgcccccgcccccgccccccacgcgcgtccggcccgggaaacgcctaccgttgcagccagggcgaggg ccgcggcccgcgagagcagctcccgccgcccgcccagctctctccccaggctaggcgcgggccaggggcgcagcctgccgc tcttgccgccggggcgcccggctggggccgccgcccaccctggcgacccgccagaaccagcccggtcgcaggataagaccg agatcaggctggagcctcgggtgccgagaaagcctcgcctgatgggccaccc

[0214] Mutated splice sites are shown in bold.

[0215] SEQ ID NO: 3 LPa promoter as used in Examples gggtggcccatcaggcgaggctttctcggcacccgaggctccagcctgatctcggtcttatcctgcgaccgggctggttctggcgg gtcgccagggtgggcggcggccccagccgggcgccccggcggcaagagcggcaggctgcgcccctggcccgcgcctagcc tggggagagagctgggcgggcggcgggagctgctctcgcgggccgcggccctcgccctggctgcaacggtaggcgtttcccg ggccggacgcgcgtggggggcgggggcgggggcgggggcgaggccgcggcgagcaaagtccaggcccctctgctgcagc gcccgcgcgtccagaggccctgccagacacgcgcgaggttcgagccgagagaggtccgggcgcgtctggcctcgaaggga gacccgggacgtggggcgcggggcgggagtggccggacctccacccagtgcccccgggccccgcgactcgtgcgccgggc cgccggagagggtgtacttggttctgaggctgtggtttctcctcccgctgag

[0216] Mutated splice sites are shown in bold

[0217] SEQ ID NO: 4 Lpa promoter reverse complement as used in Examples ctcagcgggaggagaaaccacagcctcagaaccaagtacaccctctccggcggcccggcgcacgagtcgcggggcccgg gggcactgggtggaggtccggccactcccgccccgcgccccacgtcccgggtctcccttcgaggccagacgcgcccggacct ctctcggctcgaacctcgcgcgtgtctggcagggcctctggacgcgcgggcgctgcagcagaggggcctggactttgctcgccg cggcctcgcccccgcccccgcccccgccccccacgcgcgtccggcccgggaaacgcctaccgttgcagccagggcgaggg ccgcggcccgcgagagcagctcccgccgcccgcccagctctctccccaggctaggcgcgggccaggggcgcagcctgccgc tcttgccgccggggcgcccggctggggccgccgcccaccctggcgacccgccagaaccagcccggtcgcaggataagaccg agatcaggctggagcctcgggtgccgagaaagcctcgcctgatgggccaccc

[0218] Mutated splice sites are shown in bold

[0219] SEQ ID NO: 5 - HCR enhancer cctctcacactacctaaaccacgccaggacaacctctgctcctctccaccgaaattccaaggggtcgagtggatgttggaggtgg catgggcccagagaggtctctgacctctgccccagctccaaggtcagcaggcagggagggctgtgtgtttgctgtttgctgcttgca atgtttgcccattttaggg

[0220] SEQ ID NO: 6 - Lpa promoter + HCR enhancerctcagcnngaggagaaaccacagcctcagaaccaagtacaccctctccggcggcccggcgcacgagtcgcggggcccgg gggcactgggtggaggtccggccactcccgccccgcgccccacgtcccgggtctcccttcgaggccagacgcgcccggacct ctctcnnctcgaacctcgcgcgtgtctggcagggcctctggacgcgcgggcgctgcagcagaggggcctggactttgctcgccg cggcctcgcccccgcccccgcccccgccccccacgcgcgtccggcccgggaaacgcctaccgttgcagccagggcgaggg ccgcggcccgcgagagcagctcccgccgcccgcccagctctctccccaggctaggcgcgggccaggggcgcagcctgccgc tcttgccgccggggcgcccggctggggccgccgcccaccctggcgacccgccagaaccagcccggtcgcaggataagaccg agatcaggctggagcctcgggtgccgagaaagcctcgcctgatgggccaccccctcgaggcctctcacactacctaaaccacg ccaggacaacctctgctcctctccaccgaaattccaaggggtcgagtggatgttggaggtggcatgggcccagagaggtctctg acctctgccccagctccaaggtcagcaggcagggagggctgtgtgtttgctgtttgctgcttgcaatgtttgcccattttaggg Mutated splice sites are shown in bold.

[0221] SEQ ID NO: 7- Reverse complement of LPa promoter + HCR enhancer ctcagcgggaggagaaaccacagcctcagaaccaagtacaccctctccggcggcccggcgcacgagtcgcggggcccgg gggcactgggtggaggtccggccactcccgccccgcgccccacgtcccgggtctcccttcgaggccagacgcgcccggacct ctctcggctcgaacctcgcgcgtgtctggcagggcctctggacgcgcgggcgctgcagcagaggggcctggactttgctcgccg cggcctcgcccccgcccccgcccccgccccccacgcgcgtccggcccgggaaacgcctaccgttgcagccagggcgaggg ccgcggcccgcgagagcagctcccgccgcccgcccagctctctccccaggctaggcgcgggccaggggcgcagcctgccgc tcttgccgccggggcgcccggctggggccgccgcccaccctggcgacccgccagaaccagcccggtcgcaggataagaccg agatcaggctggagcctcgggtgccgagaaagcctcgcctgatgggccaccccctcgaggcctctcacactacctaaaccacg ccaggacaacctctgctcctctccaccgaaattccaaggggtcgagtggatgttggaggtggcatgggcccagagaggtctctg acctctgccccagctccaaggtcagcaggcagggagggctgtgtgtttgctgtttgctgcttgcaatgtttgcccattttaggg SEQ ID NO: 8 - Reverse complement of wild-type ABCB4 TCATAAGTTCTGTGTCCCAGCCTGGACACTGACCATTGAAAAATAGATGCCTTTCTGTGC CAGCAGCTGCTGATGCGTGCCATGCTCCTTGACTCTCCCATTCTGAAACACCACTATTA AGTCTGCATTCTGGATGGTGGACAGGCGGTGAGCAATCACAATGCAGGTGCGGCCTTC TCTGGCTTTGTCCAGGGCTTCTTGGACAACCTTTTCACTTTCAGTATCCAGAGCTGATGT AGCTTCATCCAACAGGAGGATTTGAGGTTGTCTGATGAGGGCTCGGGCAATAGCAATCC TCTGTTTTTGACCTCCTGAGAGCTGAGTCCCCTTATCTCCCACTCTTGTTTCATATTTGT GGGGTAACGTCTCGATGAAAGGATGTATGTTGGCAGCTTTGGCTGCACTCACAATTTCA TCCTGTGATACAACCCGGCTGTTGTCTCCATAGGCAATATTCTCGGCAATGCTGCAGTC AAATAGGATAGGCTCCTGAGACACGATTCCGAGTTGAGCTCTGAGCCACTGGACATTGA GTTTCTTTGCTTCTTGACCATCGAGAAGCACTGTCCCCGCCAAGGGGTCGTAGAACCGC TCCAGGAGCTGGACCACCGTGCTCTTCCCACAGCCACTGCTGCCCACCAGGGCTAGTG TCTGGCCTTTCTTCACCTCCAGGCTCAGCCCCTGAAGCACTGGCACGTTTGCTCGGGT GGGATAGTTGAACACGACTTCATTAAATGTTATATTTCCTTCAAATTTATCAGGCTTCAGC CCCTCTTCACTGTAGCTGTCAATCAGAGGTTGTCTTTCAAACAGCATGAATAAGTGGGCT GCAGACAGCTTAGCTTTAGCATAGTCTGGAGCAAATGAACTGGCATGTCCTAGAGCCAC TGCACCAAATACAATTGCAGAAAACACCAGAATAACATCTCTGAAGCGCATATGTCCATT CACAATGAGATATGCACCAAATCGAAAACAACCGGCATAGGAAAAATACATAAATGCTT GTGAGATACTAAAAGTAATTCCATAGATGTGTGCCTTCTGCACAGAATTCCTGTAAGGTC CATACAATTTTTCAACATACATTGATTCAAATTTTCTTTCCTGGGTCAAAGACACAACTGT CCTAATATTTTCTATTGCCTCTGTTGCAATCTTTCCAGCAGCTTCCAGTTCTTTTTTATCT CTTTTGGCATTTCCAGCCAACAATTTCATTTCAACAATTCCTGACACAGCAATAATTGGAA CAACTGCTAATAGCAATAGGGTTAACTGCCAACCGTAGATAAATGATATGATAATACCAG TTCCAAGGTTAGCTATATTCTGTGCAATTAAAGCCAACCTGGTTCCTGTGGCTCCTTGGA CTTGGGCAGCATCTGTGGCAAGTCTTGTAGAAAGTGCACCAGTACTGTTTTTATGGTCA TCAAACCAGCTCATGTCCTGTCTTAGCATTGCTTTAAAAGCCATTGACCGCAGTCTTCTG GTGAGGATCTCGCCAGCTTTCCCAAACGTGAAACCCTGAAGGAAGAAAGTAAAAAAAGA AATAATTCCCAGAAATAAGAAAATCAAAGAGAATATGTTGCACTTCTGCTGCTTCACTGC ATCATCGCCTGGTCCAAAAATCGCTATGATCTCTGAGAATATGACTGAAAATGCCGGCT GAAGCCCCCCATTGGCAATGGCACATACTGTTCCCACGACAAAGTAGGGCCATTCTGTT TTATTCAGTTTCAGGACCTTCAGAAAGGACACTGGTGGCACATTTGCTTCAAGTCCATCG GTTTCCACATCAAGGCTCTTCTGACACATTTGTGAATTTTTAAGGTTTTTCTGAGTAGAAT GCCTAAATAGGCGAGATTTCCAGCCATTTGGGGCCATTCTAGTGGCAGCCTTTTCATCATTTAGTTCAAATTCTTCTGACTGGATCTGGCTTCCTGATGTCTGCATGTTGACAAGTTTG AAGTACACCCCTTCCTTCTTCATCAGTTCGCTGTGGCTTCCTTGCTCCACAATTACTCCA TCCTCAAACCCAGCGATGACATCTGCATTTCGGACCGTAGACAGTCGGTGTGCTATCAC AATGGTGGTCCGGCCTTCTCTGGCCTTATCCAGAGCTGCCTGTACCTCAGCTTCACTTT CTGTGTCCAATGCTGACGTGGCCTCATCCAGCAGAAGGATCTTGGGGTTGCGAACCAG GGCACGTGCAATGGCGATCCTCTGCTTCTGCCCACCACTCAGCTGGGCCCCTCTCTCT CCAACCAGGGTGTCAAATTTCTGTGGTAATTTCATGATAAACTCATAGGCGTTGGCCTCT TTGACAGCTTTCTTTATCTCATCCATGGTTACATTTCCACGGCCATAACAAATATTTTCAG CAATTGTGGTGGAAAACAGCACCGGCTCCTGACTCACCACACCAATGATTTCCCTCAGA TAGTTTACATTAAAGTTCCTAATATCCTGCCCATCAATGTTAATTGTGCCCTCATCAGGGT CATAGAGCCTCTGTATCAGCTGGACCGTTGTGCTCTTCCCACAGCCACTACTTCCAACC AGGGCCACCGTCTGCCCACTCTGCACCTTCAGGTTGAGGCCCTTCAAGATCTTGACGTT AGCTCGAGAAGGGTAAGAAAAGTGAACATCATTGAACTCCAAATTCCCTTTGATGCTGT CTGGTTTGTGTCCTCTCTCTGAAAAACTGTCAATTTTAGGATTATTATCAATAATATCAAA GATCACATATGCTGCTCCTCTTGCATTGGCAAAAGCATCAATACATGGGGCAGCCTGGC CAACACTGAAAGCTCCAATTAGGATTGAAAAAAAAACTGTCATTGCATTTCCAATAGTAT ATTCTTTTGATATGACTAGAGTGGATCCATACCAGAAGGCCAGTGCATATGATGCATATA TTAACAGGAAGGCAATACCCATGGAAATGTTTGCTGAAATAGCTTTTTTAATTCCAATCT CTTTGGCATTTTCTAAATGTTTCTGATACCTTTCCAGCTCTTTGTTCTGGCCCCCGAAAG CTATCACAGTCCTGATGGCCCCCAGAGCCTCTTCTGCCACGGCGCCTGCTTTTGCATAA GCAGCTAGTTCTTTGTCACTAAATGCCGAGAGTATCTTTGCCCAAACGGCTGCAGAGAG TCCTAGAATAGGGCTGATGGCCATTATCACAAGGGTGAGCTTCCATCCTCTGATGAATC CCACTATGAATCCTGCAAAAAACGTGGCTACTGCTTGAAAGAACATTCCAACCTTGTCAC CAATTCCTTCACTGATTTTGGAGATGTCATCTGTTAGCCGCGTATTGAGTTCAGTGGTGT CGTTGATGTCAAACCATCCTATTTCCTGTCGTAGAATAGCATGAAAAAACTTCTGCCTAA TTTTCCTGATCTGCCGACCAGCTGCCAAAGTCCAAAATGAAACTTGTATATAGGCAGCAA CAAGAACTCCAGCACCCAATCCTGAGTAGTAATATGCATATCTAGTCATTTCTTCTTCCA GAATTTTGCCTGGATTTAGCAGCGACAAGGAAAAGTTCACTGGAAAGGAGAAGTTTCCT GCAGTATCAACAAATTTGTCAGTCATCTCTCCAAATACTATCATCATGAGGGGGAGACCT GATCCGTGAGCTATGGCCATGATGGTACCCAGCGACATAAACAATTTATCCTGCCAATC GGAGTATCGAAACAATGTTAATACTCCAATCATTTTCACTGTCTTCGTTTTTTTCCTTTTTT GTTTGCTGCTGATGCCCAGTTCAAAGTCGCCCTCCGCGCTCGTGGGGCGCCAGGCTGT TCCGTTCTTTGCCGCCTCAAGATCCAT SEQ ID NO: 9 - Reverse complement of codon-optimised ABCB4 TCACAGGTTCTGTGTGCCGGCCTGCACAGACACCATGGAGAAGTAGATGCCCTTCTGG GCCAGCAGTTGCTGGTGGGTGCCGTGCTCTTTCACTCTGCCGTTCTGGAACACCACGA TGAGGTCGGCATTCTGGATGGTGCTCAGCCGGTGAGCGATCACGATACAGGTTCTTCC CTCGCGGGCCTTATCCAGAGCCTCTTGCACCACCTTCTCGCTCTCGGTGTCCAGAGCA GAGGTGGCCTCATCCAGGAGCAGAATCTGAGGCTGTCTGATCAGGGCGCGAGCGATA GCAATCCTCTGTTTCTGGCCTCCGCTCAGCTGTGTGCCCTTGTCGCCCACTCTTGTCTC GTATTTGTGGGGCAGTGTCTCGATGAAGGGGTGAATGTTGGCAGCCTTGGCAGCGGAC ACAATCTCGTCCTGGGACACGACTCTGCTGTTGTCGCCATAGGCAATATTCTCGGCAAT GCTGCAGTCGAACAGGATGGGTTCTTGAGACACGATTCCCAGCTGAGCCCTCAGCCAC TGCACGTTCAGCTTTTTGGCTTCTTGGCCATCCAGCAGGACGGTTCCAGCCAGAGGATC GTAGAACCGTTCCAGCAGCTGCACCACTGTAGACTTGCCACATCCGCTAGAGCCCACG AGAGCCAGAGTCTGGCCCTTCTTCACTTCCAGGCTCAGTCCCTGCAGAACTGGCACGTT GGCTCTGGTGGGGTAGTTGAACACCACCTCGTTAAAGGTGATGTTGCCCTCGAACTTAT CTGGCTTCAGGCCTTCCTCGCTGTAGGAGTCGATCAGGGGCTGTCTCTCGAACAGCAT GAACAGGTGTGCGGCAGACAGCTTGGCCTTGGCGTAGTCAGGGGCAAAGCTAGAGGC ATGTCCCAGAGCCACGGCTCCAAACACGATGGCGGAGAACACCAGGATCACGTCCCGG AATCTCATGTGGCCGTTCACGATCAGGTAGGCGCCGAATCTAAAGCAGCCGGCGTAGC TAAAGTACATGAAGGCCTGGCTGATGCTGAAGGTGATGCCGTAGATGTGGGCCTTCTG CACGCTGTTTCTGTAGGGGCCGTACAGCTTTTCCACGTACATAGACTCGAACTTCCGCT CTTGGGTCAGAGACACCACGGTCCGGATATTCTCGATGGCCTCGGTGGCGATCTTGCCTGCGGCCTCAAGTTCTTTCTTGTCCCGCTTGGCGTTTCCGGCCAGCAACTTCATTTCCA CGATGCCGGACACGGCAATGATAGGCACCACGGCCAGCAGGAGCAGTGTCAGCTGCC ATCCGTAGATGAAGCTAATGATGATGCCGGTGCCGAGATTGGCGATATTCTGGGCGATC AGGGCCAGTCTTGTGCCTGTAGCGCCTTGCACCTGAGCAGCATCTGTGGCCAGTCTGG TAGACAGGGCGCCAGTGCTGTTCTTGTGGTCGTCGAACCAACTCATGTCCTGTCTCAGC ATGGCTTTGAAGGCCATGGATCTCAGTCTCCGAGTCAGGATCTCCCCGGCCTTGCCGA AGGTGAAGCCTTGCAGGAAGAATGTAAAGAAGCTGATAATCCCCAGGAACAGGAAGAT CAGGCTAAAGATATTACACTTCTGCTGCTTCACGGCGTCGTCGCCAGGGCCAAAGATG GCGATAATCTCGCTAAAGATCACGGAGAAGGCGGGCTGCAGTCCACCATTAGCAATGG CACACACGGTGCCCACGACGAAGTAGGGCCACTCGGTCTTGTTCAGTTTCAGCACTTTC AGGAAGGACACAGGAGGCACATTGGCTTCCAGTCCGTCTGTCTCGACGTCCAGGGACT TCTGGCACATCTGGCTGTTCTTCAGATTCTTCTGGGTGCTGTGTCTGAACAGCCGGGAC TTCCAGCCATTAGGGGCCATCCTGGTGGCGGCCTTCTCGTCATTCAGCTCGAATTCCTC GCTCTGGATCTGAGAGCCGCTGGTCTGCATGTTGACCAGCTTGAAGTACACGCCTTCTT TCTTCATCAGCTCGCTGTGGGAGCCCTGTTCCACGATCACGCCATCTTCGAATCCGGCG ATCACATCGGCATTCCGCACGGTGGACAGTCTGTGGGCAATCACGATGGTGGTTCTGC CCTCTCTAGCCTTGTCCAGGGCAGCCTGCACTTCTGCCTCAGACTCTGTATCCAGGGC GCTTGTGGCTTCGTCCAGCAGCAGGATCTTAGGGTTCCGCACGAGTGCTCTGGCAATG GCGATTCTCTGCTTCTGTCCGCCAGACAGTTGGGCGCCTCTTTCTCCCACAAGGGTGTC GAATTTCTGGGGCAGCTTCATGATGAACTCGTAGGCGTTGGCCTCTTTCACGGCTTTCT TGATCTCGTCCATGGTCACGTTGCCTCTGCCGTAGCAGATGTTCTCGGCGATTGTGGTG CTGAACAGCACAGGCTCTTGGGACACCACTCCGATGATCTCGCGCAGGTAGTTCACGT TGAAGTTCCGGATGTCCTGGCCGTCGATGTTGATTGTGCCCTCGTCAGGGTCGTACAGT CTCTGGATCAGCTGCACGGTGGTGCTCTTGCCACAGCCAGAAGATCCCACGAGGGCCA CTGTCTGTCCGCTCTGCACCTTCAGGTTCAGGCCCTTCAGGATCTTCACGTTGGCCCGG CTAGGGTAGCTGAAGTGCACGTCGTTGAATTCCAGGTTGCCCTTGATGCTATCAGGCTT GTGGCCCCGCTCGCTGAAGCTGTCGATCTTGGGGTTGTTGTCGATGATATCGAAGATCA CGTATGCGGCGCCTCTGGCATTAGCAAAGGCGTCGATACAAGGAGCGGCCTGTCCGAC GCTAAAGGCGCCGATCAGGATGGAGAAGAACACGGTCATGGCGTTGCCGATGGTGTAC TCTTTGCTGATGACCAGGGTAGAGCCGTACCAAAAGGCCAGGGCGTAGCTGGCGTAGA TCAGCAGGAAGGCGATTCCCATAGAGATGTTGGCGCTGATGGCCTTCTTGATGCCAATC TCTTTGGCGTTCTCGAGGTGCTTCTGGTATCTTTCCAGTTCTTTGTTCTGGCCGCCAAAG GCGATCACGGTTCTGATGGCTCCCAGGGCTTCTTCAGCAACGGCGCCAGCTTTAGCGT AAGCGGCCAGCTCTTTGTCGCTGAAGGCGCTCAGAATCTTGGCCCACACAGCAGCAGA CAGTCCCAGGATGGGAGAGATGGCCATGATCACGAGGGTCAGCTTCCAGCCTCTGATG AAGCCCACGATAAAGCCGGCGAAGAAGGTGGCCACGGCCTGGAAGAACATTCCCACTT TGTCGCCGATGCCTTCGCTGATCTTGCTGATGTCGTCGGTCAGTCTGGTGTTCAGCTCG GTGGTGTCGTTGATGTCGAACCATCCGATCTCTTGCCGCAGGATGGCGTGGAAGAACT TCTGCCGGATCTTTCTGATCTGCCGGCCAGCGGCCAGTGTCCAAAAGGACACCTGGAT ATAGGCGGCCACCAGAACGCCAGCTCCAAGGCCGCTGTAGTAGTAGGCGTATCTGGTC ATTTCCTCTTCCAGGATCTTGCCGGGGTTCAGCAGGCTCAGGCTGAAATTCACGGGGAA GCTGAAGTTGCCGGCGGTGTCCACGAACTTGTCGGTCATCTCGCCGAACACGATCATC ATCAGGGGCAGTCCAGATCCGTGGGCGATAGCCATGATGGTGCCCAGGCTCATGAACA GCTTGTCCTGCCAGTCGCTGTACCGGAACAGGGTCAGCACGCCGATCATCTTCACGGT CTTGGTCTTTTTCCGCTTCTGCTTGCTGCTGATGCCCAGCTCAAAATCGCCCTCGGCAG ATGTAGGCCTCCAAGCGGTGCCATTCTTGGCGGCTTCCAGATCCAT SEQ ID NO: 10 - miR-126-3pT binding site

[0222] tcgtaccgtgagtaataatgcg

[0223] SEQ ID NO: 11 - 4 x miR-126-3pT binding site TCGTACCGTGAGTAATAATGCGATCGTCGTACCGTGAGTAATAATGCGCGGTTCGTACC GTGAGTAATAATGCGATCGTCGTACCGTGAGTAATAATGCG SEQ ID NO: 12 - 1 x miR-142 binding siteTGTAGTGTTTCCTACTTTATGGA

[0224] SEQ ID NO: 13 - 4 x miR142 binding site TGTAGTGTTTCCTACTTTATGGAGTGATGTAGTGTTTCCTACTTTATGGACGGTTGTAGT GTTTCCTACTTTATGGAATCGTGTAGTGTTTCCTACTTTATGGA SEQ ID NO: 14 - 4x miR-126-3pT binding site + 4x miR142 binding site region TCGTACCGTGAGTAATAATGCGATCGTCGTACCGTGAGTAATAATGCGCGGTTCGTACC GTGAGTAATAATGCGATCGTCGTACCGTGAGTAATAATGCGCTGCAGGTGTAGTGTTTC CTACTTTATGGAGTGATGTAGTGTTTCCTACTTTATGGACGGTTGTAGTGTTTCCTACTTT ATGGAATCGTGTAGTGTTTCCTACTTTATGGA SEQ ID NO: 15 (LTR-LTR sequence of example construct comprising wt ABCB4) GGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGGCTAACTAGGGAACC CACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCCCGTCTG TTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAAAATCTCT AGCAGTGGCGCCCGAACAGGGACTTGAAAGCGAAAGGGAAACCAGAGGAGCTCTCTC GACGCAGGACTCGGCTTGCTGAAGCGCGCACGGCAAGAGGCGAGGGGCGGCGACTG GTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGAAGGAGAGAGATGGGTGCGAGA GCGTCAGTATTAAGCGGGGGAGAATTAGATCGCGATGGGAAAAAATTCGGTTAAGGCC AGGGGGAAAGAAAAAATATAAATTAAAACATATAGTATGGGCAAGCAGGGAGCTAGAAC GATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGTAGACAAATACTGGGA CAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATCATTATATAATACAGTA GCAACCCTCTATTGTGTGCATCAAAGGATAGAGATAAAAGACACCAAGGAAGCTTTAGA CAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCACAGCAAGCGGCCGCTGAT CTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAGTGAATTATATAAATATA AAGTAGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGCAAAGAGAAGAGTGGTG CAGAGAGAAAAAAGAGCAGTGGGAATAGGAGCTTTGTTCCTTGGGTTCTTGGGAGCAG CAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACAGGCCAGACAATTATT GTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTGAGGCGCAACAGCAT CTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAGAATCCTGGCTGTGG AAAGATACCTAAAGGATCAACAGCTCCTGGGGATTTGGGGTTGCTCTGGAAAACTCATT TGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTCTGGAACAGATTTG GAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTACACAAGCTTAATAC ACTCCTTAATTGAAGAATCGCAAAACCAGCAAGAAAAGAATGAACAAGAATTATTGGAAT TAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACATAACAAATTGGCTGTGGTATATAA AATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTTTTGCTGTACTTTC TATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGACCCACCTCCCAAC CCCGAGGGGACCCGACAGGCCCGAAGGAATAGAAGAAGAAGGTGGAGAGAGAGACAG AGACAGATCCATTCGATTAGTGAACGGATCTCGACGGTATCGGTTAACTTTTAAAAGAAA AGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGACATAATAGCAACAGAC ATACAAACTAAAGAATTACAAAAACAAATTACAAAAATTCAAAATTTTATCGATCACGAGA CTAGCCTCGAGGATATCCCATAGAGCCCACCGCATCCCCAGCATGCCTGCTATTGTCTT CCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCAGAATAGAATGACACCTA CTCAGACAATGCGATGCAATTTCCTCATTTTATTAGGAAAGGACAGTGGGAGTGGCACC TTCCAGGGTCAAGGAAGGCACGGGGGAGGGGCAAACAACAGATGGCTGGCAACTAGA AGGCACAGACGCGTGGGCCCCTGCAGTCGTACCGTGAGTAATAATGCGATCGTCGTAC CGTGAGTAATAATGCGCGGTTCGTACCGTGAGTAATAATGCGATCGTCGTACCGTGAGT AATAATGCGCTGCAGGTGTAGTGTTTCCTACTTTATGGAGTGATGTAGTGTTTCCTACTT TATGGACGGTTGTAGTGTTTCCTACTTTATGGAATCGTGTAGTGTTTCCTACTTTATGGA GGCTAGCCTAGGCGGGGAGGCGGCCCAAAGGGAGATCCGACTCGTCTGAGGGCGAAG GCGAAGACGCGGAAGAGGCCGCAGAGCCGGCAGCAGGCCGCGGGAAGGAAGGTCCG CTGGATTGAGGGCCGAAGGGACGTAGCAGAAGGACGTCCCGCGCAGAATCCAGGTGG CAACACAGGCGAGCAGCCATGGAAAGGACGTCAGCTTCCCCGACAACACCACGGAATT GTCAGTGCCCAACAGCCGAGCCCCTGTCCAGCAGCGGGCAAGGCAGGCGGCGATGAGTTCCGCCGTGGCAATAGGGAGGGGGAAAGCGAAAGTCCCGGAAAGGAGCTGACAGGT GGTGGCAATGCCCCAACCAGTGGGGGTTGCGTCAGCAAACACAGTGCACACCACGCCA CGTTGCCTGACAACGGGCCACAACTCCTCATAAAGAGACAGCAACCAGGATTTATACAA GGAGGAGAAAATGAAAGCCATACGGGAAGCAATAGCATGATACAAAGGCATTAAAGCA GCGTATCCACATAGCGTAAAAGGAGCAACATAGTTAAGAATACCAGTCAATCTTTCACAA ATTTTGTAATCCAGAGGTTGATTGCTCATAAGTTCTGTGTCCCAGCCTGGACACTGACCA TTGAAAAATAGATGCCTTTCTGTGCCAGCAGCTGCTGATGCGTGCCATGCTCCTTGACT CTCCCATTCTGAAACACCACTATTAAGTCTGCATTCTGGATGGTGGACAGGCGGTGAGC AATCACAATGCAGGTGCGGCCTTCTCTGGCTTTGTCCAGGGCTTCTTGGACAACCTTTT CACTTTCAGTATCCAGAGCTGATGTAGCTTCATCCAACAGGAGGATTTGAGGTTGTCTG ATGAGGGCTCGGGCAATAGCAATCCTCTGTTTTTGACCTCCTGAGAGCTGAGTCCCCTT ATCTCCCACTCTTGTTTCATATTTGTGGGGTAACGTCTCGATGAAAGGATGTATGTTGGC AGCTTTGGCTGCACTCACAATTTCATCCTGTGATACAACCCGGCTGTTGTCTCCATAGG CAATATTCTCGGCAATGCTGCAGTCAAATAGGATAGGCTCCTGAGACACGATTCCGAGT TGAGCTCTGAGCCACTGGACATTGAGTTTCTTTGCTTCTTGACCATCGAGAAGCACTGT CCCCGCCAAGGGGTCGTAGAACCGCTCCAGGAGCTGGACCACCGTGCTCTTCCCACA GCCACTGCTGCCCACCAGGGCTAGTGTCTGGCCTTTCTTCACCTCCAGGCTCAGCCCC TGAAGCACTGGCACGTTTGCTCGGGTGGGATAGTTGAACACGACTTCATTAAATGTTAT ATTTCCTTCAAATTTATCAGGCTTCAGCCCCTCTTCACTGTAGCTGTCAATCAGAGGTTG TCTTTCAAACAGCATGAATAAGTGGGCTGCAGACAGCTTAGCTTTAGCATAGTCTGGAG CAAATGAACTGGCATGTCCTAGAGCCACTGCACCAAATACAATTGCAGAAAACACCAGA ATAACATCTCTGAAGCGCATATGTCCATTCACAATGAGATATGCACCAAATCGAAAACAA CCGGCATAGGAAAAATACATAAATGCTTGTGAGATACTAAAAGTAATTCCATAGATGTGT GCCTTCTGCACAGAATTCCTGTAAGGTCCATACAATTTTTCAACATACATTGATTCAAATT TTCTTTCCTGGGTCAAAGACACAACTGTCCTAATATTTTCTATTGCCTCTGTTGCAATCTT TCCAGCAGCTTCCAGTTCTTTTTTATCTCTTTTGGCATTTCCAGCCAACAATTTCATTTCA ACAATTCCTGACACAGCAATAATTGGAACAACTGCTAATAGCAATAGGGTTAACTGCCAA CCGTAGATAAATGATATGATAATACCAGTTCCAAGGTTAGCTATATTCTGTGCAATTAAA GCCAACCTGGTTCCTGTGGCTCCTTGGACTTGGGCAGCATCTGTGGCAAGTCTTGTAGA AAGTGCACCAGTACTGTTTTTATGGTCATCAAACCAGCTCATGTCCTGTCTTAGCATTGC TTTAAAAGCCATTGACCGCAGTCTTCTGGTGAGGATCTCGCCAGCTTTCCCAAACGTGA AACCCTGAAGGAAGAAAGTAAAAAAAGAAATAATTCCCAGAAATAAGAAAATCAAAGAGA ATATGTTGCACTTCTGCTGCTTCACTGCATCATCGCCTGGTCCAAAAATCGCTATGATCT CTGAGAATATGACTGAAAATGCCGGCTGAAGCCCCCCATTGGCAATGGCACATACTGTT CCCACGACAAAGTAGGGCCATTCTGTTTTATTCAGTTTCAGGACCTTCAGAAAGGACAC TGGTGGCACATTTGCTTCAAGTCCATCGGTTTCCACATCAAGGCTCTTCTGACACATTTG TGAATTTTTAAGGTTTTTCTGAGTAGAATGCCTAAATAGGCGAGATTTCCAGCCATTTGG GGCCATTCTAGTGGCAGCCTTTTCATCATTTAGTTCAAATTCTTCTGACTGGATCTGGCT TCCTGATGTCTGCATGTTGACAAGTTTGAAGTACACCCCTTCCTTCTTCATCAGTTCGCT GTGGCTTCCTTGCTCCACAATTACTCCATCCTCAAACCCAGCGATGACATCTGCATTTCG GACCGTAGACAGTCGGTGTGCTATCACAATGGTGGTCCGGCCTTCTCTGGCCTTATCCA GAGCTGCCTGTACCTCAGCTTCACTTTCTGTGTCCAATGCTGACGTGGCCTCATCCAGC AGAAGGATCTTGGGGTTGCGAACCAGGGCACGTGCAATGGCGATCCTCTGCTTCTGCC CACCACTCAGCTGGGCCCCTCTCTCTCCAACCAGGGTGTCAAATTTCTGTGGTAATTTC ATGATAAACTCATAGGCGTTGGCCTCTTTGACAGCTTTCTTTATCTCATCCATGGTTACA TTTCCACGGCCATAACAAATATTTTCAGCAATTGTGGTGGAAAACAGCACCGGCTCCTG ACTCACCACACCAATGATTTCCCTCAGATAGTTTACATTAAAGTTCCTAATATCCTGCCC ATCAATGTTAATTGTGCCCTCATCAGGGTCATAGAGCCTCTGTATCAGCTGGACCGTTG TGCTCTTCCCACAGCCACTACTTCCAACCAGGGCCACCGTCTGCCCACTCTGCACCTTC AGGTTGAGGCCCTTCAAGATCTTGACGTTAGCTCGAGAAGGGTAAGAAAAGTGAACATC ATTGAACTCCAAATTCCCTTTGATGCTGTCTGGTTTGTGTCCTCTCTCTGAAAAACTGTC AATTTTAGGATTATTATCAATAATATCAAAGATCACATATGCTGCTCCTCTTGCATTGGCA AAAGCATCAATACATGGGGCAGCCTGGCCAACACTGAAAGCTCCAATTAGGATTGAAAA AAAAACTGTCATTGCATTTCCAATAGTATATTCTTTTGATATGACTAGAGTGGATCCATAC CAGAAGGCCAGTGCATATGATGCATATATTAACAGGAAGGCAATACCCATGGAAATGTTTGCTGAAATAGCTTTTTTAATTCCAATCTCTTTGGCATTTTCTAAATGTTTCTGATACCTTT CCAGCTCTTTGTTCTGGCCCCCGAAAGCTATCACAGTCCTGATGGCCCCCAGAGCCTCT TCTGCCACGGCGCCTGCTTTTGCATAAGCAGCTAGTTCTTTGTCACTAAATGCCGAGAG TATCTTTGCCCAAACGGCTGCAGAGAGTCCTAGAATAGGGCTGATGGCCATTATCACAA GGGTGAGCTTCCATCCTCTGATGAATCCCACTATGAATCCTGCAAAAAACGTGGCTACT GCTTGAAAGAACATTCCAACCTTGTCACCAATTCCTTCACTGATTTTGGAGATGTCATCT GTTAGCCGCGTATTGAGTTCAGTGGTGTCGTTGATGTCAAACCATCCTATTTCCTGTCGT AGAATAGCATGAAAAAACTTCTGCCTAATTTTCCTGATCTGCCGACCAGCTGCCAAAGTC CAAAATGAAACTTGTATATAGGCAGCAACAAGAACTCCAGCACCCAATCCTGAGTAGTA ATATGCATATCTAGTCATTTCTTCTTCCAGAATTTTGCCTGGATTTAGCAGCGACAAGGA AAAGTTCACTGGAAAGGAGAAGTTTCCTGCAGTATCAACAAATTTGTCAGTCATCTCTCC AAATACTATCATCATGAGGGGGAGACCTGATCCGTGAGCTATGGCCATGATGGTACCCA GCGACATAAACAATTTATCCTGCCAATCGGAGTATCGAAACAATGTTAATACTCCAATCA TTTTCACTGTCTTCGTTTTTTTCCTTTTTTGTTTGCTGCTGATGCCCAGTTCAAAGTCGCC CTCCGCGCTCGTGGGGCGCCAGGCTGTTCCGTTCTTTGCCGCCTCAAGATCCATGGTG GCGGATCACCGGTCTCAGCGGGAGGAGAAACCACAGCCTCAGAACCAAGTACACCCTC TCCGGCGGCCCGGCGCACGAGTCGCGGGGCCCGGGGGCACTGGGTGGAGGTCCGGC CACTCCCGCCCCGCGCCCCACGTCCCGGGTCTCCCTTCGAGGCCAGACGCGCCCGGA CCTCTCTCGGCTCGAACCTCGCGCGTGTCTGGCAGGGCCTCTGGACGCGCGGGCGCT GCAGCAGAGGGGCCTGGACTTTGCTCGCCGCGGCCTCGCCCCCGCCCCCGCCCCCG CCCCCCACGCGCGTCCGGCCCGGGAAACGCCTACCGTTGCAGCCAGGGCGAGGGCC GCGGCCCGCGAGAGCAGCTCCCGCCGCCCGCCCAGCTCTCTCCCCAGGCTAGGCGC GGGCCAGGGGCGCAGCCTGCCGCTCTTGCCGCCGGGGCGCCCGGCTGGGGCCGCCG CCCACCCTGGCGACCCGCCAGAACCAGCCCGGTCGCAGGATAAGACCGAGATCAGGC TGGAGCCTCGGGTGCCGAGAAAGCCTCGCCTGATGGGCCACCCCCTCGAGGCCTCTC ACACTACCTAAACCACGCCAGGACAACCTCTGCTCCTCTCCACCGAAATTCCAAGGGGT CGAGTGGATGTTGGAGGTGGCATGGGCCCAGAGAGGTCTCTGACCTCTGCCCCAGCTC CAAGGTCAGCAGGCAGGGAGGGCTGTGTGTTTGCTGTTTGCTGCTTGCAATGTTTGCC CATTTTAGGGGATATGCTTGAATTCGAGCTCGGTACCTTTAAGACCAATGACTTACAAGG CAGCTGTAGATCTTAGCCACTTTTTAAAAGAAAAGGGGGGACTGGAAGGGCTAATTCAC TCCCAACGAAGACAAGATCTGCTTTTTGCTTGTACTGGGTCTCTCTGGTTAGACCAGATC TGAGCCTGGGAGCTCTCTGGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTT GCCTTGAGTGCTTCAAGTAGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGAT CCCTCAGACCCTTTTAGTCAGTGTGGAAAATCTCTAGCA SEQ ID NO: 16 (LTR-LTR of example construct comprising codon-optimised ABCB4) GGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGGCTAACTAGGGAACC CACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCCCGTCTG TTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAAAATCTCT AGCAGTGGCGCCCGAACAGGGACTTGAAAGCGAAAGGGAAACCAGAGGAGCTCTCTC GACGCAGGACTCGGCTTGCTGAAGCGCGCACGGCAAGAGGCGAGGGGCGGCGACTG GTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGAAGGAGAGAGATGGGTGCGAGA GCGTCAGTATTAAGCGGGGGAGAATTAGATCGCGATGGGAAAAAATTCGGTTAAGGCC AGGGGGAAAGAAAAAATATAAATTAAAACATATAGTATGGGCAAGCAGGGAGCTAGAAC GATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGTAGACAAATACTGGGA CAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATCATTATATAATACAGTA GCAACCCTCTATTGTGTGCATCAAAGGATAGAGATAAAAGACACCAAGGAAGCTTTAGA CAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCACAGCAAGCGGCCGCTGAT CTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAGTGAATTATATAAATATA AAGTAGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGCAAAGAGAAGAGTGGTG CAGAGAGAAAAAAGAGCAGTGGGAATAGGAGCTTTGTTCCTTGGGTTCTTGGGAGCAG CAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACAGGCCAGACAATTATT GTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTGAGGCGCAACAGCAT CTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAGAATCCTGGCTGTGG AAAGATACCTAAAGGATCAACAGCTCCTGGGGATTTGGGGTTGCTCTGGAAAACTCATTTGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTCTGGAACAGATTTG GAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTACACAAGCTTAATAC ACTCCTTAATTGAAGAATCGCAAAACCAGCAAGAAAAGAATGAACAAGAATTATTGGAAT TAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACATAACAAATTGGCTGTGGTATATAA AATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTTTTGCTGTACTTTC TATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGACCCACCTCCCAAC CCCGAGGGGACCCGACAGGCCCGAAGGAATAGAAGAAGAAGGTGGAGAGAGAGACAG AGACAGATCCATTCGATTAGTGAACGGATCTCGACGGTATCGGTTAACTTTTAAAAGAAA AGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGACATAATAGCAACAGAC ATACAAACTAAAGAATTACAAAAACAAATTACAAAAATTCAAAATTTTATCGATCACGAGA CTAGCCTCGAGGATATCCCATAGAGCCCACCGCATCCCCAGCATGCCTGCTATTGTCTT CCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCAGAATAGAATGACACCTA CTCAGACAATGCGATGCAATTTCCTCATTTTATTAGGAAAGGACAGTGGGAGTGGCACC TTCCAGGGTCAAGGAAGGCACGGGGGAGGGGCAAACAACAGATGGCTGGCAACTAGA AGGCACAGACGCGTGGGCCCCTGCAGTCGTACCGTGAGTAATAATGCGATCGTCGTAC CGTGAGTAATAATGCGCGGTTCGTACCGTGAGTAATAATGCGATCGTCGTACCGTGAGT AATAATGCGCTGCAGGTGTAGTGTTTCCTACTTTATGGAGTGATGTAGTGTTTCCTACTT TATGGACGGTTGTAGTGTTTCCTACTTTATGGAATCGTGTAGTGTTTCCTACTTTATGGA GGCTAGCCTAGGCGGGGAGGCGGCCCAAAGGGAGATCCGACTCGTCTGAGGGCGAAG GCGAAGACGCGGAAGAGGCCGCAGAGCCGGCAGCAGGCCGCGGGAAGGAAGGTCCG CTGGATTGAGGGCCGAAGGGACGTAGCAGAAGGACGTCCCGCGCAGAATCCAGGTGG CAACACAGGCGAGCAGCCATGGAAAGGACGTCAGCTTCCCCGACAACACCACGGAATT GTCAGTGCCCAACAGCCGAGCCCCTGTCCAGCAGCGGGCAAGGCAGGCGGCGATGAG TTCCGCCGTGGCAATAGGGAGGGGGAAAGCGAAAGTCCCGGAAAGGAGCTGACAGGT GGTGGCAATGCCCCAACCAGTGGGGGTTGCGTCAGCAAACACAGTGCACACCACGCCA CGTTGCCTGACAACGGGCCACAACTCCTCATAAAGAGACAGCAACCAGGATTTATACAA GGAGGAGAAAATGAAAGCCATACGGGAAGCAATAGCATGATACAAAGGCATTAAAGCA GCGTATCCACATAGCGTAAAAGGAGCAACATAGTTAAGAATACCAGTCAATCTTTCACAA ATTTTGTAATCCAGAGGTTGATTGTCGACTCACAGGTTCTGTGTGCCGGCCTGCACAGA CACCATGGAGAAGTAGATGCCCTTCTGGGCCAGCAGTTGCTGGTGGGTGCCGTGCTCT TTCACTCTGCCGTTCTGGAACACCACGATGAGGTCGGCATTCTGGATGGTGCTCAGCC GGTGAGCGATCACGATACAGGTTCTTCCCTCGCGGGCCTTATCCAGAGCCTCTTGCAC CACCTTCTCGCTCTCGGTGTCCAGAGCAGAGGTGGCCTCATCCAGGAGCAGAATCTGA GGCTGTCTGATCAGGGCGCGAGCGATAGCAATCCTCTGTTTCTGGCCTCCGCTCAGCT GTGTGCCCTTGTCGCCCACTCTTGTCTCGTATTTGTGGGGCAGTGTCTCGATGAAGGG GTGAATGTTGGCAGCCTTGGCAGCGGACACAATCTCGTCCTGGGACACGACTCTGCTG TTGTCGCCATAGGCAATATTCTCGGCAATGCTGCAGTCGAACAGGATGGGTTCTTGAGA CACGATTCCCAGCTGAGCCCTCAGCCACTGCACGTTCAGCTTTTTGGCTTCTTGGCCAT CCAGCAGGACGGTTCCAGCCAGAGGATCGTAGAACCGTTCCAGCAGCTGCACCACTGT AGACTTGCCACATCCGCTAGAGCCCACGAGAGCCAGAGTCTGGCCCTTCTTCACTTCCA GGCTCAGTCCCTGCAGAACTGGCACGTTGGCTCTGGTGGGGTAGTTGAACACCACCTC GTTAAAGGTGATGTTGCCCTCGAACTTATCTGGCTTCAGGCCTTCCTCGCTGTAGGAGT CGATCAGGGGCTGTCTCTCGAACAGCATGAACAGGTGTGCGGCAGACAGCTTGGCCTT GGCGTAGTCAGGGGCAAAGCTAGAGGCATGTCCCAGAGCCACGGCTCCAAACACGAT GGCGGAGAACACCAGGATCACGTCCCGGAATCTCATGTGGCCGTTCACGATCAGGTAG GCGCCGAATCTAAAGCAGCCGGCGTAGCTAAAGTACATGAAGGCCTGGCTGATGCTGA AGGTGATGCCGTAGATGTGGGCCTTCTGCACGCTGTTTCTGTAGGGGCCGTACAGCTT TTCCACGTACATAGACTCGAACTTCCGCTCTTGGGTCAGAGACACCACGGTCCGGATAT TCTCGATGGCCTCGGTGGCGATCTTGCCTGCGGCCTCAAGTTCTTTCTTGTCCCGCTTG GCGTTTCCGGCCAGCAACTTCATTTCCACGATGCCGGACACGGCAATGATAGGCACCA CGGCCAGCAGGAGCAGTGTCAGCTGCCATCCGTAGATGAAGCTAATGATGATGCCGGT GCCGAGATTGGCGATATTCTGGGCGATCAGGGCCAGTCTTGTGCCTGTAGCGCCTTGC ACCTGAGCAGCATCTGTGGCCAGTCTGGTAGACAGGGCGCCAGTGCTGTTCTTGTGGT CGTCGAACCAACTCATGTCCTGTCTCAGCATGGCTTTGAAGGCCATGGATCTCAGTCTC CGAGTCAGGATCTCCCCGGCCTTGCCGAAGGTGAAGCCTTGCAGGAAGAATGTAAAGAAGCTGATAATCCCCAGGAACAGGAAGATCAGGCTAAAGATATTACACTTCTGCTGCTTC ACGGCGTCGTCGCCAGGGCCAAAGATGGCGATAATCTCGCTAAAGATCACGGAGAAGG CGGGCTGCAGTCCACCATTAGCAATGGCACACACGGTGCCCACGACGAAGTAGGGCCA CTCGGTCTTGTTCAGTTTCAGCACTTTCAGGAAGGACACAGGAGGCACATTGGCTTCCA GTCCGTCTGTCTCGACGTCCAGGGACTTCTGGCACATCTGGCTGTTCTTCAGATTCTTC TGGGTGCTGTGTCTGAACAGCCGGGACTTCCAGCCATTAGGGGCCATCCTGGTGGCGG CCTTCTCGTCATTCAGCTCGAATTCCTCGCTCTGGATCTGAGAGCCGCTGGTCTGCATG TTGACCAGCTTGAAGTACACGCCTTCTTTCTTCATCAGCTCGCTGTGGGAGCCCTGTTC CACGATCACGCCATCTTCGAATCCGGCGATCACATCGGCATTCCGCACGGTGGACAGT CTGTGGGCAATCACGATGGTGGTTCTGCCCTCTCTAGCCTTGTCCAGGGCAGCCTGCA CTTCTGCCTCAGACTCTGTATCCAGGGCGCTTGTGGCTTCGTCCAGCAGCAGGATCTTA GGGTTCCGCACGAGTGCTCTGGCAATGGCGATTCTCTGCTTCTGTCCGCCAGACAGTT GGGCGCCTCTTTCTCCCACAAGGGTGTCGAATTTCTGGGGCAGCTTCATGATGAACTCG TAGGCGTTGGCCTCTTTCACGGCTTTCTTGATCTCGTCCATGGTCACGTTGCCTCTGCC GTAGCAGATGTTCTCGGCGATTGTGGTGCTGAACAGCACAGGCTCTTGGGACACCACT CCGATGATCTCGCGCAGGTAGTTCACGTTGAAGTTCCGGATGTCCTGGCCGTCGATGTT GATTGTGCCCTCGTCAGGGTCGTACAGTCTCTGGATCAGCTGCACGGTGGTGCTCTTG CCACAGCCAGAAGATCCCACGAGGGCCACTGTCTGTCCGCTCTGCACCTTCAGGTTCA GGCCCTTCAGGATCTTCACGTTGGCCCGGCTAGGGTAGCTGAAGTGCACGTCGTTGAA TTCCAGGTTGCCCTTGATGCTATCAGGCTTGTGGCCCCGCTCGCTGAAGCTGTCGATCT TGGGGTTGTTGTCGATGATATCGAAGATCACGTATGCGGCGCCTCTGGCATTAGCAAAG GCGTCGATACAAGGAGCGGCCTGTCCGACGCTAAAGGCGCCGATCAGGATGGAGAAG AACACGGTCATGGCGTTGCCGATGGTGTACTCTTTGCTGATGACCAGGGTAGAGCCGT ACCAAAAGGCCAGGGCGTAGCTGGCGTAGATCAGCAGGAAGGCGATTCCCATAGAGAT GTTGGCGCTGATGGCCTTCTTGATGCCAATCTCTTTGGCGTTCTCGAGGTGCTTCTGGT ATCTTTCCAGTTCTTTGTTCTGGCCGCCAAAGGCGATCACGGTTCTGATGGCTCCCAGG GCTTCTTCAGCAACGGCGCCAGCTTTAGCGTAAGCGGCCAGCTCTTTGTCGCTGAAGG CGCTCAGAATCTTGGCCCACACAGCAGCAGACAGTCCCAGGATGGGAGAGATGGCCAT GATCACGAGGGTCAGCTTCCAGCCTCTGATGAAGCCCACGATAAAGCCGGCGAAGAAG GTGGCCACGGCCTGGAAGAACATTCCCACTTTGTCGCCGATGCCTTCGCTGATCTTGCT GATGTCGTCGGTCAGTCTGGTGTTCAGCTCGGTGGTGTCGTTGATGTCGAACCATCCG ATCTCTTGCCGCAGGATGGCGTGGAAGAACTTCTGCCGGATCTTTCTGATCTGCCGGC CAGCGGCCAGTGTCCAAAAGGACACCTGGATATAGGCGGCCACCAGAACGCCAGCTCC AAGGCCGCTGTAGTAGTAGGCGTATCTGGTCATTTCCTCTTCCAGGATCTTGCCGGGGT TCAGCAGGCTCAGGCTGAAATTCACGGGGAAGCTGAAGTTGCCGGCGGTGTCCACGAA CTTGTCGGTCATCTCGCCGAACACGATCATCATCAGGGGCAGTCCAGATCCGTGGGCG ATAGCCATGATGGTGCCCAGGCTCATGAACAGCTTGTCCTGCCAGTCGCTGTACCGGA ACAGGGTCAGCACGCCGATCATCTTCACGGTCTTGGTCTTTTTCCGCTTCTGCTTGCTG CTGATGCCCAGCTCAAAATCGCCCTCGGCAGATGTAGGCCTCCAAGCGGTGCCATTCT TGGCGGCTTCCAGATCCATGGTGGCGGATCACCGGTCTCAGCGGGAGGAGAAACCAC AGCCTCAGAACCAAGTACACCCTCTCCGGCGGCCCGGCGCACGAGTCGCGGGGCCCG GGGGCACTGGGTGGAGGTCCGGCCACTCCCGCCCCGCGCCCCACGTCCCGGGTCTC CCTTCGAGGCCAGACGCGCCCGGACCTCTCTCGGCTCGAACCTCGCGCGTGTCTGGC AGGGCCTCTGGACGCGCGGGCGCTGCAGCAGAGGGGCCTGGACTTTGCTCGCCGCG GCCTCGCCCCCGCCCCCGCCCCCGCCCCCCACGCGCGTCCGGCCCGGGAAACGCCT ACCGTTGCAGCCAGGGCGAGGGCCGCGGCCCGCGAGAGCAGCTCCCGCCGCCCGCC CAGCTCTCTCCCCAGGCTAGGCGCGGGCCAGGGGCGCAGCCTGCCGCTCTTGCCGCC GGGGCGCCCGGCTGGGGCCGCCGCCCACCCTGGCGACCCGCCAGAACCAGCCCGGT CGCAGGATAAGACCGAGATCAGGCTGGAGCCTCGGGTGCCGAGAAAGCCTCGCCTGA TGGGCCACCCCCTCGAGGCCTCTCACACTACCTAAACCACGCCAGGACAACCTCTGCT CCTCTCCACCGAAATTCCAAGGGGTCGAGTGGATGTTGGAGGTGGCATGGGCCCAGAG AGGTCTCTGACCTCTGCCCCAGCTCCAAGGTCAGCAGGCAGGGAGGGCTGTGTGTTTG CTGTTTGCTGCTTGCAATGTTTGCCCATTTTAGGGGATATGCTTGAATTCGAGCTCGGTA CCTTTAAGACCAATGACTTACAAGGCAGCTGTAGATCTTAGCCACTTTTTAAAAGAAAAG GGGGGACTGGAAGGGCTAATTCACTCCCAACGAAGACAAGATCTGCTTTTTGCTTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGGCTAACTAGGGAAC CCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCCCGTCT GTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAAAATCTC TAGCA SEQ ID NO: 17 - reverse complement of woodchuck hepatitis virus posttranscriptional regulatory element (WPRE) gcggggaggcggcccaaagggagatccgactcgtctgagggcgaaggcgaagacgcggaagaggccgcagagccggca gcaggccgcgggaaggaaggtccgctggattgagggccgaagggacgtagcagaaggacgtcccgcgcagaatccaggtg gcaacacaggcgagcagccatggaaaggacgtcagcttccccgacaacaccacggaattgtcagtgcccaacagccgagc ccctgtccagcagcgggcaaggcaggcggcgatgagttccgccgtggcaatagggagggggaaagcgaaagtcccggaaa ggagctgacaggtggtggcaatgccccaaccagtgggggttgcgtcagcaaacacagtgcacaccacgccacgttgcctgac aacgggccacaactcctcataaagagacagcaaccaggatttatacaaggaggagaaaatgaaagccatacgggaagcaa tagcatgatacaaaggcattaaagcagcgtatccacatagcgtaaaaggagcaacatagttaagaataccagtcaatctttcac aaattttgtaatccagaggttgatt

[0225] SEQ ID NO: 18 - 5' LTR (truncated)- truncated 5' long terminal repeat (LTR) from HIV-1 gggtctctctggttagaccagatctgagcctgggagctctctggctaactagggaacccactgcttaagcctcaataaagcttgcctt gagtgcttcaagtagtgtgtgcccgtctgttgtgtgactctggtaactagagatccctcagacccttttagtcagtgtggaaaatctcta gca

[0226] SEQ ID NO: 19 - HIV-1

[0227]

[0228] packaging signal of human immunodeficiency virus type 1 ctctctcgacgcaggactcggcttgctgaagcgcgcacggcaagaggcgaggggcggcgactggtgagtacgccaaaaatttt gactagcggaggctagaaggagagagatgggtgcgagagcgtc

[0229] SEQ ID NO: 20 - cPPT / CTS_ central polypurine tract and central termination sequence of HIV-1 ttttaaaagaaaaggggggattggggggtacagtgcaggggaaagaatagtagacataatagcaacagacatacaaactaa agaattacaaaaacaaattacaaaaattcaaaatttt

[0230] SEQ ID NO: 21 - reverse complement of bGH poly(A) signal_ bovine growth hormone polyadenylation signal CCATAGAGCCCACCGCATCCCCAGCATGCCTGCTATTGTCTTCCCAATCCTCCCCCTTG CTGTCCTGCCCCACCCCACCCCCCAGAATAGAATGACACCTACTCAGACAATGCGATGC AATTTCCTCATTTTATTAGGAAAGGACAGTGGGAGTGGCACCTTCCAGGGTCAAGGAAG GCACGGGGGAGGGGCAAACAACAGATGGCTGGCAACTAGAAGGCACAG SEQ ID NO: 23 - Portion of naturally occurring ABCB4 human promoter (Comparative example) GGGTGGCCCATCAGGCGAGGCTTTCTCGGCACCCGAGGCTCCAGCCTGATCTCGGTCT TATCCTGCGACCGGGCTGGTTCTGGCGGGTCGCCAGGGTGGGCGGCGGCCCCAGCC GGGCGCCCCGGCGGCAAGAGCGGCAGGCTGCGCCCCTGGCCCGCGCCTAGCCTGGG GAGAGAGCTGGGCGGGCGGCGGGAGCTGCTCTCGCGGGCCGCGGCCCTCGCCCTGG CTGCAACGGTAGGCGTTTCCCGGGCCGGACGCGCGTGGGGGGCGGGGGCGGGGGC GGGGGCGAGGCCGCGGCGAGCAAAGTCCAGGCCCCTCTGCTGCAGCGCCCGCGCGT CCAGAGGCCCTGCCAGACACGCGCGAGGTTCGAGGTGAGAGAGGTCCGGGCGCGTCT GGCCTCGAAGGGAGACCCGGGACGTGGGGCGCGGGGCGGGAGTGGCCGGACCTCCA CCCAGTGCCCCCGGGCCCCGCGACTCGTGCGCCGGGCCGCCGGAGAGGGTGTACTT GGTTCTGAGGCTGTGGTTTCTCCTCAGGCTGAGA SEQ ID NO: 24 - Reverse complement of portion of naturally occurring ABCB4 human promoter (Comparative example) TCTCAGCCTGAGGAGAAACCACAGCCTCAGAACCAAGTACACCCTCTCCGGCGGCCCG GCGCACGAGTCGCGGGGCCCGGGGGCACTGGGTGGAGGTCCGGCCACTCCCGCCCC GCGCCCCACGTCCCGGGTCTCCCTTCGAGGCCAGACGCGCCCGGACCTCTCTCACCTCGAACCTCGCGCGTGTCTGGCAGGGCCTCTGGACGCGCGGGCGCTGCAGCAGAGGG GCCTGGACTTTGCTCGCCGCGGCCTCGCCCCCGCCCCCGCCCCCGCCCCCCACGCGC GTCCGGCCCGGGAAACGCCTACCGTTGCAGCCAGGGCGAGGGCCGCGGCCCGCGAG AGCAGCTCCCGCCGCCCGCCCAGCTCTCTCCCCAGGCTAGGCGCGGGCCAGGGGCG CAGCCTGCCGCTCTTGCCGCCGGGGCGCCCGGCTGGGGCCGCCGCCCACCCTGGCG ACCCGCCAGAACCAGCCCGGTCGCAGGATAAGACCGAGATCAGGCTGGAGCCTCGGG TGCCGAGAAAGCCTCGCCTGATGGGCCACCC SEQ ID NO: 25 - LPa promoter + coABCB4 transgene sequence TCATAAGTTCTGTGTCCCAGCCTGGACACTGACCATTGAAAAATAGATGCCTTTCTGTGC CAGCAGCTGCTGATGCGTGCCATGCTCCTTGACTCTCCCATTCTGAAACACCACTATTA AGTCTGCATTCTGGATGGTGGACAGGCGGTGAGCAATCACAATGCAGGTGCGGCCTTC TCTGGCTTTGTCCAGGGCTTCTTGGACAACCTTTTCACTTTCAGTATCCAGAGCTGATGT AGCTTCATCCAACAGGAGGATTTGAGGTTGTCTGATGAGGGCTCGGGCAATAGCAATCC TCTGTTTTTGACCTCCTGAGAGCTGAGTCCCCTTATCTCCCACTCTTGTTTCATATTTGT GGGGTAACGTCTCGATGAAAGGATGTATGTTGGCAGCTTTGGCTGCACTCACAATTTCA TCCTGTGATACAACCCGGCTGTTGTCTCCATAGGCAATATTCTCGGCAATGCTGCAGTC AAATAGGATAGGCTCCTGAGACACGATTCCGAGTTGAGCTCTGAGCCACTGGACATTGA GTTTCTTTGCTTCTTGACCATCGAGAAGCACTGTCCCCGCCAAGGGGTCGTAGAACCGC TCCAGGAGCTGGACCACCGTGCTCTTCCCACAGCCACTGCTGCCCACCAGGGCTAGTG TCTGGCCTTTCTTCACCTCCAGGCTCAGCCCCTGAAGCACTGGCACGTTTGCTCGGGT GGGATAGTTGAACACGACTTCATTAAATGTTATATTTCCTTCAAATTTATCAGGCTTCAGC CCCTCTTCACTGTAGCTGTCAATCAGAGGTTGTCTTTCAAACAGCATGAATAAGTGGGCT GCAGACAGCTTAGCTTTAGCATAGTCTGGAGCAAATGAACTGGCATGTCCTAGAGCCAC TGCACCAAATACAATTGCAGAAAACACCAGAATAACATCTCTGAAGCGCATATGTCCATT CACAATGAGATATGCACCAAATCGAAAACAACCGGCATAGGAAAAATACATAAATGCTT GTGAGATACTAAAAGTAATTCCATAGATGTGTGCCTTCTGCACAGAATTCCTGTAAGGTC CATACAATTTTTCAACATACATTGATTCAAATTTTCTTTCCTGGGTCAAAGACACAACTGT CCTAATATTTTCTATTGCCTCTGTTGCAATCTTTCCAGCAGCTTCCAGTTCTTTTTTATCT CTTTTGGCATTTCCAGCCAACAATTTCATTTCAACAATTCCTGACACAGCAATAATTGGAA CAACTGCTAATAGCAATAGGGTTAACTGCCAACCGTAGATAAATGATATGATAATACCAG TTCCAAGGTTAGCTATATTCTGTGCAATTAAAGCCAACCTGGTTCCTGTGGCTCCTTGGA CTTGGGCAGCATCTGTGGCAAGTCTTGTAGAAAGTGCACCAGTACTGTTTTTATGGTCA TCAAACCAGCTCATGTCCTGTCTTAGCATTGCTTTAAAAGCCATTGACCGCAGTCTTCTG GTGAGGATCTCGCCAGCTTTCCCAAACGTGAAACCCTGAAGGAAGAAAGTAAAAAAAGA AATAATTCCCAGAAATAAGAAAATCAAAGAGAATATGTTGCACTTCTGCTGCTTCACTGC ATCATCGCCTGGTCCAAAAATCGCTATGATCTCTGAGAATATGACTGAAAATGCCGGCT GAAGCCCCCCATTGGCAATGGCACATACTGTTCCCACGACAAAGTAGGGCCATTCTGTT TTATTCAGTTTCAGGACCTTCAGAAAGGACACTGGTGGCACATTTGCTTCAAGTCCATCG GTTTCCACATCAAGGCTCTTCTGACACATTTGTGAATTTTTAAGGTTTTTCTGAGTAGAAT GCCTAAATAGGCGAGATTTCCAGCCATTTGGGGCCATTCTAGTGGCAGCCTTTTCATCA TTTAGTTCAAATTCTTCTGACTGGATCTGGCTTCCTGATGTCTGCATGTTGACAAGTTTG AAGTACACCCCTTCCTTCTTCATCAGTTCGCTGTGGCTTCCTTGCTCCACAATTACTCCA TCCTCAAACCCAGCGATGACATCTGCATTTCGGACCGTAGACAGTCGGTGTGCTATCAC AATGGTGGTCCGGCCTTCTCTGGCCTTATCCAGAGCTGCCTGTACCTCAGCTTCACTTT CTGTGTCCAATGCTGACGTGGCCTCATCCAGCAGAAGGATCTTGGGGTTGCGAACCAG GGCACGTGCAATGGCGATCCTCTGCTTCTGCCCACCACTCAGCTGGGCCCCTCTCTCT CCAACCAGGGTGTCAAATTTCTGTGGTAATTTCATGATAAACTCATAGGCGTTGGCCTCT TTGACAGCTTTCTTTATCTCATCCATGGTTACATTTCCACGGCCATAACAAATATTTTCAG CAATTGTGGTGGAAAACAGCACCGGCTCCTGACTCACCACACCAATGATTTCCCTCAGA TAGTTTACATTAAAGTTCCTAATATCCTGCCCATCAATGTTAATTGTGCCCTCATCAGGGT CATAGAGCCTCTGTATCAGCTGGACCGTTGTGCTCTTCCCACAGCCACTACTTCCAACC AGGGCCACCGTCTGCCCACTCTGCACCTTCAGGTTGAGGCCCTTCAAGATCTTGACGTT AGCTCGAGAAGGGTAAGAAAAGTGAACATCATTGAACTCCAAATTCCCTTTGATGCTGT CTGGTTTGTGTCCTCTCTCTGAAAAACTGTCAATTTTAGGATTATTATCAATAATATCAAAGATCACATATGCTGCTCCTCTTGCATTGGCAAAAGCATCAATACATGGGGCAGCCTGGC CAACACTGAAAGCTCCAATTAGGATTGAAAAAAAAACTGTCATTGCATTTCCAATAGTAT ATTCTTTTGATATGACTAGAGTGGATCCATACCAGAAGGCCAGTGCATATGATGCATATA TTAACAGGAAGGCAATACCCATGGAAATGTTTGCTGAAATAGCTTTTTTAATTCCAATCT CTTTGGCATTTTCTAAATGTTTCTGATACCTTTCCAGCTCTTTGTTCTGGCCCCCGAAAG CTATCACAGTCCTGATGGCCCCCAGAGCCTCTTCTGCCACGGCGCCTGCTTTTGCATAA GCAGCTAGTTCTTTGTCACTAAATGCCGAGAGTATCTTTGCCCAAACGGCTGCAGAGAG TCCTAGAATAGGGCTGATGGCCATTATCACAAGGGTGAGCTTCCATCCTCTGATGAATC CCACTATGAATCCTGCAAAAAACGTGGCTACTGCTTGAAAGAACATTCCAACCTTGTCAC CAATTCCTTCACTGATTTTGGAGATGTCATCTGTTAGCCGCGTATTGAGTTCAGTGGTGT CGTTGATGTCAAACCATCCTATTTCCTGTCGTAGAATAGCATGAAAAAACTTCTGCCTAA TTTTCCTGATCTGCCGACCAGCTGCCAAAGTCCAAAATGAAACTTGTATATAGGCAGCAA CAAGAACTCCAGCACCCAATCCTGAGTAGTAATATGCATATCTAGTCATTTCTTCTTCCA GAATTTTGCCTGGATTTAGCAGCGACAAGGAAAAGTTCACTGGAAAGGAGAAGTTTCCT GCAGTATCAACAAATTTGTCAGTCATCTCTCCAAATACTATCATCATGAGGGGGAGACCT GATCCGTGAGCTATGGCCATGATGGTACCCAGCGACATAAACAATTTATCCTGCCAATC GGAGTATCGAAACAATGTTAATACTCCAATCATTTTCACTGTCTTCGTTTTTTTCCTTTTTT GTTTGCTGCTGATGCCCAGTTCAAAGTCGCCCTCCGCGCTCGTGGGGCGCCAGGCTGT TCCGTTCTTTGCCGCCTCAAGATCCATGGTGGCGGATCACCGGTCTCAGCGGGAGGAG AAACCACAGCCTCAGAACCAAGTACACCCTCTCCGGCGGCCCGGCGCACGAGTCGCG GGGCCCGGGGGCACTGGGTGGAGGTCCGGCCACTCCCGCCCCGCGCCCCACGTCCC GGGTCTCCCTTCGAGGCCAGACGCGCCCGGACCTCTCTCGGCTCGAACCTCGCGCGT GTCTGGCAGGGCCTCTGGACGCGCGGGCGCTGCAGCAGAGGGGCCTGGACTTTGCTC GCCGCGGCCTCGCCCCCGCCCCCGCCCCCGCCCCCCACGCGCGTCCGGCCCGGGAA ACGCCTACCGTTGCAGCCAGGGCGAGGGCCGCGGCCCGCGAGAGCAGCTCCCGCCG CCCGCCCAGCTCTCTCCCCAGGCTAGGCGCGGGCCAGGGGCGCAGCCTGCCGCTCTT GCCGCCGGGGCGCCCGGCTGGGGCCGCCGCCCACCCTGGCGACCCGCCAGAACCAG CCCGGTCGCAGGATAAGACCGAGATCAGGCTGGAGCCTCGGGTGCCGAGAAAGCCTC GCCTGATGGGCCACCC SEQ ID NO: 26 - HCR + Lpa Promoter + wild-type ABCB4 transgene sequence TCATAAGTTCTGTGTCCCAGCCTGGACACTGACCATTGAAAAATAGATGCCTTTCTGTGC CAGCAGCTGCTGATGCGTGCCATGCTCCTTGACTCTCCCATTCTGAAACACCACTATTA AGTCTGCATTCTGGATGGTGGACAGGCGGTGAGCAATCACAATGCAGGTGCGGCCTTC TCTGGCTTTGTCCAGGGCTTCTTGGACAACCTTTTCACTTTCAGTATCCAGAGCTGATGT AGCTTCATCCAACAGGAGGATTTGAGGTTGTCTGATGAGGGCTCGGGCAATAGCAATCC TCTGTTTTTGACCTCCTGAGAGCTGAGTCCCCTTATCTCCCACTCTTGTTTCATATTTGT GGGGTAACGTCTCGATGAAAGGATGTATGTTGGCAGCTTTGGCTGCACTCACAATTTCA TCCTGTGATACAACCCGGCTGTTGTCTCCATAGGCAATATTCTCGGCAATGCTGCAGTC AAATAGGATAGGCTCCTGAGACACGATTCCGAGTTGAGCTCTGAGCCACTGGACATTGA GTTTCTTTGCTTCTTGACCATCGAGAAGCACTGTCCCCGCCAAGGGGTCGTAGAACCGC TCCAGGAGCTGGACCACCGTGCTCTTCCCACAGCCACTGCTGCCCACCAGGGCTAGTG TCTGGCCTTTCTTCACCTCCAGGCTCAGCCCCTGAAGCACTGGCACGTTTGCTCGGGT GGGATAGTTGAACACGACTTCATTAAATGTTATATTTCCTTCAAATTTATCAGGCTTCAGC CCCTCTTCACTGTAGCTGTCAATCAGAGGTTGTCTTTCAAACAGCATGAATAAGTGGGCT GCAGACAGCTTAGCTTTAGCATAGTCTGGAGCAAATGAACTGGCATGTCCTAGAGCCAC TGCACCAAATACAATTGCAGAAAACACCAGAATAACATCTCTGAAGCGCATATGTCCATT CACAATGAGATATGCACCAAATCGAAAACAACCGGCATAGGAAAAATACATAAATGCTT GTGAGATACTAAAAGTAATTCCATAGATGTGTGCCTTCTGCACAGAATTCCTGTAAGGTC CATACAATTTTTCAACATACATTGATTCAAATTTTCTTTCCTGGGTCAAAGACACAACTGT CCTAATATTTTCTATTGCCTCTGTTGCAATCTTTCCAGCAGCTTCCAGTTCTTTTTTATCT CTTTTGGCATTTCCAGCCAACAATTTCATTTCAACAATTCCTGACACAGCAATAATTGGAA CAACTGCTAATAGCAATAGGGTTAACTGCCAACCGTAGATAAATGATATGATAATACCAG TTCCAAGGTTAGCTATATTCTGTGCAATTAAAGCCAACCTGGTTCCTGTGGCTCCTTGGA CTTGGGCAGCATCTGTGGCAAGTCTTGTAGAAAGTGCACCAGTACTGTTTTTATGGTCATCAAACCAGCTCATGTCCTGTCTTAGCATTGCTTTAAAAGCCATTGACCGCAGTCTTCTG GTGAGGATCTCGCCAGCTTTCCCAAACGTGAAACCCTGAAGGAAGAAAGTAAAAAAAGA AATAATTCCCAGAAATAAGGGACACTGGTGGCACATTTGCTTCAAGTCCATCGGTTTCCA CATCAAGGCTCTTCTGACACATTTGTGAATTTTTAAGGTTTTTCTGAGTAGAATGCCTAAA TAGGCGAGATTTCCAGCCATTTGGGGCCATTCTAGTGGCAGCCTTTTCATCATTTAGTTC AAATTCTTCTGACTGGATCTGGCTTCCTGATGTCTGCATGTTGACAAGTTTGAAGTACAC CCCTTCCTTCTTCATCAGTTCGCTGTGGCTTCCTTGCTCCACAATTACTCCATCCTCAAA CCCAGCGATGACATCTGCATTTCGGACCGTAGACAGTCGGTGTGCTATCACAATGGTG GTCCGGCCTTCTCTGGCCTTATCCAGAGCTGCCTGTACCTCAGCTTCACTTTCTGTGTC CAATGCTGACGTGGCCTCATCCAGCAGAAGGATCTTGGGGTTGCGAACCAGGGCACGT GCAATGGCGATCCTCTGCTTCTGCCCACCACTCAGCTGGGCCCCTCTCTCTCCAACCA GGGTGTCAAATTTCTGTGGTAATTTCATGATAAACTCATAGGCGTTGGCCTCTTTGACAG CTTTCTTTATCTCATCCATGGTTACATTTCCACGGCCATAACAAATATTTTCAGCAATTGT GGTGGAAAACAGCACCGGCTCCTGACTCACCACACCAATGATTTCCCTCAGATAGTTTA CATTAAAGTTCCTAATATCCTGCCCATCAATGTTAATTGTGCCCTCATCAGGGTCATAGA GCCTCTGTATCAGCTGGACCGTTGTGCTCTTCCCACAGCCACTACTTCCAACCAGGGCC ACCGTCTGCCCACTCTGCACCTTCAGGTTGAGGCCCTTCAAGATCTTGACGTTAGCTCG AGAAGGGTAAGAAAAGTGAACATCATTGAACTCCAAATTCCCTTTGATGCTGTCTGGTTT GTGTCCTCTCTCTGAAAAACTGTCAATTTTAGGATTATTATCAATAATATCAAAGATCACA TATGCTGCTCCTCTTGCATTGGCAAAAGCATCAATACATGGGGCAGCCTGGCCAACACT GAAAGCTCCAATTAGGATTGAAAAAAAAACTGTCATTGCATTTCCAATAGTATATTCTTTT GATATGACTAGAGTGGATCCATACCAGAAGGCCAGTGCATATGATGCATATATTAACAG GAAGGCAATACCCATGGAAATGTTTGCTGAAATAGCTTTTTTAATTCCAATCTCTTTGGC ATTTTCTAAATGTTTCTGATACCTTTCCAGCTCTTTGTTCTGGCCCCCGAAAGCTATCACA GTCCTGATGGCCCCCAGAGCCTCTTCTGCCACGGCGCCTGCTTTTGCATAAGCAGCTA GTTCTTTGTCACTAAATGCCGAGAGTATCTTTGCCCAAACGGCTGCAGAGAGTCCTAGA ATAGGGCTGATGGCCATTATCACAAGGGTGAGCTTCCATCCTCTGATGAATCCCACTAT GAATCCTGCAAAAAACGTGGCTACTGCTTGAAAGAACATTCCAACCTTGTCACCAATTCC TTCACTGATTTTGGAGATGTCATCTGTTAGCCGCGTATTGAGTTCAGTGGTGTCGTTGAT GTCAAACCATCCTATTTCCTGTCGTAGAATAGCATGAAAAAACTTCTGCCTAATTTTCCT GATCTGCCGACCAGCTGCCAAAGTCCAAAATGAAACTTGTATATAGGCAGCAACAAGAA CTCCAGCACCCAATCCTGAGTAGTAATATGCATATCTAGTCATTTCTTCTTCCAGAATTTT GCCTGGATTTAGCAGCGACAAGGAAAAGTTCACTGGAAAGGAGAAGTTTCCTGCAGTAT CAACAAATTTGTCAGTCATCTCTCCAAATACTATCATCATGAGGGGGAGACCTGATCCGT GAGCTATGGCCATGATGGTACCCAGCGACATAAACAATTTATCCTGCCAATCGGAGTAT CGAAACAATGTTAATACTCCAATCATTTTCACTGTCTTCGTTTTTTTCCTTTTTTGTTTGCT GCTGATGCCCAGTTCAAAGTCGCCCTCCGCGCTCGTGGGGCGCCAGGCTGTTCCGTTC TTTGCCGCCTCAAGATCCATGGTGGCGGATCACCGGTCTCAGCGGGAGGAGAAACCAC AGCCTCAGAACCAAGTACACCCTCTCCGGCGGCCCGGCGCACGAGTCGCGGGGCCCG GGGGCACTGGGTGGAGGTCCGGCCACTCCCGCCCCGCGCCCCACGTCCCGGGTCTC CCTTCGAGGCCAGACGCGCCCGGACCTCTCTCGGCTCGAACCTCGCGCGTGTCTGGC AGGGCCTCTGGACGCGCGGGCGCTGCAGCAGAGGGGCCTGGACTTTGCTCGCCGCG GCCTCGCCCCCGCCCCCGCCCCCGCCCCCCACGCGCGTCCGGCCCGGGAAACGCCT ACCGTTGCAGCCAGGGCGAGGGCCGCGGCCCGCGAGAGCAGCTCCCGCCGCCCGCC CAGCTCTCTCCCCAGGCTAGGCGCGGGCCAGGGGCGCAGCCTGCCGCTCTTGCCGCC GGGGCGCCCGGCTGGGGCCGCCGCCCACCCTGGCGACCCGCCAGAACCAGCCCGGT CGCAGGATAAGACCGAGATCAGGCTGGAGCCTCGGGTGCCGAGAAAGCCTCGCCTGA TGGGCCACCCCCTCGAGGCCTCTCACACTACCTAAACCACGCCAGGACAACCTCTGCT CCTCTCCACCGAAATTCCAAGGGGTCGAGTGGATGTTGGAGGTGGCATGGGCCCAGAG AGGTCTCTGACCTCTGCCCCAGCTCCAAGGTCAGCAGGCAGGGAGGGCTGTGTGTTTG CTGTTTGCTGCTTGCAATGTTTGCCCATTTTAGGG SEQ ID NO: 27 - Proposed FXR binding region

[0231] AggtccgggcgcgtctggcctcgaagggagacccgggacgtggggcgcggggcgggagtggccggaccSEQ ID NO: 40 - hAAT promoter AATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGG GCAGCGTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGCCCCTGTTTGCTCCTC CGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGG ATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCAC CACTGACCTGGGACAGTGAATC SEQ ID NO: 41 - HCR forward CCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTG CTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTCTGGGCCCATGCCACCTCCAAC ATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTT TAGGTAGTGTGAGAGGGG SEQ ID NO: 42 - Comparative LP1 promoter CCCTAAAATGGGCAAACATTGCAAGCAGCAAACAGCAAACACACAGCCCTCCCTGCCTG CTGACCTTGGAGCTGGGGCAGAGGTCAGAGACCTCTCTGGGCCCATGCCACCTCCAAC ATCCACTCGACCCCTTGGAATTTCGGTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTT TAGGTAGTGTGAGAGGGGAATGACTCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTG CCCAGGCAAAGCGTCCGGGCAGCGTAGGCGGGCGACTCAGATCCCAGCCAGTGGACT TAGCCCCTGTTTGCTCCTCCGATAACTGGGGTGACCTTGGTTAATATTCACCAGCAGCC TCCCCCGTTGCCCCTCTGGATCCACTGCTTAAATACGGACGAGGACAGGGCCCTGTCT CCTCAGCTTCAGGCACCACCACTGACCTGGGACAGTGAATC SEQ ID NO: 43 - Wild type human ABCB4 coding sequence (CDS) ATGGATCTTGAGGCGGCAAAGAACGGAACAGCCTGGCGCCCCACGAGCGCGGAGGGC GACTTTGAACTGGGCATCAGCAGCAAACAAAAAAGGAAAAAAACGAAGACAGTGAAAAT GATTGGAGTATTAACATTGTTTCGATACTCCGATTGGCAGGATAAATTGTTTATGTCGCT GGGTACCATCATGGCCATAGCTCACGGATCAGGTCTCCCCCTCATGATGATAGTATTTG GAGAGATGACTGACAAATTTGTTGATACTGCAGGAAACTTCTCCTTTCCAGTGAACTTTT CCTTGTCGCTGCTAAATCCAGGCAAAATTCTGGAAGAAGAAATGACTAGATATGCATATT ACTACTCAGGATTGGGTGCTGGAGTTCTTGTTGCTGCCTATATACAAGTTTCATTTTGGA CTTTGGCAGCTGGTCGACAGATCAGGAAAATTAGGCAGAAGTTTTTTCATGCTATTCTAC GACAGGAAATAGGATGGTTTGACATCAACGACACCACTGAACTCAATACGCGGCTAACA GATGACATCTCCAAAATCAGTGAAGGAATTGGTGACAAGGTTGGAATGTTCTTTCAAGC AGTAGCCACGTTTTTTGCAGGATTCATAGTGGGATTCATCAGAGGATGGAAGCTCACCC TTGTGATAATGGCCATCAGCCCTATTCTAGGACTCTCTGCAGCCGTTTGGGCAAAGATA CTCTCGGCATTTAGTGACAAAGAACTAGCTGCTTATGCAAAAGCAGGCGCCGTGGCAGA AGAGGCTCTGGGGGCCATCAGGACTGTGATAGCTTTCGGGGGCCAGAACAAAGAGCTG GAAAGGTATCAGAAACATTTAGAAAATGCCAAAGAGATTGGAATTAAAAAAGCTATTTCA GCAAACATTTCCATGGGTATTGCCTTCCTGTTAATATATGCATCATATGCACTGGCCTTC TGGTATGGATCCACTCTAGTCATATCAAAAGAATATACTATTGGAAATGCAATGACAGTT TTTTTTTCAATCCTAATTGGAGCTTTCAGTGTTGGCCAGGCTGCCCCATGTATTGATGCT TTTGCCAATGCAAGAGGAGCAGCATATGTGATCTTTGATATTATTGATAATAATCCTAAAA TTGACAGTTTTTCAGAGAGAGGACACAAACCAGACAGCATCAAAGGGAATTTGGAGTTC AATGATGTTCACTTTTCTTACCCTTCTCGAGCTAACGTCAAGATCTTGAAGGGCCTCAAC CTGAAGGTGCAGAGTGGGCAGACGGTGGCCCTGGTTGGAAGTAGTGGCTGTGGGAAG AGCACAACGGTCCAGCTGATACAGAGGCTCTATGACCCTGATGAGGGCACAATTAACAT TGATGGGCAGGATATTAGGAACTTTAATGTAAACTATCTGAGGGAAATCATTGGTGTGGT GAGTCAGGAGCCGGTGCTGTTTTCCACCACAATTGCTGAAAATATTTGTTATGGCCGTG GAAATGTAACCATGGATGAGATAAAGAAAGCTGTCAAAGAGGCCAACGCCTATGAGTTT ATCATGAAATTACCACAGAAATTTGACACCCTGGTTGGAGAGAGAGGGGCCCAGCTGA GTGGTGGGCAGAAGCAGAGGATCGCCATTGCACGTGCCCTGGTTCGCAACCCCAAGAT CCTTCTGCTGGATGAGGCCACGTCAGCATTGGACACAGAAAGTGAAGCTGAGGTACAG GCAGCTCTGGATAAGGCCAGAGAAGGCCGGACCACCATTGTGATAGCACACCGACTGT CTACGGTCCGAAATGCAGATGTCATCGCTGGGTTTGAGGATGGAGTAATTGTGGAGCAAGGAAGCCACAGCGAACTGATGAAGAAGGAAGGGGTGTACTTCAAACTTGTCAACATGC AGACATCAGGAAGCCAGATCCAGTCAGAAGAATTTGAACTAAATGATGAAAAGGCTGCC ACTAGAATGGCCCCAAATGGCTGGAAATCTCGCCTATTTAGGCATTCTACTCAGAAAAA CCTTAAAAATTCACAAATGTGTCAGAAGAGCCTTGATGTGGAAACCGATGGACTTGAAG CAAATGTGCCACCAGTGTCCTTTCTGAAGGTCCTGAAACTGAATAAAACAGAATGGCCC TACTTTGTCGTGGGAACAGTATGTGCCATTGCCAATGGGGGGCTTCAGCCGGCATTTTC AGTCATATTCTCAGAGATCATAGCGATTTTTGGACCAGGCGATGATGCAGTGAAGCAGC AGAAGTGCAACATATTCTCTTTGATTTTCTTATTTCTGGGAATTATTTCTTTTTTTACTTTC TTCCTTCAGGGTTTCACGTTTGGGAAAGCTGGCGAGATCCTCACCAGAAGACTGCGGTC AATGGCTTTTAAAG CAATGCTAAGACAGGACATGAGCTGGTTTGATGACCATAAAAACAGTACTGGTGCACTT TCTACAAGACTTGCCACAGATGCTGCCCAAGTCCAAGGAGCCACAGGAACCAGGTTGG CTTTAATTGCACAGAATATAGCTAACCTTGGAACTGGTATTATCATATCATTTATCTACGG TTGGCAGTTAACCCTATTGCTATTAGCAGTTGTTCCAATTATTGCTGTGTCAGGAATTGT TGAAATGAAATTGTTGGCTGGAAATGCCAAAAGAGATAAAAAAGAACTGGAAGCTGCTG GAAAGATTGCAACAGAGGCAATAGAAAATATTAGGACAGTTGTGTCTTTGACCCAGGAA AGAAAATTTGAATCAATGTATGTTGAAAAATTGTATGGACCTTACAGGAATTCTGTGCAG AAGGCACACATCTATGGAATTACTTTTAGTATCTCACAAGCATTTATGTATTTTTCCTATG CCGGTTGTTTTCGATTTGGTGCATATCTCATTGTGAATGGACATATGCGCTTCAGAGATG TTATTCTGGTGTTTTCTGCAATTGTATTTGGTGCAGTGGCTCTAGGACATGCCAGTTCAT TTGCTCCAGACTATGCTAAAGCTAAGCTGTCTGCAGCCCACTTATTCATGCTGTTTGAAA GACAACCTCTGATTGACAGCTACAGTGAAGAGGGGCTGAAGCCTGATAAATTTGAAGGA AATATAACATTTAATGAAGTCGTGTTCAACTATCCCACCCGAGCAAACGTGCCAGTGCTT CAGGGGCTGAGCCTGGAGGTGAAGAAAGGCCAGACACTAGCCCTGGTGGGCAGCAGT GGCTGTGGGAAGAGCACGGTGGTCCAGCTCCTGGAGCGGTTCTACGACCCCTTGGCG GGGACAGTGCTTCTCGATGGTCAAGAAGCAAAGAAACTCAATGTCCAGTGGCTCAGAG CTCAACTCGGAATCGTGTCTCAGGAGCCTATCCTATTTGACTGCAGCATTGCCGAGAAT ATTGCCTATGGAGACAACAGCCGGGTTGTATCACAGGATGAAATTGTGAGTGCAGCCAA AGCTGCCAACATACATCCTTTCATCGAGACGTTACCCCACAAATATGAAACAAGAGTGG GAGATAAGGGGACTCAGCTCTCAGGAGGTCAAAAACAGAGGATTGCTATTGCCCGAGC CCTCATCAGACAACCTCAAATCCTCCTGTTGGATGAAGCTACATCAGCTCTGGATACTG AAAGTGAAAAGGTTGTCCAAGAAGCCCTGGACAAAGCCAGAGAAGGCCGCACCTGCAT TGTGATTGCTCACCGCCTGTCCACCATCCAGAATGCAGACTTAATAGTGGTGTTTCAGA ATGGGAGAGTCAAGGAGCATGGCACGCATCAGCAGCTGCTGGCACAGAAAGGCATCTA TTTTTCAATGGTCAGTGTCCAGGCTGGGACACAGAACTTATGA SEQ ID NO: 44 - Codon optimised human ABCB4 coding sequence (CDS) ATGGATCTGGAAGCCGCCAAGAATGGCACCGCTTGGAGGCCTACATCTGCCGAGGGC GATTTTGAGCTGGGCATCAGCAGCAAGCAGAAGCGGAAAAAGACCAAGACCGTGAAGA TGATCGGCGTGCTGACCCTGTTCCGGTACAGCGACTGGCAGGACAAGCTGTTCATGAG CCTGGGCACCATCATGGCTATCGCCCACGGATCTGGACTGCCCCTGATGATGATCGTG TTCGGCGAGATGACCGACAAGTTCGTGGACACCGCCGGCAACTTCAGCTTCCCCGTGA ATTTCAGCCTGAGCCTGCTGAACCCCGGCAAGATCCTGGAAGAGGAAATGACCAGATA CGCCTACTACTACAGCGGCCTTGGAGCTGGCGTTCTGGTGGCCGCCTATATCCAGGTG TCCTTTTGGACACTGGCCGCTGGCCGGCAGATCAGAAAGATCCGGCAGAAGTTCTTCC ACGCCATCCTGCGGCAAGAGATCGGATGGTTCGACATCAACGACACCACCGAGCTGAA CACCAGACTGACCGACGACATCAGCAAGATCAGCGAAGGCATCGGCGACAAAGTGGGA ATGTTCTTCCAGGCCGTGGCCACCTTCTTCGCCGGCTTTATCGTGGGCTTCATCAGAGG CTGGAAGCTGACCCTCGTGATCATGGCCATCTCTCCCATCCTGGGACTGTCTGCTGCTG TGTGGGCCAAGATTCTGAGCGCCTTCAGCGACAAAGAGCTGGCCGCTTACGCTAAAGC TGGCGCCGTTGCTGAAGAAGCCCTGGGAGCCATCAGAACCGTGATCGCCTTTGGCGGC CAGAACAAAGAACTGGAAAGATACCAGAAGCACCTCGAGAACGCCAAAGAGATTGGCA TCAAGAAGGCCATCAGCGCCAACATCTCTATGGGAATCGCCTTCCTGCTGATCTACGCC AGCTACGCCCTGGCCTTTTGGTACGGCTCTACCCTGGTCATCAGCAAAGAGTACACCAT CGGCAACGCCATGACCGTGTTCTTCTCCATCCTGATCGGCGCCTTTAGCGTCGGACAGGCCGCTCCTTGTATCGACGCCTTTGCTAATGCCAGAGGCGCCGCATACGTGATCTTCGA TATCATCGACAACAACCCCAAGATCGACAGCTTCAGCGAGCGGGGCCACAAGCCTGAT AGCATCAAGGGCAACCTGGAATTCAACGACGTGCACTTCAGCTACCCTAGCCGGGCCA ACGTGAAGATCCTGAAGGGCCTGAACCTGAAGGTGCAGAGCGGACAGACAGTGGCCCT CGTGGGATCTTCTGGCTGTGGCAAGAGCACCACCGTGCAGCTGATCCAGAGACTGTAC GACCCTGACGAGGGCACAATCAACATCGACGGCCAGGACATCCGGAACTTCAACGTGA ACTACCTGCGCGAGATCATCGGAGTGGTGTCCCAAGAGCCTGTGCTGTTCAGCACCAC AATCGCCGAGAACATCTGCTACGGCAGAGGCAACGTGACCATGGACGAGATCAAGAAA GCCGTGAAAGAGGCCAACGCCTACGAGTTCATCATGAAGCTGCCCCAGAAATTCGACA CCCTTGTGGGAGAAAGAGGCGCCCAACTGTCTGGCGGACAGAAGCAGAGAATCGCCAT TGCCAGAGCACTCGTGCGGAACCCTAAGATCCTGCTGCTGGACGAAGCCACAAGCGCC CTGGATACAGAGTCTGAGGCAGAAGTGCAGGCTGCCCTGGACAAGGCTAGAGAGGGC AGAACCACCATCGTGATTGCCCACAGACTGTCCACCGTGCGGAATGCCGATGTGATCG CCGGATTCGAAGATGGCGTGATCGTGGAACAGGGCTCCCACAGCGAGCTGATGAAGAA AGAAGGCGTGTACTTCAAGCTGGTCAACATGCAGACCAGCGGCTCTCAGATCCAGAGC GAGGAATTCGAGCTGAATGACGAGAAGGCCGCCACCAGGATGGCCCCTAATGGCTGGA AGTCCCGGCTGTTCAGACACAGCACCCAGAAGAATCTGAAGAACAGCCAGATGTGCCA GAAGTCCCTGGACGTCGAGACAGACGGACTGGAAGCCAATGTGCCTCCTGTGTCCTTC CTGAAAGTGCTGAAACTGAACAAGACCGAGTGGCCCTACTTCGTCGTGGGCACCGTGT GTGCCATTGCTAATGGTGGACTGCAGCCCGCCTTCTCCGTGATCTTTAGCGAGATTATC GCCATCTTTGGCCCTGGCGACGACGCCGTGAAGCAGCAGAAGTGTAATATCTTTAGCCT GATCTTCCTGTTCCTGGGGATTATCAGCTTCTTTACATTCTTCCTGCAAGGCTTCACCTT CGGCAAGGCCGGGGAGATCCTGACTCGGAGACTGAGATCCATGGCCTTCAAAGCCATG CTGAGACAGGACATGAGTTGGTTCGACGACCACAAGAACAGCACTGGCGCCCTGTCTA CCAGACTGGCCACAGATGCTGCTCAGGTGCAAGGCGCTACAGGCACAAGACTGGCCCT GATCGCCCAGAATATCGCCAATCTCGGCACCGGCATCATCATTAGCTTCATCTACGGAT GGCAGCTGACACTGCTCCTGCTGGCCGTGGTGCCTATCATTGCCGTGTCCGGCATCGT GGAAATGAAGTTGCTGGCCGGAAACGCCAAGCGGGACAAGAAAGAACTTGAGGCCGC AGGCAAGATCGCCACCGAGGCCATCGAGAATATCCGGACCGTGGTGTCTCTGACCCAA GAGCGGAAGTTCGAGTCTATGTACGTGGAAAAGCTGTACGGCCCCTACAGAAACAGCG TGCAGAAGGCCCACATCTACGGCATCACCTTCAGCATCAGCCAGGCCTTCATGTACTTT AGCTACGCCGGCTGCTTTAGATTCGGCGCCTACCTGATCGTGAACGGCCACATGAGAT TCCGGGACGTGATCCTGGTGTTCTCCGCCATCGTGTTTGGAGCCGTGGCTCTGGGACA TGCCTCTAGCTTTGCCCCTGACTACGCCAAGGCCAAGCTGTCTGCCGCACACCTGTTCA TGCTGTTCGAGAGACAGCCCCTGATCGACTCCTACAGCGAGGAAGGCCTGAAGCCAGA TAAGTTCGAGGGCAACATCACCTTTAACGAGGTGGTGTTCAACTACCCCACCAGAGCCA ACGTGCCAGTTCTGCAGGGACTGAGCCTGGAAGTGAAGAAGGGCCAGACTCTGGCTCT CGTGGGCTCTAGCGGATGTGGCAAGTCTACAGTGGTGCAGCTGCTGGAACGGTTCTAC GATCCTCTGGCTGGAACCGTCCTGCTGGATGGCCAAGAAGCCAAAAAGCTGAACGTGC AGTGGCTGAGGGCTCAGCTGGGAATCGTGTCTCAAGAACCCATCCTGTTCGACTGCAG CATTGCCGAGAATATTGCCTATGGCGACAACAGCAGAGTCGTGTCCCAGGACGAGATT GTGTCCGCTGCCAAGGCTGCCAACATTCACCCCTTCATCGAGACACTGCCCCACAAATA CGAGACAAGAGTGGGCGACAAGGGCACACAGCTGAGCGGAGGCCAGAAACAGAGGAT TGCTATCGCTCGCGCCCTGATCAGACAGCCTCAGATTCTGCTCCTGGATGAGGCCACC TCTGCTCTGGACACCGAGAGCGAGAAGGTGGTGCAAGAGGCTCTGGATAAGGCCCGC GAGGGAAGAACCTGTATCGTGATCGCTCACCGGCTGAGCACCATCCAGAATGCCGACC TCATCGTGGTGTTCCAGAACGGCAGAGTGAAAGAGCACGGCACCCACCAGCAACTGCT GGCCCAGAAGGGCATCTACTTCTCCATGGTGTCTGTGCAGGCCGGCACACAGAACCTG TGA SEQ ID NO: 45 - LPE promoter containing the LPa sequence and enhancer region deriving from human ABCB4 gene TCCCCTAGACTCTGGTTCCACCGATGCAGCCTCAGGCAGGACTTCACCTCTCTGGGCAT CCGTTTCTTCATATGTTAAACATACGGGGTTTTAATTAGATGATCGCTGAAGACCCCTCT AGCCCTAAAACTCTGTGTCTCTTAAGTGCTAAGAGGGCACCAACAGCGTTCCTCCTCCCCAAGGAGCATAATGTGATGGTTCCTGCCGGCCCTGGCTGACTCTCGCCGTCCTTGGAG ATAATTGGGTTCAGTGCCACCTGGACCAGAACTGGGGATGCGGAAGCAAGACCTCGAG GGGGTGGCCCATCAGGCGAGGCTTTCTCGGCACCCGAGGCTCCAGCCTGATCTCGGT CTTATCCTGCGACCGGGCTGGTTCTGGCGGGTCGCCAGGGTGGGCGGCGGCCCCAGC CGGGCGCCCCGGCGGCAAGAGCGGCAGGCTGCGCCCCTGGCCCGCGCCTAGCCTGG GGAGAGAGCTGGGCGGGCGGCGGGAGCTGCTCTCGCGGGCCGCGGCCCTCGCCCTG GCTGCAACGGTAGGCGTTTCCCGGGCCGGACGCGCGTGGGGGGCGGGGGCGGGGG CGGGGGCGAGGCCGCGGCGAGCAAAGTCCAGGCCCCTCTGCTGCAGCGCCCGCGCG TCCAGAGGCCCTGCCAGACACGCGCGAGGTTCGAGCCGAGAGAGGTCCGGGCGCGTC TGGCCTCGAAGGGAGACCCGGGACGTGGGGCGCGGGGCGGGAGTGGCCGGACCTCC ACCCAGTGCCCCCGGGCCCCGCGACTCGTGCGCCGGGCCGCCGGAGAGGGTGTACTT GGTTCTGAGGCTGTGGTTTCTCCTCCCGCTGAG SEQ ID NO: 46 - RRE_Rev response element of human immunodeficiency virus type 1 AGGAGCTTTGTTCCTTGGGTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCA ATGACGCTGACGGTACAGGCCAGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAA TTTGCTGAGGGCTATTGAGGCGCAACAGCATCTGTTGCAACTCACAGTCTGGGGCATCA AGCAGCTCCAGGCAAGAATCCTGGCTGTGGAAAGATACCTAAAGGATCAACAGCTCCT SEQ ID NO: 47 - WPRE sense AATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTC CTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTAT GGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTG GCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACT GGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCC TATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCG GCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAGCTGACGTCCTTTCCATGG CTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTC GGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTT CCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGC SEQ ID NO: 48 - Luciferase. GFP dual reporter gene system GACTACAAAGACCATGACGGTGATTATAAAGATCATGACATCGATTACAAGGATGACGA TGACAAGAGCGGCGGCGGCAGCATGGAGGACGCCAAGAACATCAAGAAGGGACCAGC CCCCAGATACCCCCTGGAGGACGGCACAGCCGGCGAGCAGCTGCACAAGGCCATGAA GCGGTACGCCCAGGTGCCAGGCACCATCGCCTTCACCGACGCCCACATCGAGGTGAA CATCACCTACGCCGAGTACTTCGAGATGAGCGTGCGGCTGGCCGAGGCCATGAAGCG GTACGGCCTGAACACCAACCACCGGATCGTGGTGTGCAGCGAGAACAGCCTGCAGTTC TTCATGCCCGTGCTGGGAGCCCTGTTCATCGGCGTGGCCGTGGCCCCAGCCAACGACA TCTACAACGAGCGGGAGCTGCTGAACAGCATGAACATCAGCCAGCCCACCGTGGTGTT CGTGAGCAAGAAGGGCCTGCAGAAGATCCTGAATGTGCAGAAGAAGCTGCCCATCATC CAGAAGATCATCATCATGGACAGCAAGACCGATTACCAGGGCTTCCAGAGCATGTACAC CTTCGTGACCAGCCACCTGCCCCCAGGCTTCAACGAGTACGACTTCAAGCCCGAGAGC TTCGACCGGGACAAGACCATCGCCCTGATCATGAACAGCAGCGGCAGCACCGGCCTGC CCAAGGGCGTGGCCCTGCCCCACCGGACCGCCTGCGTGCGGTTCAGCCACGCCAGAG ACCCCATCTTCGGCAACCAGATCAAGCCCGACACCGCCATCCTGAGCGTGGTGCCCTT CCACCACGGCTTCGGCATGTTCACCACCCTGGGCTACCTGATCTGCGGCTTCCGGGTG GTGCTGATGTACAGGTTCGAGGAGGAGCTGTTCCTGCGGAGCCTGCAGGACTACAAGA TCCAGAGCGCCCTGCTGGTGCCCACCCTGTTCAGCTTCTTCGCCAAGAGCACCCTGAT CGACAAGTACGACCTGAGCAACCTGCACGAGATCGCCTCTGGCGGAGCCCCACTGAGC AAGGAGGTGGGCGAGGCCGTGGCCAAGCGGTTCCACCTGCCAGGCATCCGGCAGGG CTACGGCCTGACCGAGACCACCAGCGCCATCCTGATCACCCCCGAGGGCGACGACAA GCCCGGAGCCGTGGGCAAGGTGGTGCCCTTCTTCGAGGCCAAGGTGGTGGACCTGGA CACCGGCAAGACCCTGGGCGTGAACCAGAGAGGCGAGCTGTGCGTGAGAGGCCCCAT GATCATGAGCGGCTACGTGAACAACCCCGAGGCCACCAACGCCCTGATCGACAAGGACGGCTGGCTGCACAGCGGCGACATCGCCTACTGGGACGAGGACGAGCACTTCTTCATCG TGGACCGGCTGAAGAGCCTGATCAAGTACAAGGGCTACCAGGTGGCCCCAGCCGAGC TGGAGAGCATCCTGCTGCAGCACCCCAACATCCGGGACGCCGGAGTGGCCGGACTGC CCGACGACGACGCCGGAGAGCTGCCAGCCGCCGTGGTGGTGCTGGAGCACGGCAAG ACCATGACCGAGAAGGAGATCGTGGACTACGTGGCCAGCCAGGTGACCACCGCCAAG AAGCTGAGAGGCGGCGTGGTGTTCGTGGACGAGGTGCCCAAGGGCCTGACCGGCAAG CTGGACGCCAGAAAGATCCGGGAGATCCTGATCAAGGCCAAGAAGGGCGGCAAGATC CGCGCAGAGGGCCGGGGCTCATTGCTGACCTGTGGAGATGTCGAGGAAAATCCCGGC CCCATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAG CTGGACGGCGACGTAAACGGCCACAAGTTCAGCGTGTCCGGCGAGGGCGAGGGCGAT GCCACCTACGGCAAGCTGACCCTGAAGTTCATCTGCACCACCGGCAAGCTGCCCGTGC CCTGGCCCACCCTCGTGACCACCCTGACCTACGGCGTGCAGTGCTTCAGCCGCTACCC CGACCACATGAAGCAGCACGACTTCTTCAAGTCCGCCATGCCCGAAGGCTACGTCCAG GAGCGCACCATCTTCTTCAAGGACGACGGCAACTACAAGACCCGCGCCGAGGTGAAGT TCGAGGGCGACACCCTGGTGAACCGCATCGAGCTGAAGGGCATCGACTTCAAGGAGG ACGGCAACATCCTGGGGCACAAGCTGGAGTACAACTACAACAGCCACAACGTCTATATC ATGGCCGACAAGCAGAAGAACGGCATCAAGGTGAACTTCAAGATCCGCCACAACATCG AGGACGGCAGCGTGCAGCTCGCCGACCACTACCAGCAGAACACCCCCATCGGCGACG GCCCCGTGCTGCTGCCCGACAACCACTACCTGAGCACCCAGTCCGCCCTGAGCAAAGA CCCCAACGAGAAGCGCGATCACATGGTCCTGCTGGAGTTCGTGACCGCCGCCGGGAT CACTCTCGGCATGGACGAGCTGTACAAGTAA SEQ ID NO: 49 - reverse complement of Luciferase. GFP TTACTTGTACAGCTCGTCCATGCCGAGAGTGATCCCGGCGGCGGTCACGAACTCCAGC AGGACCATGTGATCGCGCTTCTCGTTGGGGTCTTTGCTCAGGGCGGACTGGGTGCTCA GGTAGTGGTTGTCGGGCAGCAGCACGGGGCCGTCGCCGATGGGGGTGTTCTGCTGGT AGTGGTCGGCGAGCTGCACGCTGCCGTCCTCGATGTTGTGGCGGATCTTGAAGTTCAC CTTGATGCCGTTCTTCTGCTTGTCGGCCATGATATAGACGTTGTGGCTGTTGTAGTTGTA CTCCAGCTTGTGCCCCAGGATGTTGCCGTCCTCCTTGAAGTCGATGCCCTTCAGCTCGA TGCGGTTCACCAGGGTGTCGCCCTCGAACTTCACCTCGGCGCGGGTCTTGTAGTTGCC GTCGTCCTTGAAGAAGATGGTGCGCTCCTGGACGTAGCCTTCGGGCATGGCGGACTTG AAGAAGTCGTGCTGCTTCATGTGGTCGGGGTAGCGGCTGAAGCACTGCACGCCGTAGG TCAGGGTGGTCACGAGGGTGGGCCAGGGCACGGGCAGCTTGCCGGTGGTGCAGATGA ACTTCAGGGTCAGCTTGCCGTAGGTGGCATCGCCCTCGCCCTCGCCGGACACGCTGAA CTTGTGGCCGTTTACGTCGCCGTCCAGCTCGACCAGGATGGGCACCACCCCGGTGAAC AGCTCCTCGCCCTTGCTCACCATGGGGCCGGGATTTTCCTCGACATCTCCACAGGTCA GCAATGAGCCCCGGCCCTCTGCGCGGATCTTGCCGCCCTTCTTGGCCTTGATCAGGAT CTCCCGGATCTTTCTGGCGTCCAGCTTGCCGGTCAGGCCCTTGGGCACCTCGTCCACG AACACCACGCCGCCTCTCAGCTTCTTGGCGGTGGTCACCTGGCTGGCCACGTAGTCCA CGATCTCCTTCTCGGTCATGGTCTTGCCGTGCTCCAGCACCACCACGGCGGCTGGCAG CTCTCCGGCGTCGTCGTCGGGCAGTCCGGCCACTCCGGCGTCCCGGATGTTGGGGTG CTGCAGCAGGATGCTCTCCAGCTCGGCTGGGGCCACCTGGTAGCCCTTGTACTTGATC AGGCTCTTCAGCCGGTCCACGATGAAGAAGTGCTCGTCCTCGTCCCAGTAGGCGATGT CGCCGCTGTGCAGCCAGCCGTCCTTGTCGATCAGGGCGTTGGTGGCCTCGGGGTTGTT CACGTAGCCGCTCATGATCATGGGGCCTCTCACGCACAGCTCGCCTCTCTGGTTCACG CCCAGGGTCTTGCCGGTGTCCAGGTCCACCACCTTGGCCTCGAAGAAGGGCACCACCT TGCCCACGGCTCCGGGCTTGTCGTCGCCCTCGGGGGTGATCAGGATGGCGCTGGTGG TCTCGGTCAGGCCGTAGCCCTGCCGGATGCCTGGCAGGTGGAACCGCTTGGCCACGG CCTCGCCCACCTCCTTGCTCAGTGGGGCTCCGCCAGAGGCGATCTCGTGCAGGTTGCT CAGGTCGTACTTGTCGATCAGGGTGCTCTTGGCGAAGAAGCTGAACAGGGTGGGCACC AGCAGGGCGCTCTGGATCTTGTAGTCCTGCAGGCTCCGCAGGAACAGCTCCTCCTCGA ACCTGTACATCAGCACCACCCGGAAGCCGCAGATCAGGTAGCCCAGGGTGGTGAACAT GCCGAAGCCGTGGTGGAAGGGCACCACGCTCAGGATGGCGGTGTCGGGCTTGATCTG GTTGCCGAAGATGGGGTCTCTGGCGTGGCTGAACCGCACGCAGGCGGTCCGGTGGGG CAGGGCCACGCCCTTGGGCAGGCCGGTGCTGCCGCTGCTGTTCATGATCAGGGCGATGGTCTTGTCCCGGTCGAAGCTCTCGGGCTTGAAGTCGTACTCGTTGAAGCCTGGGGGC AGGTGGCTGGTCACGAAGGTGTACATGCTCTGGAAGCCCTGGTAATCGGTCTTGCTGT CCATGATGATGATCTTCTGGATGATGGGCAGCTTCTTCTGCACATTCAGGATCTTCTGCA GGCCCTTCTTGCTCACGAACACCACGGTGGGCTGGCTGATGTTCATGCTGTTCAGCAG CTCCCGCTCGTTGTAGATGTCGTTGGCTGGGGCCACGGCCACGCCGATGAACAGGGCT CCCAGCACGGGCATGAAGAACTGCAGGCTGTTCTCGCTGCACACCACGATCCGGTGGT TGGTGTTCAGGCCGTACCGCTTCATGGCCTCGGCCAGCCGCACGCTCATCTCGAAGTA CTCGGCGTAGGTGATGTTCACCTCGATGTGGGCGTCGGTGAAGGCGATGGTGCCTGG CACCTGGGCGTACCGCTTCATGGCCTTGTGCAGCTGCTCGCCGGCTGTGCCGTCCTCC AGGGGGTATCTGGGGGCTGGTCCCTTCTTGATGTTCTTGGCGTCCTCCATGCTGCCGC CGCCGCTCTTGTCATCGTCATCCTTGTAATCGATGTCATGATCTTTATAATCACCGTCAT GGTCTTTGTAGTC

[0232] Sequences of Example constructs

[0233] SEQ ID NO: 28 - CCL. LP1,coABCB4_v1 (Comparative Example)

[0234] Version 1 backbone containing LP1 promoter CCATTGCATACGTTGTATCCATATCATAATATGTACATTTATATTGGCTCATGTCCAACAT TACCGCCATGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATT AGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTG GCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTA ACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCA CTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACG GTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGG CAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATC AATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGT CAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTC CGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGA GCTCGTTTAGTGAACCGGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCT GGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGT AGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGT CAGTGTGGAAAATCTCTAGCAGTGGCGCCCGAACAGGGACTTGAAAGCGAAAGGGAAA CCAGAGGAGCTCTCTCGACGCAGGACTCGGCTTGCTGAAGCGCGCACGGCAAGAGGC GAGGGGCGGCGACTGGTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGAAGGAGA GAGATGGGTGCGAGAGCGTCAGTATTAAGCGGGGGAGAATTAGATCGCGATGGGAAAA AATTCGGTTAAGGCCAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTATGGGCAA GCAGGGAGCTAGAACGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGT AGACAAATACTGGGACAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATC ATTATATAATACAGTAGCAACCCTCTATTGTGTGCATCAAAGGATAGAGATAAAAGACAC CAAGGAAGCTTTAGACAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCACAG CAAGCGGCCGCTGATCTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAGT GAATTATATAAATATAAAGTAGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGCA AAGAGAAGAGTGGTGCAGAGAGAAAAAAGAGCAGTGGGAATAGGAGCTTTGTTCCTTG GGTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACA GGCCAGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTG AGGCGCAACAGCATCTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAG AATCCTGGCTGTGGAAAGATACCTAAAGGATCAACAGCTCCTGGGGATTTGGGGTTGCT CTGGAAAACTCATTTGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTC TGGAACAGATTTGGAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTAC ACAAGCTTAATACACTCCTTAATTGAAGAATCGCAAAACCAGCAAGAAAAGAATGAACAA GAATTATTGGAATTAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACATAACAAATTGG CTGTGGTATATAAAATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTT TTGCTGTACTTTCTATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGA CCCACCTCCCAACCCCGAGGGGACCCGACAGGCCCGAAGGAATAGAAGAAGAAGGTG GAGAGAGAGACAGAGACAGATCCATTCGATTAGTGAACGGATCTCGACGGTATCGGTTAACTTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGACA TAATAGCAACAGACATACAAACTAAAGAATTACAAAAACAAATTACAAAAATTCAAAATTT TATCGATCACGAGACTAGCCTCGAGGATATCCCCTAAAATGGGCAAACATTGCAAGCA GCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGT CAGAGACCTCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCG GTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGGAATGAC TCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGC GTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGCCCCTGTTTGCTCCTCCGAT AACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATC CACTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCA CTGACCTGGGACAGTGAATCACCGGTGATCCGCCACCATGGATCTGGAAGCCGCCAA GAA TGGCA CCGCTTGGA GGCCTA CA TCTGCCGA GGGCGA TTTTGAGCTGGGCA TCA G CAGCAA GCA GAAGCGGAAAAA GA CCA A GA CCGTGAA GA TGA TCGGCGTGCTGA CCC TGTTCCGGTA CA GCGA CTGGCAGGA CAA GCTGTTCA TGA GCCTGGGCACCA TCA TGG CT A TCGCCCACGGA TCTGGA CTGCCCCTGA TGA TGA TCGTGTTCGGCGA GA TGA CCG A CAAGTTCGTGGA CA CCGCCGGCAA CTTCA GCTTCCCCGTGAA TTTCAGCCTGA GCCT GCTGAACCCCGGCAAGATCCTGGAAGAGGAAATGACCAGATACGCCTACTACTACAG CGGCCTTGGAGCTGGCGTTCTGGTGGCCGCCTA TA TCCA GGTGTCCTTTTGGA CA CTG GCCGCTGGCCGGCAGATCAGAAAGATCCGGCAGAAGTTCTTCCACGCCATCCTGCGG CAA GA GA TCGGA TGGTTCGA CA TCA A CGA CA CCA CCG A GCTGAACA CCA GA CTG A CC GA CGA CA TCAGCAA GA TCA GCGA A GGCA TCGGCGA CAAAGTGGGAA TGTTCTTCCA G GCCGTGGCCACCTTCTTCGCCGGCTTTATCGTGGGCTTCATCAGAGGCTGGAAGCTGA CCCTCGTGA TCA TGGCCA TCTCTCCCA TCCTGGGA CTGTCTGCTGCTGTGTGGGCCAA GA TTCTGA GCGCCTTCAGCGA CAAA GA GCTGGCCGCTTACGCTAAAGCTGGCGCCGT TGCTGA A GA AGCCCTGGGA GCCA TCA GAA CCGTGA TCGCCTTTGGCGGCCA GAA CAA A GAACTGGAAA GA TACCA GAA GCA CCTCGA GAA CGCCAAA GA GA TTGGCA TCA AG A A GGCCA TCAGCGCCAA CA TCTCTA TGGGAA TCGCCTTCCTGCTGA TCTA CGCCA GCTA CGCCCTGGCCTTTTGGTACGGCTCTACCCTGGTCATCAGCAAAGAGTACACCATCGGC A A CGCCA TGA CCGTGTTCTTCTCCA TCCTGA TCGGCGCCTTTA GCGTCGGA CA GGCCG CTCCTTGTA TCGA CGCCTTTGCTAA TGCCA GA GGCGCCGCA TA CGTGA TCTTCGA TA T CA TCGA C A AC A A CCCCAA GA TCGA CA GCTTCAGCGA GCGGGGCCA CAA GCCTGA TA GCATCAAGGGCAACCTGGAATTCAACGACGTGCACTTCAGCTACCCTAGCCGGGCCA A CGTGA AG A TCCTGA A GGGCCTGAA CCTGAA GGTGCAGA GCGGA CA GA CA GTGGCC CTCGTGGGA TCTTCTGGCTGTGGCAA GA GCA CCACCGTGCA GCTGA TCCA GA GA CTG TA CGA CCCTGA CGA GGGCA CAA TCA A CA TCGA CGGCCA GGA CA TCCGGAA CTTCA A C GTGAACTACCTGCGCGA GA TCA TCGGA GTGGTGTCCCAA GA GCCTGTGCTGTTCA GC A CCA CAA TCGCCGA GA AC A TCTGCTACGGCA GA GGCA A CGTGA CCA TGG A CGA GA TC AAGAAA GCCGTGAAAGA GGCCA ACGCCTACGA GTTCA TCA TGAAGCTGCCCCA GAAA TTCGACACCCTTGTGGGAGAAA GA GGCGCCCAA CTGTCTGGCGGACAGA AGCAGA G A A TCGCCA TTGCCA GA GCACTCGTGCGGAA CCCTAA GA TCCTGCTGCTGGA CGA A GC CA CAA GCGCCCTGGA TA CA GAGTCTGAGGCA GAAGTGCAGGCTGCCCTGGA CAAGG CT A GA GA GGGCAGAACCA CCA TCGTGA TTGCCCA CA GA CTGTCCA CCGTGCGGAA TG CCG A TGTGA TCGCCGGA TTCGAAGA TGGCGTGA TCGTGGAACAGGGCTCCCACAGCG A GCTGA TGA AG AAA GAAGGCGTGTA CTTCA A GCTGGTCAACA TGC A GA CCAGCGGCT CTCA GA TCCA GA GCGA GGA A TTCGAGCTGAA TGA CGA GAA GGCCGCCA CCA GGA TG GCCCCTAA TGGCTGGAAGTCCCGGCTGTTCA GA CA CA GCA CCC A GAA GAA TCTGAA G AA CA GCCA GA TGTGCCA GAAGTCCCTGGA CGTCGA GA CA GA CGGA CTGGAA GCCAA TGTGCCTCCTGTGTCCTTCCTGAAAGTGCTGAAACTGAACAAGACCGAGTGGCCCTAC TTCGTCGTGGGCACCGTGTGTGCCATTGCTAATGGTGGACTGCAGCCCGCCTTCTCCG TGATCTTTAGCGAGATTATCGCCATCTTTGGCCCTGGCGACGACGCCGTGAAGCAGCA GAA GTGTAA TA TCTTTA GCCTGA TCTTCCTGTTCCTGGGGA TTA TCAGCTTCTTTACA TT CTTCCTGCAA GGCTTCA CCTTCGGCAA GGCCGGGGA GA TCCTGA CTCGGAGA CTG A G A TCCA TGGCCTTCAAA GCCA TGCTGA GA CA GGA CA TGAGTTGGTTCGA CGA CCACAA GAA CA GCACTGGCGCCCTGTCTACCA GA CTGGCCA CA GA TGCTGCTCA GGTGCAA GG CGCTA CA GGCACAA GA CTGGCCCTGA TCGCCCA GAA TA TCGCCA A TCTCGGCA CCGGCA TCA TCA TTAGCTTCA TCTA CGGA TGGCAGCTGA CA CTGCTCCTGCTGGCCGTGGTG CCT A TCA TTGCCGTGTCCGGCA TCGTGGAAA TGAAGTTGCTGGCCGGAAA CGCCAAG CGGGA CAAGAAAGAA CTTGA GGCCGCA GGCAA GA TCGCCA CCGA GGCCA TCGA GAA TA TCCGGA CCGTGGTGTCTCTGA CCCAA GA GCGGAAGTTCGA GTCTA TGTA CGTGGA AAA GCTGTA CGGCCCCTA CA GAAA CA GCGTGCA GAA GGCCCA CA TCTA CGGCA TCA C CTTCA GCA TCA GCCA GGCCTTCA TGTA CTTTA GCTACGCCGGCTGCTTTA GA TTCGGC GCCTA CCTGA TCGTGAA CGGCCA CA TGA GA TTCCGGGA CGTGA TCCTGGTGTTCTCC GCCA TCGTGTTTGGA GCCGTGGCTCTGGGACA TGCCTCTA GCTTTGCCCCTGA CT A CG CCA A GGCCAAGCTGTCTGCCGCA CA CCTGTTCA TGCTGTTCGAGA GA CA GCCCCTGA TCGA CTCCTA CA GCGA GGAAGGCCTGAA GCCA GA TAAGTTCGA GGGCAA CA TCA CCT TTAA CGA GGTGGTGTTCAA CT A CCCCA CCA GA GCCAACGTGCCA GTTCTGCA GGGA C TGA GCCTGGAA GTGAA GAA GGGCCA GA CTCTGGCTCTCGTGGGCTCTA GCGGA TGTG GCAAGTCTACAGTGGTGCAGCTGCTGGAACGGTTCTACGATCCTCTGGCTGGAACCG TCCTGCTGGATGGCCAAGAAGCCAAAAAGCTGAACGTGCAGTGGCTGAGGGCTCAGC TGGGAA TCGTGTCTCAAGAACCCA TCCTGTTCGA CTGCA GCA TTGCCGA GA AT A TTGC CT A TGGCGA CAACAGCA GAGTCGTGTCCCA GGA CGA GA TTGTGTCCGCTGCCAA GGC TGCCAA CA TTCA CCCCTTCA TCGA GA CA CTGCCCCACAAATA CGA GA CAA GAGTGGG CGA CAA GGGCA CA CA GCTGAGCGGA GGCCA GAAA CA GA GGA TTGCTA TCGCTCGCG CCCTGA TCAGA CA GCCTCA GA TTCTGCTCCTGGA TGA GGCCA CCTCTGCTCTGGA CA C CGA GA GCGA GAAGGTGGTGCAA GA GGCTCTGGATAA GGCCCGCGA GGGA A GAA CCT GT A TCGTGA TCGCTCA CCGGCTGA GCA CCA TCCA GAA TGCCGA CCTCA TCGTGGTGTT CCA GAACGGCA GAGTGAAA GA GCACGGCA CCCA CCA GCA A CTGCTGGCCCA GAAGG GCA TCTACTTCTCCA TGGTGTCTGTGCAGGCCGGCA CA CA GAA CC7G7GAGTCGACAA TCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCT TTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATG GCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGG CCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTG GTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCT ATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGG CTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAGCTGACGTCCTTTCCATGGCT GCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCG GCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTC CGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCC TGGAATTCGAGCTCGGTACCTTTAAGACCAATGACTTACAAGGCAGCTGTAGATCTTAG CCACTTTTTAAAAGAAAAGGGGGGACTGGAAGGGCTAATTCACTCCCAACGAAGACAAG ATCTGCTTTTTGCTTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTC TCTGGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCA AGTAGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTT AGTCAGTGTGGAAAATCTCTAGCAGTAGTAGTTCATGTCATCTTATTATTCAGTATTTATA ACTTGCAAAGAAATGAATATCAGAGAGTGAGAGGAACTTGTTTATTGCAGCTTATAATGG TTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCT AGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGCTCTAGCTATCCCGC CCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCA TGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTAT TCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGGACGTACCCAATTCGCCCTATA GTGAGTCGTATTACGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAAC CCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAA TAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAA TGGGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGC GTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTT TCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGG TTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCA CGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTT CTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTC TTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTTAGGTGGCACTTTTCG GGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCC GCTCATGAGACAATAACCCTGATAAATGCTTCAATAATAGCACCTAGATCAAGAGACAG GATGAGGATCGTTTCGCATGATTGAACAAGATGGATTGCACGCAGGTTCTCCGGCCGCT TGGGTGGAGAGGCTATTCGGCTATGACTGGGCACAACAGACAATCGGCTGCTCTGATG CCGCCGTGTTCCGGCTGTCAGCGCAGGGGCGCCCGGTTCTTTTTGTCAAGACCGACCT GTCCGGTGCCCTGAATGAACTGCAAGACGAGGCAGCGCGGCTATCGTGGCTGGCCAC GACGGGCGTTCCTTGCGCAGCTGTGCTCGACGTTGTCACTGAAGCGGGAAGGGACTG GCTGCTATTGGGCGAAGTGCCGGGGCAGGATCTCCTGTCATCTCACCTTGCTCCTGCC GAGAAAGTATCCATCATGGCTGATGCAATGCGGCGGCTGCATACGCTTGATCCGGCTA CCTGCCCATTCGACCACCAAGCGAAACATCGCATCGAGCGAGCACGTACTCGGATGGA AGCCGGTCTTGTCGATCAGGATGATCTGGACGAAGAGCATCAGGGGCTCGCGCCAGCC GAACTGTTCGCCAGGCTCAAGGCGAGCATGCCCGACGGCGAGGATCTCGTCGTGACC CATGGCGATGCCTGCTTGCCGAATATCATGGTGGAAAATGGCCGCTTTTCTGGATTCAT CGACTGTGGCCGGCTGGGTGTGGCGGACCGCTATCAGGACATAGCGTTGGCTACCCG TGATATTGCTGAAGAGCTTGGCGGCGAATGGGCTGACCGCTTCCTCGTGCTTTACGGTA TCGCCGCTCCCGATTCGCAGCGCATCGCCTTCTATCGCCTTCTTGACGAGTTCTTCTGA ATTATTAACGCTTACAATTTCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTT CACACCGCATCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATT TTTCTAAATACATTCAAATATGTATCCGCTCATGACCAAAATCCCTTAACGTGAGTTTTCG TTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTT CTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTT GCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGA TACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAG CACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGAT AAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGT CGGGCTGAACGGGGGGGTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCG AACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAA GGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGC TTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTT GAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCA ACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTG CGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCT CGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCG CCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCAC GACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGC TCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAA TTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCGC GCAATTAACCCTCACTAAAGGGAACAAAAGCTGGAGCTGCAAGCTTGG

[0235] Annotations

[0236] • 1..73: Part of the plasmid needed for bacterial work

[0237] • 74..657: CMV promoter - allows expression of the vector genome

[0238] • 658..7806: vector genome

[0239] • 672..852: 5' LTR (truncated)- truncated 5' long terminal repeat (LTR) from HIV-1 • 899..1024: HIV-1 Ψ_ packaging signal of human immunodeficiency virus type 1

[0240] • 1517..1750: RRE_ The Rev response element (RRE) of HIV-1 allows for Rev- dependent mRNA export from the nucleus to the cytoplasm.

[0241] • 2246..2362: cPPT / CTS_ central polypurine tract and central termination sequence of HIV-1

[0242] • 2394..2841: LP1 promoter_ hApoE-HCR (Apolipoprotein E Hepatic Control Region) fused to hAAT (alpha-1 anti-trypsin) promoter (shown in bold)

[0243] • 2859..6698: coABCB4_ codon optimised version of the human ABCB4 gene (shown in bold and italics)

[0244] • 6705..7293: WPRE_ woodchuck hepatitis virus posttranscriptional regulatory element• 7380..7613: 3' LTR (AU3)_ self-inactivating 3' long terminal repeat (LTR) from HIV-1 • 7685..7806: SV40 poly(A) signal_ SV40 polyadenylation signal

[0245] • 7807..0: Part of the plasmid needed for bacterial work

[0246] SEQ ID NO: 29 - CCL. LPa. coABCB4, Version 2 backbone -coABCB4 and LPa promoter AACTTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGAC ATAATAGCAACAGACATACAAACTAAAGAATTACAAAAACAAATTACAAAAATTCAAAATT TTATCGATCACGAGACTAGCCTCGAGGGGGTGGCCCATCAGGCGAGGCTTTCTCGGCA CCCGAGGCTCCAGCCTGATCTCGGTCTTATCCTGCGACCGGGCTGGTTCTGGCGGGT CGCCAGGGTGGGCGGCGGCCCCAGCCGGGCGCCCCGGCGGCAAGAGCGGCAGGCT GCGCCCCTGGCCCGCGCCTAGCCTGGGGAGAGAGCTGGGCGGGCGGCGGGAGCTG CTCTCGCGGGCCGCGGCCCTCGCCCTGGCTGCAACGGTAGGCGTTTCCCGGGCCGG ACGCGCGTGGGGGGCGGGGGCGGGGGCGGGGGCGAGGCCGCGGCGAGCAAAGTC CAGGCCCCTCTGCTGCAGCGCCCGCGCGTCCAGAGGCCCTGCCAGACACGCGCGAG GTTCGAGCCGAGAGAGGTCCGGGCGCGTCTGGCCTCGAAGGGAGACCCGGGACGTG GGGCGCGGGGCGGGAGTGGCCGGACCTCCACCCAGTGCCCCCGGGCCCCGCGACT CGTGCGCCGGGCCGCCGGAGAGGGTGTACTTGGTTCTGAGGCTGTGGTTTCTCCTCC CGCTGAGACCGGTGATCCGCCACCATGGATCTGGAAGCCGCCAAGAATGGCACCGCT TGGA GGCCTA CA TCTGCCGA GGGCGA TTTTGA GCTGGGCA TCA GCA GCAAGCAGAAG CGGAAAAA GA CCAA GA CCGTGAA GA TGA TCGGCGTGCTGA CCCTGTTCCGGTA CA GC GA CTGGCA GGA CAAGCTGTTCA TGA GCCTGGGCA CCA TCA TGGCTA TCGCCCACGGA TCTGGA CTGCCCCTGA TGA TGA TCGTGTTCGGCGA GA TGA CCGA CAAGTTCGTGGA C ACCGCCGGCAACTTCAGCTTCCCCGTGAATTTCAGCCTGAGCCTGCTGAACCCCGGC AAGA TCCTGGAAGAGGAAA TGA CCAGA TA CGCCTACTA CTACAGCGGCCTTGGAGCT GGCGTTCTGGTGGCCGCCTA TA TCCAGGTGTCCTTTTGGA CA CTGGCCGCTGGCCGG CA GA TCA GAAA GA TCCGGCAGAAGTTCTTCCA CGCCA TCCTGCGGCAA GA GA TCGGA TGGTTCGA CA TCA A CGA CA CCACCGA GCTGAA CA CCA GA CTGA CCGA CGA CA TCAGC A A GA TCA GCGAAGGCA TCGGCGA CAAA GTGGGAATGTTCTTCCA GGCCGTGGCCA CC TTCTTCGCCGGCTTTA TCGTGGGCTTCA TCA GA GGCTGGAA GCTGA CCCTCGTGA TCA TGGCCA TCTCTCCCA TCCTGGGA CTGTCTGCTGCTGTGTGGGCCAA GA TTCTGA GCGC CTTCA GCGA CAAA GA GCTGGCCGCTTA CGCTAAAGCTGGCGCCGTTGCTGAA GAAGC CCTGGGAGCCA TCAGAA CCGTGA TCGCCTTTGGCGGCCA GA ACAA AGA ACTGGA AA G A TA CCA GAAGCACCTCGA GAA CGCCA A A GA GA TTGGCA TCA A GAA GGCCA TCA GCG CCAA CA TCTCTA TGGGA A TCGCCTTCCTGCTGA TCTA CGCCA GCTACGCCCTGGCCTT TTGGTA CGGCTCTA CCCTGGTCA TCAGCAAA GA GT A CA CCA TCGGCAA CGCCA TGA C CGTGTTCTTCTCCA TCCTGA TCGGCGCCTTTA GCGTCGGA CA GGCCGCTCCTTGTA TC GA CGCCTTTGCTAA TGCCA GA GGCGCCGCA TA CGTGA TCTTCGA TA TCA TCGA CAA CA A CCCCAA GA TCGA CA GCTTCA GCGAGCGGGGCCA CAA GCCTGA TA GCA TCA A GGGC AACCTGGAATTCAACGACGTGCACTTCAGCTACCCTAGCCGGGCCAACGTGAAGATC CTGA A GGGCCTGAA CCTGAAGGTGCA GA GCGGA CA GA CAGTGGCCCTCGTGGGA TC TTCTGGCTGTGGCAA GA GCACCA CCGTGCAGCTGA TCCAGAGA CTGTA CGA CCCTGA CGA GGGC A CAA TCA A CA TCGA CGGCCA GGA CA TCCGGAA CTTCA A CGTGA ACTA CCT GCGCGA GA TCA TCGGA GTGGTGTCCCA A GA GCCTGTGCTGTTCA GCA CCA CAA TCGC CGA GA AC A TCTGCTACGGCA GA GGCAA CGTGA CCA TGGA CGA GA TCA A GAAA GCCG TGAAA GA GGCCA A CGCCTACGA GTTCA TCA TGA A GCTGCCCCA GAAA TTCGA CA CCC TTGTGGGA GAAA GAGGCGCCCAA CTGTCTGGCGGA CA GA AGCAGAGA ATCGCCA TT GCCAGAGCACTCGTGCGGAACCCTAAGATCCTGCTGCTGGACGAAGCCACAAGCGC CCTGGA TA CA GA GTCTGAGGCAGAAGTGCAGGCTGCCCTGGA CAAGGCTAGAGAGG GCAGAACCA CCATCGTGA TTGCCCACAGA CTGTCCA CCGTGCGGAA TGCCGA TGTGA TCGCCGGA TTCGA A GA TGGCGTGA TCGTGGAA CA GGGCTCCCA CA GCGA GCTGA TGA A GAAA GAAGGCGTGTA CTTCA A GCTGGTCAA CA TGC A GA CCA GCGGCTCTCAGA TCC A GA GCGA GGAATTCGA GCTGAA TGA CGA GAA GGCCGCCA CCA GGA TGGCCCCTAA T GGCTGGAAGTCCCGGCTGTTCA GA CA CA GCACCCAGA AGA ATCTGAA GAACAGCCA GA TGTGCCA GAAGTCCCTGGA CGTCGA GA CA GA CGGA CTGGAAGCCAA TGTGCCTCCTGTGTCCTTCCTGAAAGTGCTGAAA CTGAA CAAGA CCGA GTGGCCCTACTTCGTCGTG GGCACCGTGTGTGCCA TTGCTAA TGGTGGA CTGCA GCCCGCCTTCTCCGTGA TCTTTA GCGA GA TTA TCGCCA TCTTTGGCCCTGGCGA CGA CGCCGTGAAGCA GCA GAAGTGTA A TA TCTTTA GCCTGA TCTTCCTGTTCCTGGGGA TTA TCA GCTTCTTTACA TTCTTCCTGC A A GGCTTCA CCTTCGGCAA GGCCGGGGA GA TCCTGA CTCGGA GA CTGA GA TCCA TGG CCTTCAAA GCCA TGCTGA GA CA GGA CA TGAGTTGGTTCGA CGA CCA CAA GAA CA GCA CTGGCGCCCTGTCTA CCA GA CTGGCCA CA GA TGCTGCTCAGGTGCAAGGCGCTACAG GCACAA GA CTGGCCCTGA TCGCCCA GAA TA TCGCCA A TCTCGGCA CCGGCA TCA TCA TTA GCTTCA TCTA CGGA TGGCA GCTGA CA CTGCTCCTGCTGGCCGTGGTGCCTA TCA T TGCCGTGTCCGGCATCGTGGAAATGAAGTTGCTGGCCGGAAACGCCAAGCGGGACA AGAAAGAACTTGA GGCCGCAGGCAA GA TCGCCACCGA GGCCA TCGAGAA TA TCCGG ACCGTGGTGTCTCTGACCCAAGAGCGGAAGTTCGAGTCTATGTACGTGGAAAAGCTG TACGGCCCCTACAGAAA CA GCGTGCAGAAGGCCCACATCTACGGCATCA CCTTCA GC ATCAGCCAGGCCTTCATGTACTTTAGCTACGCCGGCTGCTTTAGATTCGGCGCCTACC TGA TCGTGAA CGGCCA CA TGA GA TTCCGGGA CGTGA TCCTGGTGTTCTCCGCCA TCGT GTTTGGAGCCGTGGCTCTGGGACATGCCTCTAGCTTTGCCCCTGACTACGCCAAGGC CAA GCTGTCTGCCGCA CA CCTGTTCA TGCTGTTCGAGA GA CA GCCCCTGA TCGA CTCC TA CA GCGA GGA A GGCCTGAA GCCA GA TAA GTTCGA GGGCAA CA TCA CCTTTAA CGA G GTGGTGTTCAACTA CCCCACCA GA GCCAACGTGCCAGTTCTGCAGGGACTGA GCCTG GAA GTGAA GAA GGGCCA GA CTCTGGCTCTCGTGGGCTCTA GCGGA TGTGGCAA GTCT A CA GTGGTGCA GCTGCTGGAA CGGTTCTACGA TCCTCTGGCTGGAA CCGTCCTGCTG GA TGGCCAA GAA GCCAAAAA GCTGAACGTGCA GTGGCTGA GGGCTCA GCTGGGAA T CGTGTCTCAAGAACCCA TCCTGTTCGA CTGCA GCA TTGCCGA GAA TA TTGCCTA TGGC GA CAA CA GCAGA GTCGTGTCCCA GGA CGA GA TTGTGTCCGCTGCCAAGGCTGCCAAC A TTCA CCCCTTCA TCGA GA CA CTGCCCCA CAA A TA CGA GA CAA GA GTGGGCGA CAA G GGCACACAGCTGAGCGGAGGCCA GAAA CA GA GGA TTGCTA TCGCTCGCGCCCTGA T CA GA CA GCCTCA GA TTCTGCTCCTGGA TGAGGCCA CCTCTGCTCTGGA CA CCGA GA G CGA GAAGGTGGTGCAA GA GGCTCTGGA TAAGGCCCGCGA GGGAA GAA CCTGTA TCG TGA TCGCTCACCGGCTGA GCA CCA TCCA GAA TGCCGA CCTCA TCGTGGTGTTCCAGA A CGGCA GAGTGAAAGA GCA CGGCA CCCA CCA GCA A CTGCTGGCCCA GAA GGGCA TC TACTTCTCCATGGTGTCTGTGCAGGCCGGCACACAGAACCTGTGAGTCGACAATCAAC CTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTAC GCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTT CATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGT TGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGG GGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGC CACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTT GGGCACTGACAATTCCGTGGTGTTGTCGGGGAAGCTGACGTCCTTTCCATGGCTGCTC GCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCC TCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCG TCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCCTGGA ATTCGAGCTCGGTACCTTTAAGACCAATGACTTACAAGGCAGCTGTAGATCTTAGCCAC TTTTTAAAAGAAAAGGGGGGACTGGAAGGGCTAATTCACTCCCAACGAAGACAAGATCT GCTTTTTGCTTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTG GCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTA GTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTC AGTGTGGAAAATCTCTAGCAGTAGTAGTTCATGTCATCTTATTATTCAGTATTTATAACTT GCAAAGAAATGAATATCAGAGAGTGAGAGGAACTTGTTTATTGCAGCTTATAATGGTTAC AAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTT GTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGCTCTAGCTATCCCGCCCCT AACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATGGC TGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTATTCCA GAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGGACGTACCCAATTCGCCCTATAGTGA GTCGTATTACGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTG GCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGG GACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTG ACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCT CGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTC CGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACG TAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCT TTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTT TTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACA AAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTTAGGTGGCACTTTTCGGG GAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCT CATGAGACAATAACCCTGATAAATGCTTCAATAATAGCACCTAGATCAAGAGACAGGATG AGGATCGTTTCGCATGATTGAACAAGATGGATTGCACGCAGGTTCTCCGGCCGCTTGG GTGGAGAGGCTATTCGGCTATGACTGGGCACAACAGACAATCGGCTGCTCTGATGCCG CCGTGTTCCGGCTGTCAGCGCAGGGGCGCCCGGTTCTTTTTGTCAAGACCGACCTGTC CGGTGCCCTGAATGAACTGCAAGACGAGGCAGCGCGGCTATCGTGGCTGGCCACGAC GGGCGTTCCTTGCGCAGCTGTGCTCGACGTTGTCACTGAAGCGGGAAGGGACTGGCT GCTATTGGGCGAAGTGCCGGGGCAGGATCTCCTGTCATCTCACCTTGCTCCTGCCGAG AAAGTATCCATCATGGCTGATGCAATGCGGCGGCTGCATACGCTTGATCCGGCTACCTG CCCATTCGACCACCAAGCGAAACATCGCATCGAGCGAGCACGTACTCGGATGGAAGCC GGTCTTGTCGATCAGGATGATCTGGACGAAGAGCATCAGGGGCTCGCGCCAGCCGAAC TGTTCGCCAGGCTCAAGGCGAGCATGCCCGACGGCGAGGATCTCGTCGTGACCCATG GCGATGCCTGCTTGCCGAATATCATGGTGGAAAATGGCCGCTTTTCTGGATTCATCGAC TGTGGCCGGCTGGGTGTGGCGGACCGCTATCAGGACATAGCGTTGGCTACCCGTGATA TTGCTGAAGAGCTTGGCGGCGAATGGGCTGACCGCTTCCTCGTGCTTTACGGTATCGC CGCTCCCGATTCGCAGCGCATCGCCTTCTATCGCCTTCTTGACGAGTTCTTCTGAATTAT TAACGCTTACAATTTCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACA CCGCATCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTC TAAATACATTCAAATATGTATCCGCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCC ACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGC GCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCG GATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACC AAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACC GCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGT CGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGG CTGAACGGGGGGGTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCG GACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCA GGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCG TCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCG GCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTA TCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCG CAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAAT ACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGG TTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCA TTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGA GCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCGCGCAATT AACCCTCACTAAAGGGAACAAAAGCTGGAGCTGCAAGCTTGGCCATTGCATACGTTGTA TCCATATCATAATATGTACATTTATATTGGCTCATGTCCAACATTACCGCCATGTTGACAT TGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATA TGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGA CCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTT TCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAA GTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTG GCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATT AGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTT TGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCA AATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCGTTTAGTGAAC CGGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGGCTAACTAGGGAA CCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCCCGTC TGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAAAATCT CTAGCAGTGGCGCCCGAACAGGGACTTGAAAGCGAAAGGGAAACCAGAGGAGCTCTCT CGACGCAGGACTCGGCTTGCTGAAGCGCGCACGGCAAGAGGCGAGGGGCGGCGACT GGTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGAAGGAGAGAGATGGGTGCGAG AGCGTCAGTATTAAGCGGGGGAGAATTAGATCGCGATGGGAAAAAATTCGGTTAAGGC CAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTATGGGCAAGCAGGGAGCTAGAA CGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGTAGACAAATACTGGG ACAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATCATTATATAATACAGT AGCAACCCTCTATTGTGTGCATCAAAGGATAGAGATAAAAGACACCAAGGAAGCTTTAG ACAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCACAGCAAGCGGCCGCTGA TCTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAGTGAATTATATAAATAT AAAGTAGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGCAAAGAGAAGAGTGGT GCAGAGAGAAAAAAGAGCAGTGGGAATAGGAGCTTTGTTCCTTGGGTTCTTGGGAGCA GCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACAGGCCAGACAATTAT TGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTGAGGCGCAACAGCAT CTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAGAATCCTGGCTGTGG AAAGATACCTAAAGGATCAACAGCTCCTGGGGATTTGGGGTTGCTCTGGAAAACTCATT TGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTCTGGAACAGATTTG GAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTACACAAGCTTAATAC ACTCCTTAATTGAAGAATCGCAAAACCAGCAAGAAAAGAATGAACAAGAATTATTGGAAT TAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACATAACAAATTGGCTGTGGTATATAA AATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTTTTGCTGTACTTTC TATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGACCCACCTCCCAAC CCCGAGGGGACCCGACAGGCCCGAAGGAATAGAAGAAGAAGGTGGAGAGAGAGACAG AGACAGATCCATTCGATTAGTGAACGGATCTCGACGGTATCGGTT

[0247] Annotations

[0248] • 4..121: cPPT / CTS_ central polypurine tract and central termination sequence of HIV-1 • 147..685: LPa_ promoter (shown bold)

[0249] • 703..4542: coABCB4_ codon optimised version of the human ABCB4 gene (shown bold and in italics)

[0250] • 4549..5137: WPRE_ woodchuck hepatitis virus posttranscriptional regulatory element • 5224..5457: 3' LTR (AU3)_ self-inactivating 3' long terminal repeat (LTR) from HIV-1 • 5529..5650: SV40 poly(A) signal_ SV40 polyadenylation signal

[0251] • 5651..8788: Part of the plasmid needed for bacterial work

[0252] • 8789..9372: CMV promoter - allows expression of the vector genome

[0253] • 9387..5650: Vector genome

[0254] • 9387..9567: 5' LTR (truncated)- truncated 5' long terminal repeat (LTR) from HIV-1 • 9614..9739: HIV-1 packaging signal of human immunodeficiency virus type 1 • 10232..10465: RRE_ The Rev response element (RRE) of HIV-1 allows for Rev- dependent mRNA export from the nucleus to the cytoplasm

[0255] SEQ ID NO: 30 - CCL. LPE. COABCB4, Version 2 backbone - coABCB4 and LPE AACTTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGAC ATAATAGCAACAGACATACAAACTAAAGAATTACAAAAACAAATTACAAAAATTCAAAATT TTATCGATCACGAGACTAGTCCCCTAGACTCTGGTTCCACCGATGCAGCCTCAGGCAG GACTTCACCTCTCTGGGCATCCGTTTCTTCATATGTTAAACATACGGGGTTTTAATTAG ATGATCGCTGAAGACCCCTCTAGCCCTAAAACTCTGTGTCTCTTAAGTGCTAAGAGGG CACCAACAGCGTTCCTCCTCCCCAAGGAGCATAATGTGATGGTTCCTGCCGGCCCTG GCTGACTCTCGCCGTCCTTGGAGATAATTGGGTTCAGTGCCACCTGGACCAGAACTGGGGATGCGGAAGCAAGACCTCGAGGGGGTGGCCCATCAGGCGAGGCTTTCTCGGCA CCCGAGGCTCCAGCCTGATCTCGGTCTTATCCTGCGACCGGGCTGGTTCTGGCGGGT CGCCAGGGTGGGCGGCGGCCCCAGCCGGGCGCCCCGGCGGCAAGAGCGGCAGGCT GCGCCCCTGGCCCGCGCCTAGCCTGGGGAGAGAGCTGGGCGGGCGGCGGGAGCTG CTCTCGCGGGCCGCGGCCCTCGCCCTGGCTGCAACGGTAGGCGTTTCCCGGGCCGG ACGCGCGTGGGGGGCGGGGGCGGGGGCGGGGGCGAGGCCGCGGCGAGCAAAGTC CAGGCCCCTCTGCTGCAGCGCCCGCGCGTCCAGAGGCCCTGCCAGACACGCGCGAG GTTCGAGCCGAGAGAGGTCCGGGCGCGTCTGGCCTCGAAGGGAGACCCGGGACGTG GGGCGCGGGGCGGGAGTGGCCGGACCTCCACCCAGTGCCCCCGGGCCCCGCGACT CGTGCGCCGGGCCGCCGGAGAGGGTGTACTTGGTTCTGAGGCTGTGGTTTCTCCTCC CGCTGAGACCGGTGATCCGCCACCATGGATCTGGAAGCCGCCAAGAATGGCACCGCT TGGA GGCCTA CA TCTGCCGA GGGCGA TTTTGA GCTGGGCA TCA GCA GCAAGCAGAAG CGGAAAAA GA CCAA GA CCGTGAA GA TGA TCGGCGTGCTGA CCCTGTTCCGGTA CA GC GA CTGGCA GGA CAAGCTGTTCA TGA GCCTGGGCA CCA TCA TGGCTA TCGCCCACGGA TCTGGA CTGCCCCTGA TGA TGA TCGTGTTCGGCGA GA TGA CCGA CAAGTTCGTGGA C ACCGCCGGCAACTTCAGCTTCCCCGTGAATTTCAGCCTGAGCCTGCTGAACCCCGGC AAGA TCCTGGAAGAGGAAA TGA CCAGA TA CGCCTACTA CTACAGCGGCCTTGGAGCT GGCGTTCTGGTGGCCGCCTATATCCAGGTGTCCTTTTGGACACTGGCCGCTGGCCGG CA GA TCA GAAA GA TCCGGCAGAAGTTCTTCCA CGCCA TCCTGCGGCAA GA GA TCGGA TGGTTCGA CA TCA A CGA CA CCACCGA GCTGAA CA CCA GA CTGA CCGA CGA CA TCA GC A A GA TCA GCGAAGGCA TCGGCGA CAAA GTGGGAATGTTCTTCCA GGCCGTGGCCA CC TTCTTCGCCGGCTTTA TCGTGGGCTTCA TCA GA GGCTGGAA GCTGA CCCTCGTGA TCA TGGCCA TCTCTCCCA TCCTGGGA CTGTCTGCTGCTGTGTGGGCCAA GA TTCTGA GCGC CTTCA GCGA CAAA GA GCTGGCCGCTTA CGCTAAAGCTGGCGCCGTTGCTGAA GAAGC CCTGGGA GCCA TCA GAA CCGTGA TCGCCTTTGGCGGCCA GA A CA A A GA A CTGGA A A G A TA CCA GAAGCACCTCGA GAA CGCCAAA GA GA TTGGCA TCA A GAA GGCCA TCA GCG CCAA CA TCTCTA TGGGAATCGCCTTCCTGCTGA TCTA CGCCA GCTACGCCCTGGCCTT TTGGTA CGGCTCTA CCCTGGTCA TCAGCAAAGA GT A CA CCA TCGGCAA CGCCA TGA C CGTGTTCTTCTCCA TCCTGA TCGGCGCCTTTA GCGTCGGA CA GGCCGCTCCTTGTA TC GA CGCCTTTGCTAA TGCCA GA GGCGCCGCA TA CGTGA TCTTCGA TA TCA TCGA CAA CA A CCCCAA GA TCGA CA GCTTCA GCGAGCGGGGCCA CAA GCCTGA TA GCA TCA A GGGC AACCTGGAATTCAACGACGTGCACTTCAGCTACCCTAGCCGGGCCAACGTGAAGATC CTGAAGGGCCTGAACCTGAAGGTGCAGAGCGGACAGACAGTGGCCCTCGTGGGATC TTCTGGCTGTGGCAA GA GCACCA CCGTGCAGCTGA TCCAGAGA CTGTA CGA CCCTGA CGA GGGC A CAA TCA A CA TCGA CGGCCA GGA CA TCCGGAA CTTCA A CGTGA ACTA CCT GCGCGA GA TCA TCGGA GTGGTGTCCCA A GA GCCTGTGCTGTTCA GCA CCA CAA TCGC CGA GA AC A TCTGCTACGGCA GA GGCAA CGTGA CCA TGGA CGA GA TCA A GAAA GCCG TGAAA GA GGCCAA CGCCTACGA GTTCA TCA TGAAGCTGCCCCA GAAA TTCGA CA CCC TTGTGGGA GAAA GAGGCGCCCAA CTGTCTGGCGGA CA GA AGCAGAGA ATCGCCA TT GCCAGAGCACTCGTGCGGAACCCTAAGATCCTGCTGCTGGACGAAGCCACAAGCGC CCTGGA TA CA GA GTCTGAGGCAGAAGTGCAGGCTGCCCTGGA CAAGGCTAGAGAGG GCAGAACCA CCATCGTGA TTGCCCACAGA CTGTCCA CCGTGCGGAA TGCCGA TGTGA TCGCCGGA TTCGA A GA TGGCGTGA TCGTGGAA CA GGGCTCCCACAGCGA GCTGA TGA A GAAA GAAGGCGTGTA CTTCA A GCTGGTCAA CA TGC A GA CCA GCGGCTCTCAGA TCC A GA GCGA GGAATTCGA GCTGAA TGA CGA GAA GGCCGCCA CCA GGA TGGCCCCTAA T GGCTGGAAGTCCCGGCTGTTCA GA CA CA GCACCCAGA AGA ATCTGAA GAACAGCCA GA TGTGCCA GAAGTCCCTGGA CGTCGA GA CA GA CGGA CTGGAAGCCAA TGTGCCTCC TGTGTCCTTCCTGAAAGTGCTGAAA CTGAA CAAGA CCGA GTGGCCCTACTTCGTCGTG GGCACCGTGTGTGCCA TTGCTAA TGGTGGA CTGCA GCCCGCCTTCTCCGTGA TCTTTA GCGA GA TTA TCGCCA TCTTTGGCCCTGGCGA CGA CGCCGTGAAGCA GCA GAAGTGTA A TA TCTTTA GCCTGA TCTTCCTGTTCCTGGGGA TTA TCA GCTTCTTTACA TTCTTCCTGC A A GGCTTCA CCTTCGGCAA GGCCGGGGA GA TCCTGACTCGGA GA CTGA GA TCC A TGG CCTTCAAA GCCA TGCTGA GA CA GGA CA TGAGTTGGTTCGA CGA CCA CAA GAA CA GCA CTGGCGCCCTGTCTA CCA GA CTGGCCA CA GA TGCTGCTCAGGTGCAAGGCGCTACAG GCACAA GA CTGGCCCTGA TCGCCCA GAA TA TCGCCA A TCTCGGCA CCGGCA TCA TCATTA GCTTCA TCTA CGGA TGGCA GCTGA CA CTGCTCCTGCTGGCCGTGGTGCCTA TCA T TGCCGTGTCCGGCATCGTGGAAATGAAGTTGCTGGCCGGAAACGCCAAGCGGGACA AGAAAGAACTTGA GGCCGCAGGCAA GA TCGCCACCGA GGCCA TCGAGAA TA TCCGG ACCGTGGTGTCTCTGACCCAAGAGCGGAAGTTCGAGTCTATGTACGTGGAAAAGCTG TACGGCCCCTACAGAAA CA GCGTGCAGAAGGCCCACA TCTACGGCATCA CCTTCA GC ATCAGCCAGGCCTTCATGTACTTTAGCTACGCCGGCTGCTTTAGATTCGGCGCCTACC TGA TCGTGAA CGGCCA CA TGA GA TTCCGGGA CGTGA TCCTGGTGTTCTCCGCCA TCGT GTTTGGAGCCGTGGCTCTGGGACATGCCTCTAGCTTTGCCCCTGACTACGCCAAGGC CAA GCTGTCTGCCGCA CA CCTGTTCA TGCTGTTCGAGA GA CA GCCCCTGA TCGA CTCC TA CA GCGA GGAA GGCCTGAA GCCA GA TAAGTTCGA GGGCAA CA TCA CCTTTAA CGA G GTGGTGTTCAACTA CCCCACCA GA GCCAACGTGCCAGTTCTGCAGGGACTGA GCCTG GAA GTGAA GAA GGGCCA GA CTCTGGCTCTCGTGGGCTCTA GCGGA TGTGGCAA GTCT A CA GTGGTGCA GCTGCTGGAA CGGTTCTACGA TCCTCTGGCTGGAA CCGTCCTGCTG GA TGGCCAA GAA GCCAAAAA GCTGAACGTGCA GTGGCTGA GGGCTCA GCTGGGAA T CGTGTCTCAAGAACCCA TCCTGTTCGA CTGCA GCA TTGCCGA GAA TA TTGCCTA TGGC GA CAA CA GCAGA GTCGTGTCCCA GGA CGA GA TTGTGTCCGCTGCCAAGGCTGCCAAC A TTCA CCCCTTCA TCGA GA CA CTGCCCCA CAAATACGA GA CAAGA GTGGGCGA CAAG GGCACACAGCTGAGCGGAGGCCA GAAA CA GA GGA TTGCTA TCGCTCGCGCCCTGA T CA GA CA GCCTCA GA TTCTGCTCCTGGA TGAGGCCA CCTCTGCTCTGGA CA CCGA GA G CGA GAAGGTGGTGCAA GA GGCTCTGGA TAA GGCCCGCGA GGGAA GAA CCTGTA TCG TGA TCGCTCACCGGCTGA GCA CCA TCCA GAA TGCCGA CCTCA TCGTGGTGTTCCAGA A CGGCA GAGTGAAAGA GCA CGGCA CCCA CCA GCA A CTGCTGGCCCA GAAGGGCATC TACTTCTCCATGGTGTCTGTGCAGGCCGGCACACAGAACCTGTGAGTCGACAATCAAC CTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTTAC GCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCTTT CATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCCGT TGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTTGG GGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGC CACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTGTT GGGCACTGACAATTCCGTGGTGTTGTCGGGGAAGCTGACGTCCTTTCCATGGCTGCTC GCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCCC TCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGCG TCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCCTGGA ATTCGAGCTCGGTACCTTTAAGACCAATGACTTACAAGGCAGCTGTAGATCTTAGCCAC TTTTTAAAAGAAAAGGGGGGACTGGAAGGGCTAATTCACTCCCAACGAAGACAAGATCT GCTTTTTGCTTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTG GCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTA GTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTC AGTGTGGAAAATCTCTAGCAGTAGTAGTTCATGTCATCTTATTATTCAGTATTTATAACTT GCAAAGAAATGAATATCAGAGAGTGAGAGGAACTTGTTTATTGCAGCTTATAATGGTTAC AAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTT GTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGCTCTAGCTATCCCGCCCCT AACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATGGC TGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTATTCCA GAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGGACGTACCCAATTCGCCCTATAGTGA GTCGTATTACGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTG GCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGC GAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGG GACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTG ACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCT CGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTC CGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACG TAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCT TTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTT TTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTTAGGTGGCACTTTTCGGG GAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCT CATGAGACAATAACCCTGATAAATGCTTCAATAATAGCACCTAGATCAAGAGACAGGATG AGGATCGTTTCGCATGATTGAACAAGATGGATTGCACGCAGGTTCTCCGGCCGCTTGG GTGGAGAGGCTATTCGGCTATGACTGGGCACAACAGACAATCGGCTGCTCTGATGCCG CCGTGTTCCGGCTGTCAGCGCAGGGGCGCCCGGTTCTTTTTGTCAAGACCGACCTGTC CGGTGCCCTGAATGAACTGCAAGACGAGGCAGCGCGGCTATCGTGGCTGGCCACGAC GGGCGTTCCTTGCGCAGCTGTGCTCGACGTTGTCACTGAAGCGGGAAGGGACTGGCT GCTATTGGGCGAAGTGCCGGGGCAGGATCTCCTGTCATCTCACCTTGCTCCTGCCGAG AAAGTATCCATCATGGCTGATGCAATGCGGCGGCTGCATACGCTTGATCCGGCTACCTG CCCATTCGACCACCAAGCGAAACATCGCATCGAGCGAGCACGTACTCGGATGGAAGCC GGTCTTGTCGATCAGGATGATCTGGACGAAGAGCATCAGGGGCTCGCGCCAGCCGAAC TGTTCGCCAGGCTCAAGGCGAGCATGCCCGACGGCGAGGATCTCGTCGTGACCCATG GCGATGCCTGCTTGCCGAATATCATGGTGGAAAATGGCCGCTTTTCTGGATTCATCGAC TGTGGCCGGCTGGGTGTGGCGGACCGCTATCAGGACATAGCGTTGGCTACCCGTGATA TTGCTGAAGAGCTTGGCGGCGAATGGGCTGACCGCTTCCTCGTGCTTTACGGTATCGC CGCTCCCGATTCGCAGCGCATCGCCTTCTATCGCCTTCTTGACGAGTTCTTCTGAATTAT TAACGCTTACAATTTCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACA CCGCATCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTC TAAATACATTCAAATATGTATCCGCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCC ACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGC GCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCG GATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACC AAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACC GCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGT CGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGG CTGAACGGGGGGGTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCG GACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCA GGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCG TCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCG GCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTA TCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCG CAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAAT ACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGG TTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCA TTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGA GCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCGCGCAATT AACCCTCACTAAAGGGAACAAAAGCTGGAGCTGCAAGCTTGGCCATTGCATACGTTGTA TCCATATCATAATATGTACATTTATATTGGCTCATGTCCAACATTACCGCCATGTTGACAT TGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATTAGTTCATAGCCCATATA TGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTGGCTGACCGCCCAACGA CCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTAACGCCAATAGGGACTT TCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCACTTGGCAGTACATCAA GTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACGGTAAATGGCCCGCCTG GCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGGCAGTACATCTACGTATT AGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATCAATGGGCGTGGATAGC GGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGTCAATGGGAGTTTGTTT TGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTCCGCCCCATTGACGCA AATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGAGCTCGTTTAGTGAAC CGGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGGCTAACTAGGGAA CCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCCCGTC TGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAAAATCT CTAGCAGTGGCGCCCGAACAGGGACTTGAAAGCGAAAGGGAAACCAGAGGAGCTCTCT CGACGCAGGACTCGGCTTGCTGAAGCGCGCACGGCAAGAGGCGAGGGGCGGCGACTGGTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGAAGGAGAGAGATGGGTGCGAG AGCGTCAGTATTAAGCGGGGGAGAATTAGATCGCGATGGGAAAAAATTCGGTTAAGGC CAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTATGGGCAAGCAGGGAGCTAGAA CGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGTAGACAAATACTGGG ACAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATCATTATATAATACAGT AGCAACCCTCTATTGTGTGCATCAAAGGATAGAGATAAAAGACACCAAGGAAGCTTTAG ACAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCACAGCAAGCGGCCGCTGA TCTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAGTGAATTATATAAATAT AAAGTAGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGCAAAGAGAAGAGTGGT GCAGAGAGAAAAAAGAGCAGTGGGAATAGGAGCTTTGTTCCTTGGGTTCTTGGGAGCA GCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACAGGCCAGACAATTAT TGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTGAGGCGCAACAGCAT CTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAGAATCCTGGCTGTGG AAAGATACCTAAAGGATCAACAGCTCCTGGGGATTTGGGGTTGCTCTGGAAAACTCATT TGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTCTGGAACAGATTTG GAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTACACAAGCTTAATAC ACTCCTTAATTGAAGAATCGCAAAACCAGCAAGAAAAGAATGAACAAGAATTATTGGAAT TAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACATAACAAATTGGCTGTGGTATATAA AATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTTTTGCTGTACTTTC TATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGACCCACCTCCCAAC CCCGAGGGGACCCGACAGGCCCGAAGGAATAGAAGAAGAAGGTGGAGAGAGAGACAG AGACAGATCCATTCGATTAGTGAACGGATCTCGACGGTATCGGTT

[0256] Annotations

[0257] • 4..121: cPPT / CTS_ central polypurine tract and central termination sequence of HIV-1 • 139..972: LPE _LPa promoter combined with 287bp enhancer region found upstream natural ABCB4 promoter (shown bold)

[0258] • 990..4829: coABCB4_ codon optimised version of the human ABCB4 gene (shown bold and in italics)

[0259] • 4836..5424: WPRE_ woodchuck hepatitis virus posttranscriptional regulatory element • 5511..5744: 3' LTR (AU3)_ self-inactivating 3' long terminal repeat (LTR) from HIV-1 • 5816..5937: SV40 poly(A) signal_ SV40 polyadenylation signal

[0260] • 5938..9075: Part of the plasmid needed for bacterial work

[0261] • 9076..9659: CMV promoter - allows expression of the vector genome

[0262] • 9659..5744: Vector genome

[0263] • 9674..9854: 5' LTR (truncated)- truncated 5' long terminal repeat (LTR) from HIV-1 • 9901..10026: HIV-1 packaging signal of human immunodeficiency virus type 1 • 10519..10752: RRE_ The Rev response element (RRE) of HIV-1 allows for Rev- dependent mRNA export from the nucleus to the cytoplasm

[0264] SEQ ID NO: 31 - CCL. LPa. Luc. GFP, Version 2 backbone - GFP CCATTGCATACGTTGTATCCATATCATAATATGTACATTTATATTGGCTCATGTCCAACAT TACCGCCATGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATT AGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTG GCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTA ACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCA CTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACG GTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGG CAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATC AATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGT CAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTC CGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGA GCTCGTTTAGTGAACCGGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCT GGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGT AGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAAAATCTCTAGCAGTGGCGCCCGAACAGGGACTTGAAAGCGAAAGGGAAA CCAGAGGAGCTCTCTCGACGCAGGACTCGGCTTGCTGAAGCGCGCACGGCAAGAGGC GAGGGGCGGCGACTGGTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGAAGGAGA GAGATGGGTGCGAGAGCGTCAGTATTAAGCGGGGGAGAATTAGATCGCGATGGGAAAA AATTCGGTTAAGGCCAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTATGGGCAA GCAGGGAGCTAGAACGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGT AGACAAATACTGGGACAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATC ATTATATAATACAGTAGCAACCCTCTATTGTGTGCATCAAAGGATAGAGATAAAAGACAC CAAGGAAGCTTTAGACAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCACAG CAAGCGGCCGCTGATCTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAGT GAATTATATAAATATAAAGTAGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGCA AAGAGAAGAGTGGTGCAGAGAGAAAAAAGAGCAGTGGGAATAGGAGCTTTGTTCCTTG GGTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACA GGCCAGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTG AGGCGCAACAGCATCTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAG AATCCTGGCTGTGGAAAGATACCTAAAGGATCAACAGCTCCTGGGGATTTGGGGTTGCT CTGGAAAACTCATTTGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTC TGGAACAGATTTGGAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTAC ACAAGCTTAATACACTCCTTAATTGAAGAATCGCAAAACCAGCAAGAAAAGAATGAACAA GAATTATTGGAATTAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACATAACAAATTGG CTGTGGTATATAAAATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTT TTGCTGTACTTTCTATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGA CCCACCTCCCAACCCCGAGGGGACCCGACAGGCCCGAAGGAATAGAAGAAGAAGGTG GAGAGAGAGACAGAGACAGATCCATTCGATTAGTGAACGGATCTCGACGGTATCGGTTA ACTTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGACA TAATAGCAACAGACATACAAACTAAAGAATTACAAAAACAAATTACAAAAATTCAAAATTT TATCGATCACGAGACTAGCCTCGAGGGGGTGGCCCATCAGGCGAGGCTTTCTCGGCA CCCGAGGCTCCAGCCTGATCTCGGTCTTATCCTGCGACCGGGCTGGTTCTGGCGGGT CGCCAGGGTGGGCGGCGGCCCCAGCCGGGCGCCCCGGCGGCAAGAGCGGCAGGCT GCGCCCCTGGCCCGCGCCTAGCCTGGGGAGAGAGCTGGGCGGGCGGCGGGAGCTG CTCTCGCGGGCCGCGGCCCTCGCCCTGGCTGCAACGGTAGGCGTTTCCCGGGCCGG ACGCGCGTGGGGGGCGGGGGCGGGGGCGGGGGCGAGGCCGCGGCGAGCAAAGTC CAGGCCCCTCTGCTGCAGCGCCCGCGCGTCCAGAGGCCCTGCCAGACACGCGCGAG GTTCGAGCCGAGAGAGGTCCGGGCGCGTCTGGCCTCGAAGGGAGACCCGGGACGTG GGGCGCGGGGCGGGAGTGGCCGGACCTCCACCCAGTGCCCCCGGGCCCCGCGACT CGTGCGCCGGGCCGCCGGAGAGGGTGTACTTGGTTCTGAGGCTGTGGTTTCTCCTCC CGCTGAG ACCGGTGCCACCATG GA CT A CA A A GA CCA TGA CGGTGA TTA TA A AG A TCA TGA CA TCGA TTACAA GGA TGA CGA TGA CAAGA GCGGCGGCGGCAGCA TGGA GGA CG CCAAGAACATCAAGAAGGGACCAGCCCCCAGATACCCCCTGGAGGACGGCACAGCC GGCGA GCAGCTGCACAA GGCCA TGA A GCGGTA CGCCCA GGTGCCA GGCA CCA TCGC CTTCA CCGA CGCCCA CA TCGA GGTGAACA TCA CCTA CGCCGA GT A CTTCGA GA TGA G CGTGCGGCTGGCCGAGGCCATGAAGCGGTACGGCCTGAACACCAACCACCGGATCG TGGTGTGCAGCGA GAA CA GCCTGCA GTTCTTCA TGCCCGTGCTGGGA GCCCTGTTCA T CGGCGTGGCCGTGGCCCCAGCCAACGACATCTACAACGAGCGGGAGCTGCTGAACA GCA TGAACA TCAGCCA GCCCACCGTGGTGTTCGTGA GCA A GAA GGGCCTGCA GA AG A TCCTGAA TGTGCA GAA GAA GCTGCCCA TCA TCCA GAA GA TCA TCA TCA TGGA CA GC A AG A CCGA TTA CCAGGGCTTCCA GA GCA TGTA CACCTTCGTGA CCA GCCA CCTGCCC CCA GGCTTCAACGA GT A CGA CTTCA A GCCCGA GA GCTTCGA CCGGGA CAA GA CCA TC GCCCTGA TCA TGAACA GCAGCGGCA GCA CCGGCCTGCCCAA GGGCGTGGCCCTGCC CCA CCGGA CCGCCTGCGTGCGGTTCA GCCA CGCCA GA GA CCCCA TCTTCGGCAA CCA GA TCA A GCCCGA CA CCGCCA TCCTGA GCGTGGTGCCCTTCCACCA CGGCTTCGGCA T GTTCA CCACCCTGGGCTA CCTGA TCTGCGGCTTCCGGGTGGTGCTGA TGTA CA GGTTC GA GGA GGA GCTGTTCCTGCGGA GCCTGCAGGA CT AC A A GA TCCA GA GCGCCCTGCT GGTGCCCA CCCTGTTCA GCTTCTTCGCCAA GA GCA CCCTGA TCGACAA GT A CGA CCT GA GCA A CCTGCACGA GA TCGCCTCTGGCGGA GCCCCA CTGA GCAAGGAGGTGGGCGA GGCCGTGGCCAAGCGGTTCCA CCTGCCA GGCA TCCGGCAGGGCTA CGGCCTGA CC GA GA CCACCA GCGCCA TCCTGA TCA CCCCCGA GGGCGA CGA CAAGCCCGGA GCCGT GGGCAAGGTGGTGCCCTTCTTCGAGGCCAAGGTGGTGGACCTGGACACCGGCAAGA CCCTGGGCGTGAACCA GA GA GGCGA GCTGTGCGTGA GAGGCCCCA TGA TCA TGA GC GGCTA CGTGAA CAA CCCCGA GGCCA CCA A CGCCCTGA TCGA CAAGGA CGGCTGGCT GCA CA GCGGCGA CA TCGCCT ACTGGGA CGAGGA CGA GCA CTTCTTCA TCGTGGA CCG GCTGA AG A GCCTGA TCA A GTACAAGGGCTA CCA GGTGGCCCCA GCCGA GCTGGA GA GCA TCCTGCTGCAGCA CCCCAA CA TCCGGGA CGCCGGAGTGGCCGGA CTGCCCGA C GA CGA CGCCGGA GA GCTGCCA GCCGCCGTGGTGGTGCTGGA GCACGGCAA GA CCA T GA CCG A GAAGGA GA TCGTGGA CTACGTGGCCA GCCA GG TGA CCA CCGCCAA GAAGC TGAGAGGCGGCGTGGTGTTCGTGGACGAGGTGCCCAAGGGCCTGACCGGCAAGCTG GA CGCCA GAAA GA TCCGGGA GA TCCTGA TCA A GGCCA A GAA GGGCGGCAA GA TCCG CGCAGAGGGCCGGGGCTCATTGCTGACCTGTGGAGATGTCGAGGAAAATCCCGGCC CCATGGTGAGCAAGGGCGAGGAGCTGTTCACCGGGGTGGTGCCCATCCTGGTCGAG CTGGA CGGCGA CGTAAA CGGCCA CAAGTTCA GCGTGTCCGGCGA GGGCGA GGGCGA TGCCA CCTACGGCAA GCTGA CCCTGAA GTTCA TCTGCA CCA CCGGCAA GCTGCCCGT GCCCTGGCCCACCCTCGTGA CCA CCCTGA CCTACGGCGTGCAGTGCTTCA GCCGCTA CCCCGA CCACA TGAAGCA GCA CGA CTTCTTCAAGTCCGCCA TGCCCGAA GGCTA CGT CCA GGA GCGCA CCA TCTTCTTCAA GGA CGA CGGCAACTACAA GA CCCGCGCCGAGGT GAA GTTCGA GGGCGA CACCCTGGTGAA CCGCA TCGA GCTGA A GGGCA TCGA CTTCAA GGAGGA CGGCAACA TCCTGGGGCACAA GCTGGAGTACAA CTACAA CAGCCACAACG TCTA TA TCA TGGCCGA CAA GCA GAA GAA CGGCA TCAAGGTGAA CTTCAA GA TCCGCC A CAA CA TCGA GGA CGGCA GCGTGCAGCTCGCCGA CCACTA CCA GCA GAA CA CCCCC A TCGGCGA CGGCCCCGTGCTGCTGCCCGA CAACCA CT A CCTGA GCA CCCAGTCCGCC CTGA GCAAAGA CCCCAA CGA GAAGCGCGA TCA CA TGGTCCTGCTGGA GTTCGTGA CC GCCGCCGGGA TCACTCTCGGCA TGGA CGAGCTGTA CAA GTAAGTCGGTCGACAATCA ACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAACTATGTTGCTCCTTTT ACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATTGCTTCCCGTATGGCT TTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATGAGGAGTTGTGGCCC GTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGCAACCCCCACTGGTT GGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATT GCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGACAGGGGCTCGGCTG TTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAGCTGACGTCCTTTCCATGGCTGCT CGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTACGTCCCTTCGGCC CTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTGCGGCCTCTTCCGC GTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCGCCTCCCCGCCTGG AATTCGAGCTCGGTACCTTTAAGACCAATGACTTACAAGGCAGCTGTAGATCTTAGCCA CTTTTTAAAAGAAAAGGGGGGACTGGAAGGGCTAATTCACTCCCAACGAAGACAAGATC TGCTTTTTGCTTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCT GGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGT AGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGT CAGTGTGGAAAATCTCTAGCAGTAGTAGTTCATGTCATCTTATTATTCAGTATTTATAACT TGCAAAGAAATGAATATCAGAGAGTGAGAGGAACTTGTTTATTGCAGCTTATAATGGTTA CAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGT TGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGCTCTAGCTATCCCGCCCC TAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATGG CTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTATTCCA GAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGGACGTACCCAATTCGCCCTATAGTGA GTCGTATTACGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTG GCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGC GAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGG GACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTG ACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCT CGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTC CGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCT TTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTT TTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACA AAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTTAGGTGGCACTTTTCGGG GAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCT CATGAGACAATAACCCTGATAAATGCTTCAATAATAGCACCTAGATCAAGAGACAGGATG AGGATCGTTTCGCATGATTGAACAAGATGGATTGCACGCAGGTTCTCCGGCCGCTTGG GTGGAGAGGCTATTCGGCTATGACTGGGCACAACAGACAATCGGCTGCTCTGATGCCG CCGTGTTCCGGCTGTCAGCGCAGGGGCGCCCGGTTCTTTTTGTCAAGACCGACCTGTC CGGTGCCCTGAATGAACTGCAAGACGAGGCAGCGCGGCTATCGTGGCTGGCCACGAC GGGCGTTCCTTGCGCAGCTGTGCTCGACGTTGTCACTGAAGCGGGAAGGGACTGGCT GCTATTGGGCGAAGTGCCGGGGCAGGATCTCCTGTCATCTCACCTTGCTCCTGCCGAG AAAGTATCCATCATGGCTGATGCAATGCGGCGGCTGCATACGCTTGATCCGGCTACCTG CCCATTCGACCACCAAGCGAAACATCGCATCGAGCGAGCACGTACTCGGATGGAAGCC GGTCTTGTCGATCAGGATGATCTGGACGAAGAGCATCAGGGGCTCGCGCCAGCCGAAC TGTTCGCCAGGCTCAAGGCGAGCATGCCCGACGGCGAGGATCTCGTCGTGACCCATG GCGATGCCTGCTTGCCGAATATCATGGTGGAAAATGGCCGCTTTTCTGGATTCATCGAC TGTGGCCGGCTGGGTGTGGCGGACCGCTATCAGGACATAGCGTTGGCTACCCGTGATA TTGCTGAAGAGCTTGGCGGCGAATGGGCTGACCGCTTCCTCGTGCTTTACGGTATCGC CGCTCCCGATTCGCAGCGCATCGCCTTCTATCGCCTTCTTGACGAGTTCTTCTGAATTAT TAACGCTTACAATTTCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACA CCGCATCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTC TAAATACATTCAAATATGTATCCGCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCC ACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGC GCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCG GATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACC AAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACC GCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGT CGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGG CTGAACGGGGGGGTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACT GAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCG GACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCA GGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCG TCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCG GCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTA TCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCG CAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAAT ACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGG TTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCA TTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGA GCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCGCGCAATT AACCCTCACTAAAGGGAACAAAAGCTGGAGCTGCAAGCTTGG

[0265] Annotations

[0266] • 74..657: CMV promoter - allows expression of the vector genome

[0267] • 658..6366: Vector genome

[0268] • 672..852: 5' LTR (truncated)- truncated 5' long terminal repeat (LTR) from HIV-1

[0269] • 899..1024: HIV-1 Ψ_ packaging signal of human immunodeficiency virus type 1

[0270] • 1517..1750: RRE_ The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm

[0271] • 2246..2363: cPPT / CTS_ central polypurine tract and central termination sequence of HIV- 1

[0272] • 2389..2927: LPa promoter_ novel promoter (shown bold)

[0273] • 2943..5447: Transgene (shown bold and in italics)• 2943..3008: three tandem FLAG® epitope tags, followed by an enterokinase cleavage site (shown bold and in italics)

[0274] • 3009..4673: Luciferase (shown bold and in italics)

[0275] • 6474..4727: 2A peptide from Thosea asigna virus capsid protein (shown bold and in italics) • 4728..5447: EGFP_ mammalian codon-optimized enhanced GFP (shown bold and in italics)

[0276] • 5458..6046: WPRE_ woodchuck hepatitis virus posttranscriptional regulatory element • 6133..6366: 3' LTR (AU3)_ self-inactivating 3' long terminal repeat (LTR) from HIV-1 • 6438..6559: SV40 poly(A) signal_ SV40 polyadenylation signal

[0277] • 6560..74: Part of the plasmid needed for bacterial work

[0278] SEQ ID NO: 32 - miR-G-LPaH, Version 3 backbone - GFP CCATTGCATACGTTGTATCCATATCATAATATGTACATTTATATTGGCTCATGTCCAACAT TACCGCCATGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATT AGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTG GCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTA ACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCA CTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACG GTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGG CAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATC AATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGT CAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTC CGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGA GCTCGTTTAGTGAACCGGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCT GGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGT AGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGT CAGTGTGGAAAATCTCTAGCAGTGGCGCCCGAACAGGGACTTGAAAGCGAAAGGGAAA CCAGAGGAGCTCTCTCGACGCAGGACTCGGCTTGCTGAAGCGCGCACGGCAAGAGGC GAGGGGCGGCGACTGGTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGAAGGAGA GAGATGGGTGCGAGAGCGTCAGTATTAAGCGGGGGAGAATTAGATCGCGATGGGAAAA AATTCGGTTAAGGCCAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTATGGGCAA GCAGGGAGCTAGAACGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGT AGACAAATACTGGGACAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATC ATTATATAATACAGTAGCAACCCTCTATTGTGTGCATCAAAGGATAGAGATAAAAGACAC CAAGGAAGCTTTAGACAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCACAG CAAGCGGCCGCTGATCTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAGT GAATTATATAAATATAAAGTAGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGCA AAGAGAAGAGTGGTGCAGAGAGAAAAAAGAGCAGTGGGAATAGGAGCTTTGTTCCTTG GGTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACA GGCCAGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTG AGGCGCAACAGCATCTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAG AATCCTGGCTGTGGAAAGATACCTAAAGGATCAACAGCTCCTGGGGATTTGGGGTTGCT CTGGAAAACTCATTTGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTC TGGAACAGATTTGGAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTAC ACAAGCTTAATACACTCCTTAATTGAAGAATCGCAAAACCAGCAAGAAAAGAATGAACAA GAATTATTGGAATTAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACATAACAAATTGG CTGTGGTATATAAAATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTT TTGCTGTACTTTCTATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGA CCCACCTCCCAACCCCGAGGGGACCCGACAGGCCCGAAGGAATAGAAGAAGAAGGTG GAGAGAGAGACAGAGACAGATCCATTCGATTAGTGAACGGATCTCGACGGTATCGGTTA ACTTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGACA TAATAGCAACAGACATACAAACTAAAGAATTACAAAAACAAATTACAAAAATTCAAAATTT TATCGATCACGAGACTAGCCTCGAGGATATCCCATAGAGCCCACCGCATCCCCAGCAT GCCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCAGAATAGAATGACACCTACTCAGACAATGCGATGCAATTTCCTCATTTTATTAGGAAAGGACA GTGGGAGTGGCACCTTCCAGGGTCAAGGAAGGCACGGGGGAGGGGCAAACAACAGAT GGCTGGCAACTAGAAGGCACAGACGCGTGGGCCCCTGCAGTCGTACCGTGAGTAATAA TGCGATCGTCGTACCGTGAGTAATAATGCGCGGTTCGTACCGTGAGTAATAATGCGATC GTCGTACCGTGAGTAATAATGCGCTGCAGGTGTAGTGTTTCCTACTTTATGGAGTGATG TAGTGTTTCCTACTTTATGGACGGTTGTAGTGTTTCCTACTTTATGGAATCGTGTAGTGTT TCCTACTTTATGGAGGCTAGCCTAGGCGGGGAGGCGGCCCAAAGGGAGATCCGACTC GTCTGAGGGCGAAGGCGAAGACGCGGAAGAGGCCGCAGAGCCGGCAGCAGGCCGCG GGAAGGAAGGTCCGCTGGATTGAGGGCCGAAGGGACGTAGCAGAAGGACGTCCCGCG CAGAATCCAGGTGGCAACACAGGCGAGCAGCCATGGAAAGGACGTCAGCTTCCCCGAC AACACCACGGAATTGTCAGTGCCCAACAGCCGAGCCCCTGTCCAGCAGCGGGCAAGG CAGGCGGCGATGAGTTCCGCCGTGGCAATAGGGAGGGGGAAAGCGAAAGTCCCGGAA AGGAGCTGACAGGTGGTGGCAATGCCCCAACCAGTGGGGGTTGCGTCAGCAAACACA GTGCACACCACGCCACGTTGCCTGACAACGGGCCACAACTCCTCATAAAGAGACAGCA ACCAGGATTTATACAAGGAGGAGAAAATGAAAGCCATACGGGAAGCAATAGCATGATAC AAAGGCATTAAAGCAGCGTATCCACATAGCGTAAAAGGAGCAACATAGTTAAGAATACC AGTCAATCTTTCACAAATTTTGTAATCCAGAGGTTGATTGTCGACGCGGCCGCT7TAC7T GT A CA GCTCGTCCA TGCCGA GAGTGA TCCCGGCGGCGGTCA CGAA CTCCA GCA GGA CCATGTGATCGCGCTTCTCGTTGGGGTCTTTGCTCAGGGCGGACTGGGTGCTCAGGTA GTGGTTGTCGGGCAGCAGCACGGGGCCGTCGCCGATGGGGGTGTTCTGCTGGTAGTG GTCGGCGAGCTGCACGCTGCCGTCCTCGA TGTTGTGGCGGA TCTTGAAGTTCA CCTTG A TGCCGTTCTTCTGCTTGTCGGCCA TGA TA TA GA CGTTGTGGCTGTTGTA GTTGTA CTC CA GCTTGTGCCCCA GGA TGTTGCCGTCCTCCTTGA A GTCGA TGCCCTTCA GCTCGA TG CGGTTCACCAGGGTGTCGCCCTCGAACTTCACCTCGGCGCGGGTCTTGTAGTTGCCGT CGTCCTTGAAGAAGATGGTGCGCTCCTGGACGTAGCCTTCGGGCATGGCGGACTTGA AGAAGTCGTGCTGCTTCATGTGGTCGGGGTAGCGGCTGAAGCACTGCACGCCGTAGG TCAGGGTGGTCACGAGGGTGGGCCAGGGCACGGGCAGCTTGCCGGTGGTGCAGATG AACTTCAGGGTCAGCTTGCCGTAGGTGGCATCGCCCTCGCCCTCGCCGGACACGCTG AACTTGTGGCCGTTTACGTCGCCGTCCAGCTCGACCAGGATGGGCACCACCCCGGTG AACAGCTCCTCGCCCTTGCTCACCATGGTGGCGACCGGTCTCAGCGGGAGGAGAAAC CACAGCCTCAGAACCAAGTACACCCTCTCCGGCGGCCCGGCGCACGAGTCGCGGGG CCCGGGGGCACTGGGTGGAGGTCCGGCCACTCCCGCCCCGCGCCCCACGTCCCGGG TCTCCCTTCGAGGCCAGACGCGCCCGGACCTCTCTCGGCTCGAACCTCGCGCGTGTC TGGCAGGGCCTCTGGACGCGCGGGCGCTGCAGCAGAGGGGCCTGGACTTTGCTCGC CGCGGCCTCGCCCCCGCCCCCGCCCCCGCCCCCCACGCGCGTCCGGCCCGGGAAAC GCCTACCGTTGCAGCCAGGGCGAGGGCCGCGGCCCGCGAGAGCAGCTCCCGCCGCC CGCCCAGCTCTCTCCCCAGGCTAGGCGCGGGCCAGGGGCGCAGCCTGCCGCTCTTG CCGCCGGGGCGCCCGGCTGGGGCCGCCGCCCACCCTGGCGACCCGCCAGAACCAG CCCGGTCGCAGGATAAGACCGAGATCAGGCTGGAGCCTCGGGTGCCGAGAAAGCCT CGCCTGATGGGCCACCCCCTCGAGGCCTCTCACACTACCTAAACCACGCCAGGACAA CCTCTGCTCCTCTCCA CCGAAA TTCCAA GGGGTCGAGTGGA TGTTGGA GGTGGCA TG GGCCCAGAGAGGTCTCTGACCTCTGCCCCAGCTCCAAGGTCAGCAGGCAGGGAGGG CTGTGTGTTTGCTGTTTGCTGCTTGCAA TGTTTGCCCA TTTTA GGGGATATGCTTGAATT CGAGCTCGGTACCTTTAAGACCAATGACTTACAAGGCAGCTGTAGATCTTAGCCACTTTT TAAAAGAAAAGGGGGGACTGGAAGGGCTAATTCACTCCCAACGAAGACAAGATCTGCTT TTTGCTTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGGCTA ACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGT GTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTG TGGAAAATCTCTAGCAGTAGTAGTTCATGTCATCTTATTATTCAGTATTTATAACTTGCAA AGAAATGAATATCAGAGAGTGAGAGGAACTTGTTTATTGCAGCTTATAATGGTTACAAAT AAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAGTTGTGG TTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGCTCTAGCTATCCCGCCCCTAACT CCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATGGCTGAC TAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTATTCCAGAAGT AGTGAGGAGGCTTTTTTGGAGGCCTAGGGACGTACCCAATTCGCCCTATAGTGAGTCGTATTACGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTT ACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGA GGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGGACGC GCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGC TACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCA CGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTT AGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCACGTAGTGG GCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATA GTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTCTTTTGATT TATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAACAAAAATT TAACGCGAATTTTAACAAAATATTAACGCTTACAATTTAGGTGGCACTTTTCGGGGAAAT GTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGA GACAATAACCCTGATAAATGCTTCAATAATAGCACCTAGATCAAGAGACAGGATGAGGA TCGTTTCGCATGATTGAACAAGATGGATTGCACGCAGGTTCTCCGGCCGCTTGGGTGG AGAGGCTATTCGGCTATGACTGGGCACAACAGACAATCGGCTGCTCTGATGCCGCCGT GTTCCGGCTGTCAGCGCAGGGGCGCCCGGTTCTTTTTGTCAAGACCGACCTGTCCGGT GCCCTGAATGAACTGCAAGACGAGGCAGCGCGGCTATCGTGGCTGGCCACGACGGGC GTTCCTTGCGCAGCTGTGCTCGACGTTGTCACTGAAGCGGGAAGGGACTGGCTGCTAT TGGGCGAAGTGCCGGGGCAGGATCTCCTGTCATCTCACCTTGCTCCTGCCGAGAAAGT ATCCATCATGGCTGATGCAATGCGGCGGCTGCATACGCTTGATCCGGCTACCTGCCCA TTCGACCACCAAGCGAAACATCGCATCGAGCGAGCACGTACTCGGATGGAAGCCGGTC TTGTCGATCAGGATGATCTGGACGAAGAGCATCAGGGGCTCGCGCCAGCCGAACTGTT CGCCAGGCTCAAGGCGAGCATGCCCGACGGCGAGGATCTCGTCGTGACCCATGGCGA TGCCTGCTTGCCGAATATCATGGTGGAAAATGGCCGCTTTTCTGGATTCATCGACTGTG GCCGGCTGGGTGTGGCGGACCGCTATCAGGACATAGCGTTGGCTACCCGTGATATTGC TGAAGAGCTTGGCGGCGAATGGGCTGACCGCTTCCTCGTGCTTTACGGTATCGCCGCT CCCGATTCGCAGCGCATCGCCTTCTATCGCCTTCTTGACGAGTTCTTCTGAATTATTAAC GCTTACAATTTCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTTCACACCGC ATCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAAT ACATTCAAATATGTATCCGCTCATGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTG AGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGT AATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATC AAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAAT ACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCC TACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGT GTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTG AACGGGGGGGTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAG ATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGAC AGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGG GGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCG ATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCC TTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCC CCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAG CCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACG CAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTT CCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTA GGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCG GATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCGCGCAATTAAC CCTCACTAAAGGGAACAAAAGCTGGAGCTGCAAGCTTGG

[0279] Annotations:

[0280] • 74..657: CMV promoter - allows expression of the vector genome

[0281] • 658..5452: Vector genome

[0282] • 672..852: 5' LTR (truncated)- truncated 5' long terminal repeat (LTR) from HIV-1

[0283] • 899..1024: HIV-1

[0284]

[0285] packaging signal of human immunodeficiency virus type 11517..1750: RRE_ The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm

[0286] 2246..2363: cPPT / CTS_ central polypurine tract and central termination sequence of HIV- 1

[0287] 2394..2618: bGH poly(A) signal_ bovine growth hormone polyadenylation signal 2637..2736: 4x miR-126-3pT binding site (shown underlined)

[0288] 2744..2847: 4x miR142 binding site (shown underlined)

[0289] 2859..3447: WPRE_ woodchuck hepatitis virus posttranscriptional regulatory element 3463..4182: EGFP_ mammalian codon-optimized enhanced GFP (shown bold and in italics)

[0290] 4196..4734: LPa promoter_ novel promoter (shown bold)

[0291] 4743..4933: HCR_ hApoE-HCR (Apolipoprotein E Hepatic Control Region) (shown bold and underlined)

[0292] 5026..5259: 3' LTR (AU3)_ self-inactivating 3' long terminal repeat (LTR) from HIV-1 5331..5452: SV40 poly(A) signal_ SV40 polyadenylation signal

[0293] 5453..74: Part of the plasmid needed for bacterial work

[0294] SEQ ID NO: 33 - miR-coA4-LPaH, Version 3 backbone - coABCB4 CCATTGCATACGTTGTATCCATATCATAATATGTACATTTATATTGGCTCATGTCCAACAT TACCGCCATGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATT AGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTG GCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTA ACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCA CTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACG GTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGG CAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATC AATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGT CAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTC CGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGA GCTCGTTTAGTGAACCGGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCT GGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGT AGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGT CAGTGTGGAAAATCTCTAGCAGTGGCGCCCGAACAGGGACTTGAAAGCGAAAGGGAAA CCAGAGGAGCTCTCTCGACGCAGGACTCGGCTTGCTGAAGCGCGCACGGCAAGAGGC GAGGGGCGGCGACTGGTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGAAGGAGA GAGATGGGTGCGAGAGCGTCAGTATTAAGCGGGGGAGAATTAGATCGCGATGGGAAAA AATTCGGTTAAGGCCAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTATGGGCAA GCAGGGAGCTAGAACGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGT AGACAAATACTGGGACAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATC ATTATATAATACAGTAGCAACCCTCTATTGTGTGCATCAAAGGATAGAGATAAAAGACAC CAAGGAAGCTTTAGACAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCACAG CAAGCGGCCGCTGATCTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAGT GAATTATATAAATATAAAGTAGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGCA AAGAGAAGAGTGGTGCAGAGAGAAAAAAGAGCAGTGGGAATAGGAGCTTTGTTCCTTG GGTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACA GGCCAGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTG AGGCGCAACAGCATCTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAG AATCCTGGCTGTGGAAAGATACCTAAAGGATCAACAGCTCCTGGGGATTTGGGGTTGCT CTGGAAAACTCATTTGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTC TGGAACAGATTTGGAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTAC ACAAGCTTAATACACTCCTTAATTGAAGAATCGCAAAACCAGCAAGAAAAGAATGAACAA GAATTATTGGAATTAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACATAACAAATTGG CTGTGGTATATAAAATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTT TTGCTGTACTTTCTATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGACCCACCTCCCAACCCCGAGGGGACCCGACAGGCCCGAAGGAATAGAAGAAGAAGGTG GAGAGAGAGACAGAGACAGATCCATTCGATTAGTGAACGGATCTCGACGGTATCGGTTA ACTTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGACA TAATAGCAACAGACATACAAACTAAAGAATTACAAAAACAAATTACAAAAATTCAAAATTT TATCGATCACGAGACTAGCCTCGAGGATATCCCATAGAGCCCACCGCATCCCCAGCAT GCCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCAGA ATAGAATGACACCTACTCAGACAATGCGATGCAATTTCCTCATTTTATTAGGAAAGGACA GTGGGAGTGGCACCTTCCAGGGTCAAGGAAGGCACGGGGGAGGGGCAAACAACAGAT GGCTGGCAACTAGAAGGCACAGACGCGTGGGCCCCTGCAGTCGTACCGTGAGTAATAA TGCGATCGTCGTACCGTGAGTAATAATGCGCGGTTCGTACCGTGAGTAATAATGCGATC GTCGTACCGTGAGTAATAATGCGCTGCAGGTGTAGTGTTTCCTACTTTATGGAGTGATG TAGTGTTTCCTACTTTATGGACGGTTGTAGTGTTTCCTACTTTATGGAATCGTGTAGTGTT TCCTACTTTATGGAGGCTAGCCTAGGCGGGGAGGCGGCCCAAAGGGAGATCCGACTC GTCTGAGGGCGAAGGCGAAGACGCGGAAGAGGCCGCAGAGCCGGCAGCAGGCCGCG GGAAGGAAGGTCCGCTGGATTGAGGGCCGAAGGGACGTAGCAGAAGGACGTCCCGCG CAGAATCCAGGTGGCAACACAGGCGAGCAGCCATGGAAAGGACGTCAGCTTCCCCGAC AACACCACGGAATTGTCAGTGCCCAACAGCCGAGCCCCTGTCCAGCAGCGGGCAAGG CAGGCGGCGATGAGTTCCGCCGTGGCAATAGGGAGGGGGAAAGCGAAAGTCCCGGAA AGGAGCTGACAGGTGGTGGCAATGCCCCAACCAGTGGGGGTTGCGTCAGCAAACACA GTGCACACCACGCCACGTTGCCTGACAACGGGCCACAACTCCTCATAAAGAGACAGCA ACCAGGATTTATACAAGGAGGAGAAAATGAAAGCCATACGGGAAGCAATAGCATGATAC AAAGGCATTAAAGCAGCGTATCCACATAGCGTAAAAGGAGCAACATAGTTAAGAATACC AGTCAATCTTTCACAAATTTTGTAATCCAGAGGTTGATTGTCGACTCACAGGTTCTGTGT GCCGGCCTGCACAGACACCATGGAGAAGTAGATGCCCTTCTGGGCCAGCAGTTGCTG GTGGGTGCCGTGCTCTTTCACTCTGCCGTTCTGGAACACCACGATGAGGTCGGCATTC TGGA TGGTGCTCA GCCGGTGA GCGA TCA CGA TA CA GGTTCTTCCCTCGCGGGCCTTA T CCAGAGCCTCTTGCACCACCTTCTCGCTCTCGGTGTCCAGAGCAGAGGTGGCCTCATC CAGGA GCAGAA TCTGA GGCTGTCTGA TCAGGGCGCGAGCGA TAGCAA TCCTCTGTTT CTGGCCTCCGCTCAGCTGTGTGCCCTTGTCGCCCACTCTTGTCTCGTATTTGTGGGGC A GTGTCTCGA TGAA GGGGTGAATGTTGGCA GCCTTGGCA GCGGA CA CAA TCTCGTCC TGGGA CA CGA CTCTGCTGTTGTCGCCA TA GGCAA TA TTCTCGGCAA TGCTGCAGTCGA A CA GGA TGGGTTCTTGA GA CACGA TTCCCAGCTGA GCCCTCA GCCA CTGCA CGTTCA GCTTTTTGGCTTCTTGGCCATCCAGCAGGACGGTTCCAGCCAGAGGATCGTAGAACC GTTCCA GCAGCTGCA CCA CTGTA GA CTTGCCA CA TCCGCTA GA GCCCA CGA GA GCCA GA GTCTGGCCCTTCTTCACTTCCAGGCTCA GTCCCTGCA GAA CTGGCA CGTTGGCTCT GGTGGGGTAGTTGAACACCACCTCGTTAAAGGTGATGTTGCCCTCGAACTTATCTGGC TTCA GGCCTTCCTCGCTGTA GGA GTCGA TCA GGGGCTGTCTCTCGAA CA GCA TGAA CA GGTGTGCGGCAGACAGCTTGGCCTTGGCGTAGTCAGGGGCAAAGCTAGAGGCATGTC CCA GA GCCA CGGCTCCAAA CA CGA TGGCGGA GAA CA CCA GGA TCA CGTCCCGGAA T CTCATGTGGCCGTTCACGATCAGGTAGGCGCCGAATCTAAAGCAGCCGGCGTAGCTA AAGTA CA TGAAGGCCTGGCTGA TGCTGAA GGTGA TGCCGTAGA TGTGGGCCTTCTGC A CGCTGTTTCTGTA GGGGCCGTA CA GCTTTTCCACGTA CA TA GA CTCGAA CTTCCGCT CTTGGGTCAGAGA CA CCACGGTCCGGA TA TTCTCGA TGGCCTCGGTGGCGA TCTTGC CTGCGGCCTCA A GTTCTTTCTTGTCCCGCTTGGCGTTTCCGGCCA GCAA CTTCA TTTCC A CGA TGCCGGA CA CGGCAA TGA TA GGCA CCA CGGCCA GCA GGA GCA GTGTCA GCTG CCA TCCGTAGA TGAAGCTAATGA TGA TGCCGGTGCCGA GA TTGGCGA TA TTCTGGGC GA TCA GGGCCA GTCTTGTGCCTGTA GCGCCTTGCACCTGA GCA GCA TCTGTGGCCA G TCTGGTAGA CA GGGCGCCA GTGCTGTTCTTGTGGTCGTCGA A CCA A CTCA TGTCCTGT CTCA GCA TGGCTTTGAA GGCCA TGGA TCTCA GTCTCCGA GTCA GGA TCTCCCCGGCCT TGCCGA A GGTGA A GCCTTGCA GGA A GA A TGTA A A GA A GCTG A TAA TCCCCA GGA AC A GGAAGATCAGGCTAAAGATATTACACTTCTGCTGCTTCACGGCGTCGTCGCCAGGGC CAA AG A TGGCGA TAA TCTCGCTAAA GA TCA CGGA GA A GGCGGGCTGCA GTCCA CCA T TAGCAA TGGCA CA CACGGTGCCCA CGA CGA A GT A GGGCCA CTCGGTCTTGTTCAGTT TCA GCA CTTTCA GGAAGGA CA CA GGA GGCA CA TTGGCTTCCA GTCCGTCTGTCTCGA CGTCCA GGGA CTTCTGGCA CA TCTGGCTGTTCTTCA GA TTCTTCTGGGTGCTGTGTCTGAA CA GCCGGGA CTTCCA GCCA TTA GGGGCCA TCCTGGTGGCGGCCTTCTCGTCA TTC A GCTCGAATTCCTCGCTCTGGA TCTGA GA GCCGCTGGTCTGCA TGTTGA CCAGCTTGA A GT A CA CGCCTTCTTTCTTCA TCAGCTCGCTGTGGGA GCCCTGTTCCA CGA TCA CGCC A TCTTCGAATCCGGCGA TCA CA TCGGCA TTCCGCACGGTGGA CAGTCTGTGGGCAA T CA CGA TGGTGGTTCTGCCCTCTCTAGCCTTGTCCA GGGCA GCCTGCA CTTCTGCCTCA GACTCTGTATCCAGGGCGCTTGTGGCTTCGTCCAGCAGCAGGATCTTAGGGTTCCGCA CGA GTGCTCTGGCAA TGGCGA TTCTCTGCTTCTGTCCGCCA GA CA GTTGGGCGCCTCT TTCTCCCA CAA GGGTGTCGAA TTTCTGGGGCA GCTTCA TGA TGAA CTCGTA GGCGTTG GCCTCTTTCA CGGCTTTCTTGA TCTCGTCCA TGGTCA CGTTGCCTCTGCCGTA GCAGA T GTTCTCGGCGA TTGTGGTGCTGAA CA GCA CA GGCTCTTGGGA CA CCA CTCCGA TGA T CTCGCGCAGGTAGTTCACGTTGAAGTTCCGGATGTCCTGGCCGTCGA TGTTGA TTGTG CCCTCGTCAGGGTCGTA CA GTCTCTGGA TCA GCTGCA CGGTGGTGCTCTTGCCA CA G CCA GA AG A TCCCACGA GGGCCA CTGTCTGTCCGCTCTGCACCTTCA GGTTCA GGCCC TTC A GGA TCTTCA CGTTGGCCCGGCTA GGGTA GCTGAAGTGCACGTCGTTGAA TTCCA GGTTGCCCTTGA TGCTA TCAGGCTTGTGGCCCCGCTCGCTGAAGCTGTCGA TCTTGGG GTTGTTGTCGA TGA TA TCGAA GA TCA CGTA TGCGGCGCCTCTGGCA TTAGCAAAGGCG TCGA TA CAAGGA GCGGCCTGTCCGA CGCTAAA GGCGCCGA TCAGGA TGGA GA AG A A CACGGTCA TGGCGTTGCCGA TGGTGTACTCTTTGCTGA TGA CCA GGGTA GA GCCGTA CCAAAA GGCCA GGGCGTA GCTGGCGTA GA TCAGCAGGAAGGCGA TTCCCA TA GA GA TGTTGGCGCTGA TGGCCTTCTTGA TGCCAA TCTCTTTGGCGTTCTCGAGGTGCTTCTGG TA TCTTTCCAGTTCTTTGTTCTGGCCGCCAAA GGCGA TCA CGGTTCTGA TGGCTCCCA GGGCTTCTTCAGCAACGGCGCCAGCTTTAGCGTAAGCGGCCAGCTCTTTGTCGCTGA A GGCGCTCA GAA TCTTGGCCCA CA CA GCAGCA GA CA GTCCCAGGA TGGGA GA GA TG GCCA TGA TCACGA GGGTCA GCTTCCA GCCTCTGA TGAA GCCCACGA TAAAGCCGGCG AAGAAGGTGGCCACGGCCTGGAAGAACATTCCCACTTTGTCGCCGATGCCTTCGCTG A TCTTGCTGA TGTCGTCGGTCA GTCTGGTGTTCA GCTCGGTGGTGTCGTTGA TGTCGA ACCA TCCGA TCTCTTGCCGCAGGA TGGCGTGGAAGAACTTCTGCCGGA TCTTTCTGA T CTGCCGGCCA GCGGCCA GTGTCCAAAA GGA CACCTGGA TA TA GGCGGCCA CCA GAA CGCCAGCTCCAAGGCCGCTGTAGTAGTAGGCGTATCTGGTCATTTCCTCTTCCAGGAT CTTGCCGGGGTTCAGCAGGCTCAGGCTGAAATTCACGGGGAAGCTGAAGTTGCCGGC GGTGTCCACGAACTTGTCGGTCA TCTCGCCGAACACGA TCA TCATCAGGGGCAGTCC A GA TCCGTGGGCGA TA GCCA TGA TGGTGCCCA GGCTCA TGAA CA GCTTGTCCTGCCA GTCGCTGTA CCGGAACAGGGTCA GCA CGCCGA TCA TCTTCA CGGTCTTGGTCTTTTTC CGCTTCTGCTTGCTGCTGATGCCCAGCTCAAAATCGCCCTCGGCAGATGTAGGCCTCC A A GCGGTGCCA TTCTTGGCG GCTTCCA GA TCCA TGGTGGCGGATCACCGGTCTCAGC GGGAGGAGAAACCACAGCCTCAGAACCAAGTACACCCTCTCCGGCGGCCCGGCGCA CGAGTCGCGGGGCCCGGGGGCACTGGGTGGAGGTCCGGCCACTCCCGCCCCGCGCC CCACGTCCCGGGTCTCCCTTCGAGGCCAGACGCGCCCGGACCTCTCTCGGCTCGAAC CTCGCGCGTGTCTGGCAGGGCCTCTGGACGCGCGGGCGCTGCAGCAGAGGGGCCTG GACTTTGCTCGCCGCGGCCTCGCCCCCGCCCCCGCCCCCGCCCCCCACGCGCGTCC GGCCCGGGAAACGCCTACCGTTGCAGCCAGGGCGAGGGCCGCGGCCCGCGAGAGC AGCTCCCGCCGCCCGCCCAGCTCTCTCCCCAGGCTAGGCGCGGGCCAGGGGCGCAG CCTGCCGCTCTTGCCGCCGGGGCGCCCGGCTGGGGCCGCCGCCCACCCTGGCGACC CGCCAGAACCAGCCCGGTCGCAGGATAAGACCGAGATCAGGCTGGAGCCTCGGGTG CCGAGAAAGCCTCGCCTGATGGGCCACCCCCTCGAGGCCTCTCACACTACCTAAACC ACGCCAGGACAACCTCTGCTCCTCTCCACCGAAATTCCAAGGGGTCGAGTGGATGTT GGAGGTGGCA TGGGCCCA GA GA GGTCTCTGA CCTCTGCCCCA GCTCCAAGGTCA GCA GGCAGGGAGGGCTGTGTGTTTGCTGTTTGCTGCTTGCAATGTTTGCCCATTTTAGGGG ATATGCTTGAATTCGAGCTCGGTACCTTTAAGACCAATGACTTACAAGGCAGCTGTAGAT CTTAGCCACTTTTTAAAAGAAAAGGGGGGACTGGAAGGGCTAATTCACTCCCAACGAAG ACAAGATCTGCTTTTTGCTTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGG AGCTCTCTGGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTG CTTCAAGTAGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACC CTTTTAGTCAGTGTGGAAAATCTCTAGCAGTAGTAGTTCATGTCATCTTATTATTCAGTAT TTATAACTTGCAAAGAAATGAATATCAGAGAGTGAGAGGAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGC ATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGCTCTAGCTATC CCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGC CCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAG CTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGGACGTACCCAATTCGCCC TATAGTGAGTCGTATTACGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGA AAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGC GTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGG CGAATGGGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCG CAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTT CCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTA GGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGG TTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCA CGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCT ATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGAT TTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTTAGGTGGCACTT TTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTA TCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATAGCACCTAGATCAAGAGAC AGGATGAGGATCGTTTCGCATGATTGAACAAGATGGATTGCACGCAGGTTCTCCGGCC GCTTGGGTGGAGAGGCTATTCGGCTATGACTGGGCACAACAGACAATCGGCTGCTCTG ATGCCGCCGTGTTCCGGCTGTCAGCGCAGGGGCGCCCGGTTCTTTTTGTCAAGACCGA CCTGTCCGGTGCCCTGAATGAACTGCAAGACGAGGCAGCGCGGCTATCGTGGCTGGC CACGACGGGCGTTCCTTGCGCAGCTGTGCTCGACGTTGTCACTGAAGCGGGAAGGGA CTGGCTGCTATTGGGCGAAGTGCCGGGGCAGGATCTCCTGTCATCTCACCTTGCTCCT GCCGAGAAAGTATCCATCATGGCTGATGCAATGCGGCGGCTGCATACGCTTGATCCGG CTACCTGCCCATTCGACCACCAAGCGAAACATCGCATCGAGCGAGCACGTACTCGGAT GGAAGCCGGTCTTGTCGATCAGGATGATCTGGACGAAGAGCATCAGGGGCTCGCGCCA GCCGAACTGTTCGCCAGGCTCAAGGCGAGCATGCCCGACGGCGAGGATCTCGTCGTG ACCCATGGCGATGCCTGCTTGCCGAATATCATGGTGGAAAATGGCCGCTTTTCTGGATT CATCGACTGTGGCCGGCTGGGTGTGGCGGACCGCTATCAGGACATAGCGTTGGCTACC CGTGATATTGCTGAAGAGCTTGGCGGCGAATGGGCTGACCGCTTCCTCGTGCTTTACG GTATCGCCGCTCCCGATTCGCAGCGCATCGCCTTCTATCGCCTTCTTGACGAGTTCTTC TGAATTATTAACGCTTACAATTTCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGT ATTTCACACCGCATCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTT TATTTTTCTAAATACATTCAAATATGTATCCGCTCATGACCAAAATCCCTTAACGTGAGTT TTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTT TTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTT GTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCG CAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCT GTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGG CGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAG CGGTCGGGCTGAACGGGGGGGTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTAC ACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGA GAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGG GAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTG ACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCC AGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTT CCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATAC CGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGA GCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGG CACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTT AGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGT GGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAA GCGCGCAATTAACCCTCACTAAAGGGAACAAAAGCTGGAGCTGCAAGCTTGG

[0295] Annotations74..657: CMV promoter - allows expression of the vector genome

[0296] 658..5452: Vector genome

[0297] 672..852: 5' LTR (truncated)- truncated 5' long terminal repeat (LTR) from HIV-1 899..1024: HIV-1 packaging signal of human immunodeficiency virus type 1 1517..1750: RRE_ The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm

[0298] 2246..2363: cPPT / CTS_ central polypurine tract and central termination sequence of HIV- 1

[0299] 2394..2618: bGH poly(A) signal_ bovine growth hormone polyadenylation signal 2637..2736: 4x miR-126-3pT binding site (shown underlined)

[0300] 2744..2847: 4x miR142 binding site (shown underlined)

[0301] 2859..3447: WPRE_ woodchuck hepatitis virus posttranscriptional regulatory element 3454..7293: coABCB4_ codon optimised version of the human ABCB4 gene (shown bold and in italics)

[0302] 7311..7849: LPa_ novel promoter (shown in bold)

[0303] 7858..8048: HCR_ hApoE-HCR (Apolipoprotein E Hepatic Control Region) (shown bold and underlined)

[0304] 8141..8374: 3' LTR (AU3)_ self-inactivating 3' long terminal repeat (LTR) from HIV-1 8446..8567: SV40 poly(A) signal_ SV40 polyadenylation signal

[0305] 8568..74: Part of the plasmid needed for bacterial work

[0306] SEQ ID NO: 34 - miR-wtA4-LPaH, Version 3 backbone - wtABCB4 CCATTGCATACGTTGTATCCATATCATAATATGTACATTTATATTGGCTCATGTCCAACAT TACCGCCATGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATT AGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTG GCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTA ACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCA CTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACG GTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGG CAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATC AATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGT CAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTC CGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGA GCTCGTTTAGTGAACCGGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCT GGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGT AGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGT CAGTGTGGAAAATCTCTAGCAGTGGCGCCCGAACAGGGACTTGAAAGCGAAAGGGAAA CCAGAGGAGCTCTCTCGACGCAGGACTCGGCTTGCTGAAGCGCGCACGGCAAGAGGC GAGGGGCGGCGACTGGTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGAAGGAGA GAGATGGGTGCGAGAGCGTCAGTATTAAGCGGGGGAGAATTAGATCGCGATGGGAAAA AATTCGGTTAAGGCCAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTATGGGCAA GCAGGGAGCTAGAACGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGT AGACAAATACTGGGACAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATC ATTATATAATACAGTAGCAACCCTCTATTGTGTGCATCAAAGGATAGAGATAAAAGACAC CAAGGAAGCTTTAGACAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCACAG CAAGCGGCCGCTGATCTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAGT GAATTATATAAATATAAAGTAGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGCA AAGAGAAGAGTGGTGCAGAGAGAAAAAAGAGCAGTGGGAATAGGAGCTTTGTTCCTTG GGTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACA GGCCAGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTG AGGCGCAACAGCATCTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAG AATCCTGGCTGTGGAAAGATACCTAAAGGATCAACAGCTCCTGGGGATTTGGGGTTGCT CTGGAAAACTCATTTGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTCTGGAACAGATTTGGAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTAC ACAAGCTTAATACACTCCTTAATTGAAGAATCGCAAAACCAGCAAGAAAAGAATGAACAA GAATTATTGGAATTAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACATAACAAATTGG CTGTGGTATATAAAATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTT TTGCTGTACTTTCTATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGA CCCACCTCCCAACCCCGAGGGGACCCGACAGGCCCGAAGGAATAGAAGAAGAAGGTG GAGAGAGAGACAGAGACAGATCCATTCGATTAGTGAACGGATCTCGACGGTATCGGTTA ACTTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGACA TAATAGCAACAGACATACAAACTAAAGAATTACAAAAACAAATTACAAAAATTCAAAATTT TATCGATCACGAGACTAGCCTCGAGGATATCCCATAGAGCCCACCGCATCCCCAGCAT GCCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCAGA ATAGAATGACACCTACTCAGACAATGCGATGCAATTTCCTCATTTTATTAGGAAAGGACA GTGGGAGTGGCACCTTCCAGGGTCAAGGAAGGCACGGGGGAGGGGCAAACAACAGAT GGCTGGCAACTAGAAGGCACAGACGCGTGGGCCCCTGCAGTCGTACCGTGAGTAATAA TGCGATCGTCGTACCGTGAGTAATAATGCGCGGTTCGTACCGTGAGTAATAATGCGATC GTCGTACCGTGAGTAATAATGCGCTGCAGGTGTAGTGTTTCCTACTTTATGGAGTGATG TAGTGTTTCCTACTTTATGGACGGTTGTAGTGTTTCCTACTTTATGGAATCGTGTAGTGTT TCCTACTTTATGGAGGCTAGCCTAGGCGGGGAGGCGGCCCAAAGGGAGATCCGACTC GTCTGAGGGCGAAGGCGAAGACGCGGAAGAGGCCGCAGAGCCGGCAGCAGGCCGCG GGAAGGAAGGTCCGCTGGATTGAGGGCCGAAGGGACGTAGCAGAAGGACGTCCCGCG CAGAATCCAGGTGGCAACACAGGCGAGCAGCCATGGAAAGGACGTCAGCTTCCCCGAC AACACCACGGAATTGTCAGTGCCCAACAGCCGAGCCCCTGTCCAGCAGCGGGCAAGG CAGGCGGCGATGAGTTCCGCCGTGGCAATAGGGAGGGGGAAAGCGAAAGTCCCGGAA AGGAGCTGACAGGTGGTGGCAATGCCCCAACCAGTGGGGGTTGCGTCAGCAAACACA GTGCACACCACGCCACGTTGCCTGACAACGGGCCACAACTCCTCATAAAGAGACAGCA ACCAGGATTTATACAAGGAGGAGAAAATGAAAGCCATACGGGAAGCAATAGCATGATAC AAAGGCATTAAAGCAGCGTATCCACATAGCGTAAAAGGAGCAACATAGTTAAGAATACC AGTCAATCTTTCACAAATTTTGTAATCCAGAGGTTGATTGCTCATAAGTTCTGTGTCCCA GCCTGGA CA CTGA CCA TTGAAAAA TA GA TGCCTTTCTGTGCCA GCA GCTGCTGA TGCG TGCCA TGCTCCTTGA CTCTCCCA TTCTGAAA CACCA CT A TTAAGTCTGCA TTCTGGA TG GTGGA CA GGCGGTGA GCA A TCA CAA TGC A GGTGCGGCCTTCTCTGGCTTTGTCCA GG GCTTCTTGGA CAA CCTTTTCA CTTTCA GT A TCCA GA GCTGA TGTAGCTTCA TCCAA CA G GA GGA TTTGAGGTTGTCTGA TGA GGGCTCGGGCAA TA GCAA TCCTCTGTTTTTGA CCT CCTGA GA GCTGAGTCCCCTT A TCTCCCA CTCTTGTTTCA TA TTTGTGGGGTAACGTCTC GA TGAAA GGA TGTA TGTTGGCA GCTTTGGCTGCA CTCA CAA TTTCA TCCTGTGA TA CA A CCCGGCTGTTGTCTCCA TA GGCAA TA TTCTCGGCAA TGCTGCAGTCAAATAGGA TA G GCTCCTGA GA CACGA TTCCGAGTTGAGCTCTGA GCCA CTGGA CA TTGAGTTTCTTTGC TTCTTGA CCA TCGA GAA GCA CTGTCCCCGCCAA GGGGTCGTA GAACCGCTCCA GGA G CTGGA CCACCGTGCTCTTCCCA CA GCCACTGCTGCCCA CCA GGGCTAGTGTCTGGCC TTTCTTCA CCTCCA GGCTCA GCCCCTGAA GCA CTGGCA CGTTTGCTCGGGTGGGA TA G TTGAACACGA CTTCA TTAAATGTTA TA TTTCCTTCAAA TTTA TCA GGCTTCA GCCCCTCT TCA CTGTA GCTGTCAA TCA GA GGTTGTCTTTCAAA CA GCA TGA A TAA GTGGGCTGCAG A CA GCTTA GCTTTAGCA TAGTCTGGAGCAAATGAA CTGGCA TGTCCTA GA GCCA CTGC A CCAAA TA CAA TTGCA GAAAACACCA GAATAA CA TCTCTGAA GCGCA TA TGTCCA TTC AC A A TGA GA TA TGC ACC AAA TCGAAAACAA CCGGCA TA GGAAAAA TACATAAATGCT TGTGA GA TA CTAAAAGTAA TTCCA TAG A TGTGTGCCTTCTGCA CA GAATTCCTGTAA G GTCCA TACAA TTTTTCAA CA TA CA TTGA TTCAAATTTTCTTTCCTGGGTCAAA GA CA CA ACTGTCCTAATATTTTCTATTGCCTCTGTTGCAATCTTTCCAGCAGCTTCCAGTTCTTTT TTA TCTCTTTTGGCA TTTCCAGCCAACAA TTTCA TTTCA A CAA TTCCTGA CACAGCAAT A A TTGGAACAA CTGCTAA TA GCAA TA GGGTTAA CTGCCAA CCGTA GA TAAA TGA TA TG ATAATACCAGTTCCAAGGTTAGCTATATTCTGTGCAATTAAAGCCAACCTGGTTCCTGT GGCTCCTTGGA CTTGGGCAGCA TCTGTGGCAAGTCTTGTA GAAA GTGCA CCAGTACTG TTTTTATGGTCATCAAACCAGCTCATGTCCTGTCTTAGCATTGCTTTAAAAGCCATTGA CCGCAGTCTTCTGGTGAGGATCTCGCCAGCTTTCCCAAACGTGAAACCCTGAAGGAA GAAA GTAAAAAAA GAAA TAATTCCCA GAAA TAA GA AAA TCAAA GA GAA TA TGTTGCACTTCTGCTGCTTCA CTGCA TCA TCGCCTGGTCCAAAAA TCGCTA TGA TCTCTGA GA AT A TGA CTGAAAATGCCGGCTGAA GCCCCCCA TTGGCAA TGGCACA TACTGTTCCCA CGA CAAAGTA GGGCCA TTCTGTTTTA TTCAGTTTCAGGA CCTTCA GAAA GGA CA CTGGTGG CA CA TTTGCTTCAAGTCCA TCGGTTTCCACA TCAAGGCTCTTCTGA CACA TTTGTGAAT TTTTAA GGTTTTTCTGAGTA GAA TGCCTAAA TA GGCGA GA TTTCCA GCCA TTTGGGGCC A TTCTA GTGGCAGCCTTTTCA TCA TTTAGTTCAAA TTCTTCTGA CTGGA TCTGGCTTCCT GA TGTCTGCA TGTTGA CAAGTTTGAAGTA CA CCCCTTCCTTCTTCA TCAGTTCGCTGTG GCTTCCTTGCTCCACAA TTACTCCA TCCTCAAA CCCA GCGA TGA CA TCTGCATTTCGG A CCGTA GA CA GTCGGTGTGCTA TCA CAATGGTGGTCCGGCCTTCTCTGGCCTTA TCCA GA GCTGCCTGTA CCTCA GCTTCA CTTTCTGTGTCCAA TGCTGA CGTGGCCTCA TCCA G CA GAA GGA TCTTGGGGTTGCGAACCA GGGCA CGTGCAATGGCGA TCCTCTGCTTCTG CCCA CCA CTCA GCTGGGCCCCTCTCTCTCCAA CCA GGGTGTCAAA TTTCTGTGGTAA T TTCA TGA TAAA CTCA TA GGCGTTGGCCTCTTTGA CA GCTTTCTTTA TCTCA TCCA TGGTT ACATTTCCACGGCCATAACAAATATTTTCAGCAATTGTGGTGGAAAACAGCACCGGCT CCTGA CTCACCA CA CCAA TGA TTTCCCTCA GA TAGTTTA CA TTAAA GTTCCTAATA TCC TGCCCA TCAA TGTTAA TTGTGCCCTCA TCA GGGTCA TA GAGCCTCTGTA TCA GCTGGA CCGTTGTGCTCTTCCCA CA GCCA CT A CTTCCAA CCA GGGCCA CCGTCTGCCCACTCTG CA CCTTCA GGTTGA GGCCCTTCAA GA TCTTGA CGTTA GCTCGA GAA GGGTAAGAAAA GTGAACA TCA TTGAACTCCAAA TTCCCTTTGA TGCTGTCTGGTTTGTGTCCTCTCTCTG AAAAA CTGTCAA TTTTA GGA TTA TTA TCAA TAA TA TCA A A GA TCA CA TA TGCTGCTCCT CTTGCATTGGCAAAAGCATCAATACATGGGGCAGCCTGGCCAACACTGAAAGCTCCA A TTA GGA TTGAAAAAAAAACTGTCA TTGCA TTTCCAA TA GT A TA TTCTTTTGA TA TGA CT A GA GTGGA TCCA TA CCA GAA GGCCAGTGCA TA TGA TGC A TA TA TTA A CA GGA A GGCA A TA CCCA TGGAAA TGTTTGCTGAAA TA GCTTTTTTAA TTCCAA TCTCTTTGGCA TTTTCT AAA TGTTTCTGA TACCTTTCCA GCTCTTTGTTCTGGCCCCCGAAA GCTA TCA CAGTCCT GA TGGCCCCCA GA GCCTCTTCTGCCA CGGCGCCTGCTTTTGCA TAA GCA GCTAGTTCT TTGTCACTAAATGCCGAGAGTATCTTTGCCCAAACGGCTGCAGAGAGTCCTAGAATAG GGCTGA TGGCCA TTA TCA CAAGGGTGA GCTTCCA TCCTCTGA TGAATCCCA CT A TGA A TCCTGCAAAAAA CGTGGCTA CTGCTTGAAA GA AC A TTCCAA CCTTGTCA CCAA TTCCT TCA CTGA TTTTGGA GA TGTCA TCTGTTA GCCGCGTA TTGA GTTCAGTGGTGTCGTTGA T GTCAAA CCA TCCTA TTTCCTGTCGTA GAA TAGCA TGA AAAAA CTTCTGCCTAA TTTTCC TGATCTGCCGACCAGCTGCCAAAGTCCAAAATGAAACTTGTATATAGGCAGCAACAA GAA CTCCA GCACCCAA TCCTGAGTAGTAA TA TGC A TA TCTAGTCA TTTCTTCTTCCA GA ATTTTGCCTGGATTTAGCAGCGACAAGGAAAAGTTCACTGGAAAGGAGAAGTTTCCTG CAGTA TCAACAAATTTGTCAGTCA TCTCTCCAAA TA CT A TCA TCA TGA GGGGGA GA CC TGA TCCGTGAGCTA TGGCCA TGA TGGT ACCCAGCGA CA TAAA CAA TTTA TCCTGCCAA TCGGAGTA TCGAAA CAA TGTTAA TA CTCCA A TCA TTTTCA CTGTCTTCGTTTTTTTCCTT TTTTGTTTGCTGCTGA TGCCCA GTTCAAA GTCGCCCTCCGCGCTCGTGGGGCGCCA GG CTGTTCCGTTCTTTGCCGCCTCAAGATCCATGG GGCGGA CACCGG CTCAGCGGGA GGAGAAACCACAGCCTCAGAACCAAGTACACCCTCTCCGGCGGCCCGGCGCACGAG TCGCGGGGCCCGGGGGCACTGGGTGGAGGTCCGGCCACTCCCGCCCCGCGCCCCAC GTCCCGGGTCTCCCTTCGAGGCCAGACGCGCCCGGACCTCTCTCGGCTCGAACCTCG CGCGTGTCTGGCAGGGCCTCTGGACGCGCGGGCGCTGCAGCAGAGGGGCCTGGACT TTGCTCGCCGCGGCCTCGCCCCCGCCCCCGCCCCCGCCCCCCACGCGCGTCCGGCC CGGGAAACGCCTACCGTTGCAGCCAGGGCGAGGGCCGCGGCCCGCGAGAGCAGCTC CCGCCGCCCGCCCAGCTCTCTCCCCAGGCTAGGCGCGGGCCAGGGGCGCAGCCTGC CGCTCTTGCCGCCGGGGCGCCCGGCTGGGGCCGCCGCCCACCCTGGCGACCCGCCA GAACCAGCCCGGTCGCAGGATAAGACCGAGATCAGGCTGGAGCCTCGGGTGCCGAG AAAGCCTCGCCTGATGGGCCACCCCCTCGAGGCCTCTCACACTACCTAAACCACGCC AGGACAACCTCTGCTCCTCTCCACCGAAATTCCAAGGGGTCGAGTGGATGTTGGAGG TGGCATGGGCCCAGAGAGGTCTCTGACCTCTGCCCCAGCTCCAAGGTCAGCAGGCAG GGAGGGCTGTGTGTTTGCTGTTTGCTGCTTGCAATGTTTGCCCATTTTAGGGGATATGC TTGAATTCGAGCTCGGTACCTTTAAGACCAATGACTTACAAGGCAGCTGTAGATCTTAGC CACTTTTTAAAAGAAAAGGGGGGACTGGAAGGGCTAATTCACTCCCAACGAAGACAAGA TCTGCTTTTTGCTTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAA GTAGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTA GTCAGTGTGGAAAATCTCTAGCAGTAGTAGTTCATGTCATCTTATTATTCAGTATTTATAA CTTGCAAAGAAATGAATATCAGAGAGTGAGAGGAACTTGTTTATTGCAGCTTATAATGGT TACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTA GTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGCTCTAGCTATCCCGCC CCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCAT GGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTATT CCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGGACGTACCCAATTCGCCCTATA GTGAGTCGTATTACGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAAC CCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAA TAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAA TGGGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGC GTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTT TCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGG TTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCA CGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTT CTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTC TTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAA CAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTTAGGTGGCACTTTTCG GGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCC GCTCATGAGACAATAACCCTGATAAATGCTTCAATAATAGCACCTAGATCAAGAGACAG GATGAGGATCGTTTCGCATGATTGAACAAGATGGATTGCACGCAGGTTCTCCGGCCGCT TGGGTGGAGAGGCTATTCGGCTATGACTGGGCACAACAGACAATCGGCTGCTCTGATG CCGCCGTGTTCCGGCTGTCAGCGCAGGGGCGCCCGGTTCTTTTTGTCAAGACCGACCT GTCCGGTGCCCTGAATGAACTGCAAGACGAGGCAGCGCGGCTATCGTGGCTGGCCAC GACGGGCGTTCCTTGCGCAGCTGTGCTCGACGTTGTCACTGAAGCGGGAAGGGACTG GCTGCTATTGGGCGAAGTGCCGGGGCAGGATCTCCTGTCATCTCACCTTGCTCCTGCC GAGAAAGTATCCATCATGGCTGATGCAATGCGGCGGCTGCATACGCTTGATCCGGCTA CCTGCCCATTCGACCACCAAGCGAAACATCGCATCGAGCGAGCACGTACTCGGATGGA AGCCGGTCTTGTCGATCAGGATGATCTGGACGAAGAGCATCAGGGGCTCGCGCCAGCC GAACTGTTCGCCAGGCTCAAGGCGAGCATGCCCGACGGCGAGGATCTCGTCGTGACC CATGGCGATGCCTGCTTGCCGAATATCATGGTGGAAAATGGCCGCTTTTCTGGATTCAT CGACTGTGGCCGGCTGGGTGTGGCGGACCGCTATCAGGACATAGCGTTGGCTACCCG TGATATTGCTGAAGAGCTTGGCGGCGAATGGGCTGACCGCTTCCTCGTGCTTTACGGTA TCGCCGCTCCCGATTCGCAGCGCATCGCCTTCTATCGCCTTCTTGACGAGTTCTTCTGA ATTATTAACGCTTACAATTTCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTT CACACCGCATCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATT TTTCTAAATACATTCAAATATGTATCCGCTCATGACCAAAATCCCTTAACGTGAGTTTTCG TTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTT CTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTT GCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGA TACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAG CACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGAT AAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGT CGGGCTGAACGGGGGGGTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCG AACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAA GGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGC TTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTT GAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCA ACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTG CGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCT CGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCG CCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCAC GACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAA TTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCGC GCAATTAACCCTCACTAAAGGGAACAAAAGCTGGAGCTGCAAGCTTGGAAAATCAAAGA GAATATGTTGCACTTCTGCTGCTTCACTGCATCATCGCCTGGTCCAAAAATCGCTATGAT CTCTGAGAATATGACTGAAAATGCCGGCTGAAGCCCCCCATTGGCAATGGCACATACTG TTCCCACGACAAAGTAGGGCCATTCTGTTTTATTCAGTTTCAGGACCTTCAGAAA

[0307] Annotations

[0308] 74..657: CMV promoter - allows expression of the vector genome

[0309] 658..5452: Vector genome

[0310] 672..852: 5' LTR (truncated)- truncated 5' long terminal repeat (LTR) from HIV-1 899..1024: HIV-1 packaging signal of human immunodeficiency virus type 1 1517..1750: RRE_ The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm

[0311] 2246..2363: cPPT / CTS_ central polypurine tract and central termination sequence of HIV- 1

[0312] 2394..2618: bGH poly(A) signal_ bovine growth hormone polyadenylation signal 2637..2736: 4x miR-126-3pT binding site (shown underlined)

[0313] 2744..2847: 4x miR142 binding site (shown underlined)

[0314] 2859..3447: WPRE_ woodchuck hepatitis virus posttranscriptional regulatory element 3450..7289: wtABCB4_ wild type version of the human ABCB4 gene (shown in bold and italics)

[0315] 7307..7845: LPa promoter_ novel promoter (shown in bold)

[0316] 7854..8044: HCR_ hApoE-HCR (Apolipoprotein E Hepatic Control Region) (shown in bold and underlined)

[0317] 8137..8370: 3' LTR (AU3)_ self-inactivating 3' long terminal repeat (LTR) from HIV-1 8442..8563: SV40 poly(A) signal_ SV40 polyadenylation signal

[0318] 8564..74: Part of the plasmid needed for bacterial work

[0319] SEQ ID NO: 35 - miR-cA4-LPa CCATTGCATACGTTGTATCCATATCATAATATGTACATTTATATTGGCTCATGTCCAACAT TACCGCCATGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATT AGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTG GCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTA ACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCA CTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACG GTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGG CAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATC AATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGT CAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTC CGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGA GCTCGTTTAGTGAACCGGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCT GGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGT AGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGT CAGTGTGGAAAATCTCTAGCAGTGGCGCCCGAACAGGGACTTGAAAGCGAAAGGGAAA CCAGAGGAGCTCTCTCGACGCAGGACTCGGCTTGCTGAAGCGCGCACGGCAAGAGGC GAGGGGCGGCGACTGGTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGAAGGAGA GAGATGGGTGCGAGAGCGTCAGTATTAAGCGGGGGAGAATTAGATCGCGATGGGAAAA AATTCGGTTAAGGCCAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTATGGGCAA GCAGGGAGCTAGAACGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGT AGACAAATACTGGGACAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATC ATTATATAATACAGTAGCAACCCTCTATTGTGTGCATCAAAGGATAGAGATAAAAGACAC CAAGGAAGCTTTAGACAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCACAG CAAGCGGCCGCTGATCTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAGTGAATTATATAAATATAAAGTAGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGCA AAGAGAAGAGTGGTGCAGAGAGAAAAAAGAGCAGTGGGAATAGGAGCTTTGTTCCTTG GGTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACA GGCCAGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTG AGGCGCAACAGCATCTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAG AATCCTGGCTGTGGAAAGATACCTAAAGGATCAACAGCTCCTGGGGATTTGGGGTTGCT CTGGAAAACTCATTTGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTC TGGAACAGATTTGGAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTAC ACAAGCTTAATACACTCCTTAATTGAAGAATCGCAAAACCAGCAAGAAAAGAATGAACAA GAATTATTGGAATTAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACATAACAAATTGG CTGTGGTATATAAAATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTT TTGCTGTACTTTCTATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGA CCCACCTCCCAACCCCGAGGGGACCCGACAGGCCCGAAGGAATAGAAGAAGAAGGTG GAGAGAGAGACAGAGACAGATCCATTCGATTAGTGAACGGATCTCGACGGTATCGGTTA ACTTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGACA TAATAGCAACAGACATACAAACTAAAGAATTACAAAAACAAATTACAAAAATTCAAAATTT TATCGATCACGAGACTAGCCTCGAGGATATCCCATAGAGCCCACCGCATCCCCAGCAT GCCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCAGA ATAGAATGACACCTACTCAGACAATGCGATGCAATTTCCTCATTTTATTAGGAAAGGACA GTGGGAGTGGCACCTTCCAGGGTCAAGGAAGGCACGGGGGAGGGGCAAACAACAGAT GGCTGGCAACTAGAAGGCACAGACGCGTGGGCCCCTGCAGTCGTACCGTGAGTAATAA TGCGATCGTCGTACCGTGAGTAATAATGCGCGGTTCGTACCGTGAGTAATAATGCGATC GTCGTACCGTGAGTAATAATGCGCTGCAGGTGTAGTGTTTCCTACTTTATGGAGTGATG TAGTGTTTCCTACTTTATGGACGGTTGTAGTGTTTCCTACTTTATGGAATCGTGTAGTGTT TCCTACTTTATGGAGGCTAGCCTAGGCGGGGAGGCGGCCCAAAGGGAGATCCGACTC GTCTGAGGGCGAAGGCGAAGACGCGGAAGAGGCCGCAGAGCCGGCAGCAGGCCGCG GGAAGGAAGGTCCGCTGGATTGAGGGCCGAAGGGACGTAGCAGAAGGACGTCCCGCG CAGAATCCAGGTGGCAACACAGGCGAGCAGCCATGGAAAGGACGTCAGCTTCCCCGAC AACACCACGGAATTGTCAGTGCCCAACAGCCGAGCCCCTGTCCAGCAGCGGGCAAGG CAGGCGGCGATGAGTTCCGCCGTGGCAATAGGGAGGGGGAAAGCGAAAGTCCCGGAA AGGAGCTGACAGGTGGTGGCAATGCCCCAACCAGTGGGGGTTGCGTCAGCAAACACA GTGCACACCACGCCACGTTGCCTGACAACGGGCCACAACTCCTCATAAAGAGACAGCA ACCAGGATTTATACAAGGAGGAGAAAATGAAAGCCATACGGGAAGCAATAGCATGATAC AAAGGCATTAAAGCAGCGTATCCACATAGCGTAAAAGGAGCAACATAGTTAAGAATACC AGTCAATCTTTCACAAATTTTGTAATCCAGAGGTTGATTGTCGACTCACAGGTTCTGTGT GCCGGCCTGCACAGACACCATGGAGAAGTAGATGCCCTTCTGGGCCAGCAGTTGCTG GTGGGTGCCGTGCTCTTTCACTCTGCCGTTCTGGAACACCACGATGAGGTCGGCATTC TGGA TGGTGCTCA GCCGGTGA GCGA TCA CGA TA CA GGTTCTTCCCTCGCGGGCCTTA T CCAGAGCCTCTTGCACCACCTTCTCGCTCTCGGTGTCCAGAGCAGAGGTGGCCTCATC CA GGA GCAGAA TCTGA GGCTGTCTGA TCA GGGCGCGA GCGA TAGCAA TCCTCTGTTT CTGGCCTCCGCTCAGCTGTGTGCCCTTGTCGCCCACTCTTGTCTCGTATTTGTGGGGC A GTGTCTCGA TGAA GGGGTGA A TGTTGGCAGCCTTGGCA GCGGA CA CAA TCTCGTCC TGGGA CA CGA CTCTGCTGTTGTCGCCA TA GGCAA TA TTCTCGGCAA TGCTGCAGTCGA A CA GGA TGGGTTCTTGA GA CACGA TTCCCAGCTGA GCCCTCA GCCA CTGCA CGTTCA GCTTTTTGGCTTCTTGGCCA TCCA GCA GGA CGGTTCCA GCCA GA GGA TCGTA GA ACC GTTCCA GCAGCTGCA CCA CTGTA GA CTTGCCA CA TCCGCTA GA GCCCA CGA GA GCCA GAGTCTGGCCCTTCTTCACTTCCAGGCTCAGTCCCTGCAGAACTGGCACGTTGGCTCT GGTGGGGTAGTTGAACACCACCTCGTTAAAGGTGATGTTGCCCTCGAACTTATCTGGC TTCA GGCCTTCCTCGCTGTA GGA GTCGA TCA GGGGCTGTCTCTCGAA CA GCA TGAA CA GGTGTGCGGCAGACAGCTTGGCCTTGGCGTAGTCAGGGGCAAAGCTAGAGGCATGTC CCA GA GCCA CGGCTCCAAA CA CGA TGGCGGA GAA CA CCA GGA TCA CGTCCCGGAA T CTCATGTGGCCGTTCACGATCAGGTAGGCGCCGAATCTAAAGCAGCCGGCGTAGCTA AAGTA CA TGAAGGCCTGGCTGA TGCTGAA GGTGA TGCCGTAGA TGTGGGCCTTCTGC A CGCTGTTTCTGTA GGGGCCGTA CA GCTTTTCCACGTA CA TA GA CTCGAA CTTCCGCT CTTGGGTCAGAGA CA CCACGGTCCGGA TA TTCTCGA TGGCCTCGGTGGCGA TCTTGCCTGCGGCCTCAA GTTCTTTCTTGTCCCGCTTGGCGTTTCCGGCCA GCAA CTTCA TTTCC A CGA TGCCGGA CA CGGCAA TGA TA GGCA CCA CGGCCA GCA GGA GCA GTGTCA GCTG CCA TCCGTAGA TGAAGCTAATGA TGA TGCCGGTGCCGA GA TTGGCGA TA TTCTGGGC GA TCA GGGCCA GTCTTGTGCCTGTA GCGCCTTGCACCTGA GCA GCA TCTGTGGCCA G TCTGGTAGA CA GGGCGCCA GTGCTGTTCTTGTGGTCGTCGAA CCA A CTCA TGTCCTGT CTCA GCA TGGCTTTGAA GGCCA TGGA TCTCA GTCTCCGA GTCA GGA TCTCCCCGGCCT TGCCGAA GGTGAA GCCTTGCA GGA A GAA TGTAAA GAA GCTG A TAA TCCCCA GGA AC A GGAAGATCAGGCTAAAGATATTACACTTCTGCTGCTTCACGGCGTCGTCGCCAGGGC CAAAGA TGGCGA TAA TCTCGCTAAA GA TCA CGGA GAA GGCGGGCTGCA GTCCA CCA T TAGCAA TGGCA CA CACGGTGCCCA CGA CGA A GT A GGGCCA CTCGGTCTTGTTCAGTT TCA GCA CTTTCA GGAAGGA CA CA GGA GGCA CA TTGGCTTCCA GTCCGTCTGTCTCGA CGTCCA GGGA CTTCTGGCA CA TCTGGCTGTTCTTCA GA TTCTTCTGGGTGCTGTGTCTG A A CA GCCGGGA CTTCCA GCCA TTA GGGGCCA TCCTGGTGGCGGCCTTCTCGTCA TTC A GCTCGAA TTCCTCGCTCTGGA TCTGA GA GCCGCTGGTCTGCA TGTTGA CCAGCTTGA A GT A CA CGCCTTCTTTCTTCA TCAGCTCGCTGTGGGA GCCCTGTTCCA CGA TCA CGCC A TCTTCGAATCCGGCGA TCA CA TCGGCA TTCCGCACGGTGGA CAGTCTGTGGGCAA T CA CGA TGGTGGTTCTGCCCTCTCTA GCCTTGTCCA GGGCA GCCTGCA CTTCTGCCTCA GACTCTGTATCCAGGGCGCTTGTGGCTTCGTCCAGCAGCAGGATCTTAGGGTTCCGCA CGA GTGCTCTGGCAA TGGCGA TTCTCTGCTTCTGTCCGCCA GA CA GTTGGGCGCCTCT TTCTCCCA CAA GGGTGTCGAA TTTCTGGGGCA GCTTCA TGA TGA A CTCGTA GGCGTTG GCCTCTTTCA CGGCTTTCTTGA TCTCGTCCA TGGTCA CGTTGCCTCTGCCGTA GCAGA T GTTCTCGGCGA TTGTGGTGCTGAA CA GCA CA GGCTCTTGGGA CA CCA CTCCGA TGA T CTCGCGCAGGTAGTTCACGTTGAAGTTCCGGATGTCCTGGCCGTCGA TGTTGA TTGTG CCCTCGTCAGGGTCGTA CA GTCTCTGGA TCA GCTGCA CGGTGGTGCTCTTGCCA CA G CCA GA AG A TCCCACGA GGGCCA CTGTCTGTCCGCTCTGCACCTTCA GGTTCA GGCCC TTC A GGA TCTTCA CGTTGGCCCGGCTA GGGTA GCTGAAGTGCACGTCGTTGAA TTCCA GGTTGCCCTTGA TGCTA TCAGGCTTGTGGCCCCGCTCGCTGAAGCTGTCGA TCTTGGG GTTGTTGTCGA TGA TA TCGAA GA TCA CGTA TGCGGCGCCTCTGGCA TTAGCAAAGGCG TCGA TA CAAGGA GCGGCCTGTCCGA CGCTAAA GGCGCCGA TCAGGA TGGA GA AG A A CACGGTCA TGGCGTTGCCGA TGGTGTACTCTTTGCTGA TGACCAGGGTAGAGCCGTA CCAAAA GGCCA GGGCGTA GCTGGCGTA GA TCAGCAGGAAGGCGA TTCCCA TA GA GA TGTTGGCGCTGA TGGCCTTCTTGA TGCCAATCTCTTTGGCGTTCTCGAGGTGCTTCTGG TA TCTTTCCAGTTCTTTGTTCTGGCCGCCAAA GGCGA TCA CGGTTCTGA TGGCTCCCA GGGCTTCTTCA GCAA CGGCGCCA GCTTTAGCGTAA GCGGCCA GCTCTTTGTCGCTGA A GGCGCTCA GAA TCTTGGCCCA CA CA GCAGCA GA CA GTCCCAGGA TGGGA GA GA TG GCCA TGA TCACGA GGGTCA GCTTCCA GCCTCTGA TGAA GCCCACGA TAAAGCCGGCG AAGAAGGTGGCCACGGCCTGGAAGAACA TTCCCA CTTTGTCGCCGATGCCTTCGCTG A TCTTGCTGA TGTCGTCGGTCA GTCTGGTGTTCA GCTCGGTGGTGTCGTTGA TGTCGA ACCA TCCGA TCTCTTGCCGCAGGA TGGCGTGGAAGAACTTCTGCCGGA TCTTTCTGA T CTGCCGGCCA GCGGCCA GTGTCCAAAA GGA CA CCTGGA TA TA GG CGGCCA CCA GAA CGCCAGCTCCAAGGCCGCTGTAGTAGTAGGCGTATCTGGTCATTTCCTCTTCCAGGAT CTTGCCGGGGTTCAGCAGGCTCAGGCTGAAATTCACGGGGAAGCTGAAGTTGCCGGC GGTGTCCACGAACTTGTCGGTCA TCTCGCCGAACACGA TCA TCATCAGGGGCAGTCC A GA TCCGTGGGCGA TA GCCA TGA TGGTGCCCA GGCTCA TGAA CA GCTTGTCCTGCCA GTCGCTGTA CCGGAACAGGGTCA GCA CGCCGA TCA TCTTCA CGGTCTTGGTCTTTTTC CGCTTCTGCTTGCTGCTGATGCCCAGCTCAAAATCGCCCTCGGCAGATGTAGGCCTCC A A GCGGTGCCA TTCTTGGCG GCTTCCA GA TCCA TGGTGGCGGA TCA CCGG TCTCA GC GGGAGGA GA A A CCA CA GCCTCA GAA CCAAGTA CACCCTCTCCGGCGGCCCGGCGCA CGAGTCGCGGGGCCCGGGGGCACTGGGTGGAGGTCCGGCCACTCCCGCCCCGCGCC CCA CGTCCCGGGTCTCCCTTCGA GGCCA GACGCGCCCGGA CCTCTCTCGGCTCGAA C CTCGCGCGTGTCTGGCAGGGCCTCTGGACGCGCGGGCGCTGCAGCAGAGGGGCCTG GACTTTGCTCGCCGCGGCCTCGCCCCCGCCCCCGCCCCCGCCCCCCACGCGCGTCC GGCCCGGGAAA CGCCTA CCGTTGCA GCCA GGGCGA GGGCCGCGGCCCGCGA GAGC A GCTCCCGCCGCCCGCCCA GCTCTCTCCCCA GGCTA GGCGCGGGCCA GGGGCGCAG CCTGCCGCTCTTGCCGCCGGGGCGCCCGGCTGGGGCCGCCGCCCACCCTGGCGACCCGCCA G CCA GCCCGGTCGCA GGA TA AG A CCGA GA TCA GGCTGGA GCCTCGGGTG CCGAGAAA GCCTCGCCTGA TGGGCCA CCCCCTCGAGGGGA TA TGCTTGAA TTCGAGC TCGGTACCTTTAAGACCAATGACTTACAAGGCAGCTGTAGATCTTAGCCACTTTTTAAAA GAAAAGGGGGGACTGGAAGGGCTAATTCACTCCCAACGAAGACAAGATCTGCTTTTTGC TTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCTGGCTAACTAG GGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCC CGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAA AATCTCTAGCAGTAGTAGTTCATGTCATCTTATTATTCAGTATTTATAACTTGCAAAGAAA TGAATATCAGAGAGTGAGAGGAACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGC AA TAGCA TCA CAAA TTTCA CAAA TAAA GCA TTTTTTTCA C TGCA TTCTA G TTG TGGTTTG T CCAAACTCA TCAA TGTA TCTTATCA TGTCTGGCTCTAGCTATCCCGCCCCTAACTCCGCC CATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATGGCTGACTAATTT TTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTATTCCAGAAGTAGTGA GGAGGCTTTTTTGGAGGCCTAGGGA CG TACCCAA TTCGCCC TA TAG TGA G TCG TA TTA C GCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCC AACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCC CGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAATGGGACGCGCCCT GTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACAC TTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTC GCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGC TTTACGGCACCTCGA CCCCAAAAAA C TTG A TTA GGGTGA TGG TTCA CG TAGTGGGCCA T CGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGA CTCTTGTTCCAAACTGGAACAACACTCAACCCTA TCTCGGTCTA TTCTTTTGA TTTA TAAG GGA TTTTGCCGA TTTCGGCCTA TTGGTTAAAAAA TGAGCTGA TTTAACAAAAA TTTAACG CGAA TTTTAA CAAAA TA TTAA CGCTTACAA TTTAGG TGGCAC TTTTCGGGGAAA TG TGCG CGGAACCCCTA TTTG TTTA TTTTTCTAAA TACA TTCAAA TA TG TA TCCGCTCA TGAGA CAA TAACCCTGATAAATGCTTCAATAATAGCACCTAGATCAAGAGACAGGATGAGGATCGTTT CGCATGATTGAACAAGATGGATTGCACGCAGGTTCTCCGGCCGCTTGGGTGGAGAGGC TATTCGGCTATGACTGGGCACAACAGACAATCGGCTGCTCTGATGCCGCCGTGTTCCG GCTGTCAGCGCAGGGGCGCCCGGTTCTTTTTGTCAAGACCGACCTGTCCGGTGCCCTG AATGAACTGCAAGACGAGGCAGCGCGGCTATCGTGGCTGGCCACGACGGGCGTTCCTT GCGCAGCTGTGCTCGACGTTGTCACTGAAGCGGGAAGGGACTGGCTGCTATTGGGCG AAGTGCCGGGGCAGGATCTCCTGTCATCTCACCTTGCTCCTGCCGAGAAAGTATCCATC ATGGCTGATGCAATGCGGCGGCTGCATACGCTTGATCCGGCTACCTGCCCATTCGACC ACCAAGCGAAACATCGCATCGAGCGAGCACGTACTCGGATGGAAGCCGGTCTTGTCGA TCAGGATGATCTGGACGAAGAGCATCAGGGGCTCGCGCCAGCCGAACTGTTCGCCAG GCTCAAGGCGAGCATGCCCGACGGCGAGGATCTCGTCGTGACCCATGGCGATGCCTG CTTGCCGAA TA TCA TGGTGGAAAATGGCCGCTTTTCTGGA TTCATCGACTGTGGCCGGC TGGGTG TGGCGGA CCGCTA TCA GGA CA TAGCG TTGGCTA CCCG TGA TA TTGCTGAA GA GCTTGGCGGCGAATGGGCTGACCGCTTCCTCGTGCTTTACGGTATCGCCGCTCCCGAT TCGCAGCGCA TCGCCTTCTA TCGCCTTCTTGA CGAGTTC TTC TGAA TTA TTAACGC TTA C AA TTTCCTGA TGCGGTA TTTTCTCCTTACGCA TCTGTGCGGTATTTCACACCGCATCAGG TGGCACTTTTCGGGGAAA TG TGCG CGGAACCCCTA TTTGTTTA TTTTTCTAAA TACA TTC AAA TA TGTA TCCGCTCA TGACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTC AGACCCCGTAGAAAAGA TCAAAGGA TCTTCTTGAGA TCCTTTTTTTCTGCGCGTAATCTG CTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGC TACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTC TTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATAC CTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTAC CGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGG GGGGTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTA CAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTAT CCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAA CGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTT GTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGAT TCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAAC GACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACC GCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGAC TGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCAC CCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAA CAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCGCGCAATTAACCCTCA CTAAAGGGAACAAAAGCTGGAGCTGCAAGCTTGG

[0320] Annotations:

[0321] • 74..657: CMV promoter - allows expression of the vector genome

[0322] • 658..5452: Vector genome

[0323] • 672..852: 5' LTR (truncated)- truncated 5' long terminal repeat (LTR) from HIV-1

[0324] • 899..1024: HIV-1 Ψ_ packaging signal of human immunodeficiency virus type 1

[0325] • 1517..1750: RRE_ The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm

[0326] • 2246..2363: cPPT / CTS_ central polypurine tract and central termination sequence of HIV- 1

[0327] • 2394..2618: bGH poly(A) signal_ bovine growth hormone polyadenylation signal

[0328] • 2637..2736: 4x miR-126-3pT binding site (shown underlined)

[0329] • 2744..2847: 4x miR142 binding site (shown underlined)

[0330] • 2859..3447: WPRE_ woodchuck hepatitis virus posttranscriptional regulatory element • 3454..7293: coABCB4_ codon optimised version of the human ABCB4 gene (shown in bold and italics)

[0331] • 7311..7849: LPa_v3 promoter_ novel promoter derived from ABCB4 promoter (shown in bold)

[0332] • 7951..8184: 3' LTR (AU3)_ self-inactivating 3' long terminal repeat (LTR) from HIV-1 • 8256..8377: SV40 poly(A) signal_ SV40 polyadenylation signal

[0333] • 8378..74: Part of the plasmid needed for bacterial work

[0334] SEQ ID NO: 36 - miR-wA4-LPa CCATTGCATACGTTGTATCCATATCATAATATGTACATTTATATTGGCTCATGTCCAACAT TACCGCCATGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATT AGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTG GCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTA ACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCA CTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACG GTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGG CAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATC AATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGT CAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTC CGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGA GCTCGTTTAGTGAACCGGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCT GGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGT AGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGT CAGTGTGGAAAATCTCTAGCAGTGGCGCCCGAACAGGGACTTGAAAGCGAAAGGGAAA CCAGAGGAGCTCTCTCGACGCAGGACTCGGCTTGCTGAAGCGCGCACGGCAAGAGGC GAGGGGCGGCGACTGGTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGAAGGAGA GAGATGGGTGCGAGAGCGTCAGTATTAAGCGGGGGAGAATTAGATCGCGATGGGAAAA AATTCGGTTAAGGCCAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTATGGGCAA GCAGGGAGCTAGAACGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGT AGACAAATACTGGGACAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATC ATTATATAATACAGTAGCAACCCTCTATTGTGTGCATCAAAGGATAGAGATAAAAGACAC CAAGGAAGCTTTAGACAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCACAGCAAGCGGCCGCTGATCTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAGT GAATTATATAAATATAAAGTAGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGCA AAGAGAAGAGTGGTGCAGAGAGAAAAAAGAGCAGTGGGAATAGGAGCTTTGTTCCTTG GGTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACA GGCCAGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTG AGGCGCAACAGCATCTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAG AATCCTGGCTGTGGAAAGATACCTAAAGGATCAACAGCTCCTGGGGATTTGGGGTTGCT CTGGAAAACTCATTTGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTC TGGAACAGATTTGGAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTAC ACAAGCTTAATACACTCCTTAATTGAAGAATCGCAAAACCAGCAAGAAAAGAATGAACAA GAATTATTGGAATTAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACATAACAAATTGG CTGTGGTATATAAAATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTT TTGCTGTACTTTCTATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGA CCCACCTCCCAACCCCGAGGGGACCCGACAGGCCCGAAGGAATAGAAGAAGAAGGTG GAGAGAGAGACAGAGACAGATCCATTCGATTAGTGAACGGATCTCGACGGTATCGGTTA ACTTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGACA TAATAGCAACAGACATACAAACTAAAGAATTACAAAAACAAATTACAAAAATTCAAAATTT TATCGATCACGAGACTAGCCTCGAGGATATCCCATAGAGCCCACCGCATCCCCAGCAT GCCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCAGA ATAGAATGACACCTACTCAGACAATGCGATGCAATTTCCTCATTTTATTAGGAAAGGACA GTGGGAGTGGCACCTTCCAGGGTCAAGGAAGGCACGGGGGAGGGGCAAACAACAGAT GGCTGGCAACTAGAAGGCACAGACGCGTGGGCCCCTGCAGTCGTACCGTGAGTAATAA TGCGATCGTCGTACCGTGAGTAATAATGCGCGGTTCGTACCGTGAGTAATAATGCGATC GTCGTACCGTGAGTAATAATGCGCTGCAGGTGTAGTGTTTCCTACTTTATGGAGTGATG TAGTGTTTCCTACTTTATGGACGGTTGTAGTGTTTCCTACTTTATGGAATCGTGTAGTGTT TCCTACTTTATGGAGGCTAGCCTAGGCGGGGAGGCGGCCCAAAGGGAGATCCGACTC GTCTGAGGGCGAAGGCGAAGACGCGGAAGAGGCCGCAGAGCCGGCAGCAGGCCGCG GGAAGGAAGGTCCGCTGGATTGAGGGCCGAAGGGACGTAGCAGAAGGACGTCCCGCG CAGAATCCAGGTGGCAACACAGGCGAGCAGCCATGGAAAGGACGTCAGCTTCCCCGAC AACACCACGGAATTGTCAGTGCCCAACAGCCGAGCCCCTGTCCAGCAGCGGGCAAGG CAGGCGGCGATGAGTTCCGCCGTGGCAATAGGGAGGGGGAAAGCGAAAGTCCCGGAA AGGAGCTGACAGGTGGTGGCAATGCCCCAACCAGTGGGGGTTGCGTCAGCAAACACA GTGCACACCACGCCACGTTGCCTGACAACGGGCCACAACTCCTCATAAAGAGACAGCA ACCAGGATTTATACAAGGAGGAGAAAATGAAAGCCATACGGGAAGCAATAGCATGATAC AAAGGCATTAAAGCAGCGTATCCACATAGCGTAAAAGGAGCAACATAGTTAAGAATACC AGTCAATCTTTCACAAATTTTGTAATCCAGAGGTTGATTGTCGACCGACTTACTTGTACAGCTCGTCCATGCCGAGAGTGATCCCGGCGGCGGTCACGAACTCCAGCAGGACCATGT CCAGCCTGGACACTGACCATTGAAAAATAGATGCCTTTCTGTGCCAGCAGCTGCTGATG CGTGCCATGCTCCTTGACTCTCCCA TTCTGAAACACCACTA TTAAGTCTGCA TTCTGGA T GGTGGACAGGCGGTGAGCAATCACAATGCAGGTGCGGCCTTCTCTGGCTTTGTCCAGG GCTTCTTGGACAACCTTTTCACTTTCAGTA TCCAGAGCTGA TGTAGCTTCA TCCAACAGG AGGA TTTGAGGTTGTCTGATGAGGGCTCGGGCAA TAGCAA TCCTCTGTTTTTGACCTCC TGAGAGCTGAGTCCCCTTATCTCCCACTCTTGTTTCATATTTGTGGGGTAACGTCTCGAT GAAAGGA TGTA TGTTGGCAGCTTTGGCTGCACTCACAA TTTCA TCCTGTGA TACAACCC GGCTGTTGTCTCCATAGGCAA TATTCTCGGCAA TGCTGCAGTCAAA TAGGA TAGGCTCC TGAGACACGATTCCGAGTTGAGCTCTGAGCCACTGGACATTGAGTTTCTTTGCTTCTTG ACCATCGAGAAGCACTGTCCCCGCCAAGGGGTCGTAGAACCGCTCCAGGAGCTGGAC CACCGTGCTCTTCCCACAGCCACTGCTGCCCACCAGGGCTAGTGTCTGGCCTTTCTTCA CCTCCAGGCTCAGCCCCTGAAGCACTGGCACGTTTGCTCGGGTGGGATAGTTGAACAC GACTTCA TTAAA TGTTA TATTTCCTTCAAA TTTA TCAGGCTTCAGCCCCTCTTCACTGTAG CTGTCAATCAGAGGTTGTCTTTCAAACAGCATGAATAAGTGGGCTGCAGACAGCTTAGC TTTAGCATAGTCTGGAGCAAATGAACTGGCATGTCCTAGAGCCACTGCACCAAATACAA TTGCA GA AAA CA CCA GAA TA A CA TCTCTGAA GCGCATATG TCCA TTCA CAA TGA GA TA TG CACCAAA TCGAAAACAACCGGCATAGGAAAAA TACATAAA TGCTTGTGAGA TACTAAAAG TAA TTCCA TA GA TG TG TGCCTTCTGCA CA GAA TTCCTGTAA GG TCCA TA CAA TTTTTCAA CA TA CA TTGA TTCAAA TTTTCTTTCC TGGGTCAAA GA CACAACTGTCCTAA TA TTTTCTA TTGCCTCTGTTGCAATCTTTCCAGCAGCTTCCAGTTCTTTTTTATCTCTTTTGGCATTTCCA GCCAACAA TTTCA TTTCAACAATTCCTGACACAGCAA TAA TTGGAACAACTGCTAATAGC AA TAGGGTTAACTGCCAACCGTAGA TAAATGATA TGA TAA TACCAGTTCCAAGGTTAGCT ATATTCTGTGCAATTAAAGCCAACCTGGTTCCTGTGGCTCCTTGGACTTGGGCAGCATC TGTGGCAAGTCTTGTAGAAAGTGCACCAGTACTGTTTTTATGGTCATCAAACCAGCTCAT GTCCTGTCTTAGCATTGCTTTAAAAGCCATTGACCGCAGTCTTCTGGTGAGGATCTCGC CAGCTTTCCCAAACGTGAAACCCTGAAGGAAGAAAGTAAAAAAAGAAA TAA TTCCCAGA AATAAGAAAATCAAAGAGAATATGTTGCACTTCTGCTGCTTCACTGCATCATCGCCTGGT CCAAAAA TCGCTA TGA TCTCTGAGAA TA TGACTGAAAA TGCCGGCTGAAGCCCCCCA TT GGCAATGGCACATACTGTTCCCACGACAAAGTAGGGCCATTCTGTTTTATTCAGTTTCAG GACCTTCAGAAAGGACACTGGTGGCACATTTGCTTCAAGTCCATCGGTTTCCACATCAA GGCTCTTCTGACACATTTGTGAATTTTTAAGGTTTTTCTGAGTAGAATGCCTAAATAGGC GAGA TTTCCAGCCA TTTGGGGCCA TTCTAGTGGCAGCCTTTTCA TCA TTTAGTTCAAA TT CTTCTGACTGGA TCTGGCTTCCTGA TGTCTGCA TGTTGACAAGTTTGAAGTACACCCCTT CCTTCTTCATCAGTTCGCTGTGGCTTCCTTGCTCCACAATTACTCCATCCTCAAACCCAG CGATGACATCTGCATTTCGGACCGTAGACAGTCGGTGTGCTATCACAATGGTGGTCCG GCCTTCTCTGGCCTTATCCAGAGCTGCCTGTACCTCAGCTTCACTTTCTGTGTCCAATG CTGACGTGGCCTCATCCAGCAGAAGGATCTTGGGGTTGCGAACCAGGGCACGTGCAAT GGCGATCCTCTGCTTCTGCCCACCACTCAGCTGGGCCCCTCTCTCTCCAACCAGGGTG TCAAA TTTCTGTGGTAA TTTCA TGA TAAACTCA TAGGCGTTGGCCTCTTTGACAGCTTTCT TTA TCTCA TCCA TGGTTACA TTTCCACGGCCA TAA CAAA TA TTTTCAGCAA TTG TGG TGG AAAACAGCACCGGCTCCTGACTCACCACACCAA TGA TTTCCCTCAGA TAGTTTACATTAA AGTTCCTAATA TCCTGCCCA TCAATGTTAATTGTGCCCTCA TCAGGGTCA TAGAGCCTCT GTATCAGCTGGACCGTTGTGCTCTTCCCACAGCCACTACTTCCAACCAGGGCCACCGTC TGCCCACTCTGCACCTTCAGGTTGAGGCCCTTCAAGATCTTGACGTTAGCTCGAGAAGG GTAAGAAAAGTGAACATCATTGAACTCCAAATTCCCTTTGATGCTGTCTGGTTTGTGTCC TCTCTCTGAAAAACTGTCAA TTTTAGGA TTA TTA TCAA TAA TA TCAAAGA TCACA TA TGCT GCTCCTCTTGCATTGGCAAAAGCATCAATACATGGGGCAGCCTGGCCAACACTGAAAGC TCCAA TTAGGA TTGAAAAAAAAACTGTCA TTGCA TTTCCAA TAGTA TA TTCTTTTGA TA TG ACTAGAGTGGA TCCA TACCAGAAGGCCAGTGCA TATGATGCATA TA TTAACAGGAAGGC AA TACCCA TGG AAA TGTTTGCTGAAA TAGCTTTTTTAA TTCCAA TCTCTTTGGCA TTTTCT AAATGTTTCTGATACCTTTCCAGCTCTTTGTTCTGGCCCCCGAAAGCTATCACAGTCCTG ATGGCCCCCAGAGCCTCTTCTGCCACGGCGCCTGCTTTTGCATAAGCAGCTAGTTCTTT GTCACTAAATGCCGAGAGTATCTTTGCCCAAACGGCTGCAGAGAGTCCTAGAATAGGGC TGA TGGCCA TTA TCACAAGGGTGAGCTTCCATCCTCTGA TGAA TCCCACTA TGAA TCCT GCAAAAAACGTGGCTACTGCTTGAAAGAACATTCCAACCTTGTCACCAATTCCTTCACTG A TTTTG GA GA TG TCA TCTGTTAGCCGCGTA TTG A G TTCA GTGGTGTCG TTG A TG TCAAA C CATCCTATTTCCTGTCGTAGAATAGCATGAAAAAACTTCTGCCTAATTTTCCTGATCTGC CGACCAGCTGCCAAAGTCCAAAATGAAACTTGTATATAGGCAGCAACAAGAACTCCAGC ACCCAA TCCTGAGTA G TAA TA TGCA TA TC TA G TCA TTTCTTCTTCCAGAA TTTTGCCTGGA TTTAGCAGCGACAAGGAAAAGTTCACTGGAAAGGAGAAGTTTCCTGCAGTATCAACAAA TTTGTCAGTCA TCTCTCCAAATACTATCA TCA TGAGGGGGAGACCTGATCCGTGAGCTA T GGCCA TGA TGG TACCCA GCGA CA TAAA CAA TTTA TCCTGCCAA TCGGAGTA TCGAAACA A TG TTAA TACTCCAA TCA TTTTCA C TG TC TTCG TTTTTTTCC TTTTTTG TTTGCTGCTGA TG CCCAGTTCAAAGTCGCCCTCCGCGCTCGTGGGGCGCCAGGCTGTTCCGTTCTTTGCCG CCTCAAGATCCATGGTGGCGGATCACCGGTCTCAGCGGGAGGAGAAACCACAGCCTC AGAACCAAGTACACCCTCTCCGGCGGCCCGGCGCACGAGTCGCGGGGCCCGGGGGC ACTGGGTGGAGGTCCGGCCACTCCCGCCCCGCGCCCCACGTCCCGGGTCTCCCTTCG AGGCCAGACGCGCCCGGACCTCTCTCGGCTCGAACCTCGCGCGTGTCTGGCAGGGC CTCTGGACGCGCGGGCGCTGCAGCAGAGGGGCCTGGACTTTGCTCGCCGCGGCCTC GCCCCCGCCCCCGCCCCCGCCCCCCACGCGCGTCCGGCCCGGGAAACGCCTACCGT TGCAGCCAGGGCGAGGGCCGCGGCCCGCGAGAGCAGCTCCCGCCGCCCGCCCAGC TCTCTCCCCAGGCTAGGCGCGGGCCAGGGGCGCAGCCTGCCGCTCTTGCCGCCGGG GCGCCCGGCTGGGGCCGCCGCCCACCCTGGCGACCCGCCAGAACCAGCCCGGTCGC AGGATAAGACCGAGATCAGGCTGGAGCCTCGGGTGCCGAGAAAGCCTCGCCTGATGGGCCACCCCCTCGAGGCGGATATGCTTGAATTCGAGCTCGGTACCTTTAAGACCAATG ACTTACAAGGCAGCTGTAGATCTTAGCCACTTTTTAAAAGAAAAGGGGGGACTGGAAGG GCTAATTCACTCCCAACGAAGACAAGATCTGCTTTTTGCTTGTACTGGGTCTCTCTGGTT AGACCAGATCTGAGCCTGGGAGCTCTCTGGCTAACTAGGGAACCCACTGCTTAAGCCT CAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCCCGTCTGTTGTGTGACTCTGG TAACTAGAGATCCCTCAGACCCTTTTAGTCAGTGTGGAAAATCTCTAGCAGTAGTAGTTC ATGTCATCTTATTATTCAGTATTTATAACTTGCAAAGAAATGAATATCAGAGAGTGAGAGG AACTTGTTTATTGCAGCTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAA ATAAAGCATTTTTTTCACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTA TCATGTCTGGCTCTAGCTATCCCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCC CAGTTCCGCCCATTCTCCGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGA GGCCGCCTCGGCCTCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTA GGGACGTACCCAATTCGCCCTATAGTGAGTCGTATTACGCGCGCTCACTGGCCGTCGT TTTACAACGTCGTGACTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCAC ATCCCCCTTTCGCCAGCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCA ACAGTTGCGCAGCCTGAATGGCGAATGGGACGCGCCCTGTAGCGGCGCATTAAGCGC GGCGGGTGTGGTGGTTACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCC CGCTCCTTTCGCTTTCTTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAG CTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCC AAAAAACTTGATTAGGGTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTT TCGCCCTTTGACGTTGGAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAAC AACACTCAACCCTATCTCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGC CTATTGGTTAAAAAATGAGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTA ACGCTTACAATTTAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTAT TTTTCTAAATACATTCAAATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCA ATAATAGCACCTAGATCAAGAGACAGGATGAGGATCGTTTCGCATGATTGAACAAGATG GATTGCACGCAGGTTCTCCGGCCGCTTGGGTGGAGAGGCTATTCGGCTATGACTGGGC ACAACAGACAATCGGCTGCTCTGATGCCGCCGTGTTCCGGCTGTCAGCGCAGGGGCG CCCGGTTCTTTTTGTCAAGACCGACCTGTCCGGTGCCCTGAATGAACTGCAAGACGAG GCAGCGCGGCTATCGTGGCTGGCCACGACGGGCGTTCCTTGCGCAGCTGTGCTCGAC GTTGTCACTGAAGCGGGAAGGGACTGGCTGCTATTGGGCGAAGTGCCGGGGCAGGAT CTCCTGTCATCTCACCTTGCTCCTGCCGAGAAAGTATCCATCATGGCTGATGCAATGCG GCGGCTGCATACGCTTGATCCGGCTACCTGCCCATTCGACCACCAAGCGAAACATCGC ATCGAGCGAGCACGTACTCGGATGGAAGCCGGTCTTGTCGATCAGGATGATCTGGACG AAGAGCATCAGGGGCTCGCGCCAGCCGAACTGTTCGCCAGGCTCAAGGCGAGCATGC CCGACGGCGAGGATCTCGTCGTGACCCATGGCGATGCCTGCTTGCCGAATATCATGGT GGAAAATGGCCGCTTTTCTGGATTCATCGACTGTGGCCGGCTGGGTGTGGCGGACCGC TATCAGGACATAGCGTTGGCTACCCGTGATATTGCTGAAGAGCTTGGCGGCGAATGGG CTGACCGCTTCCTCGTGCTTTACGGTATCGCCGCTCCCGATTCGCAGCGCATCGCCTTC TATCGCCTTCTTGACGAGTTCTTCTGAATTATTAACGCTTACAATTTCCTGATGCGGTATT TTCTCCTTACGCATCTGTGCGGTATTTCACACCGCATCAGGTGGCACTTTTCGGGGAAA TGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATG ACCAAAATCCCTTAACGTGAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGAT CAAAGGATCTTCTTGAGATCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAA ACCACCGCTACCAGCGGTGGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGA AGGTAACTGGCTTCAGCAGAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAG TTAGGCCACCACTTCAAGAACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCT GTTACCAGTGGCTGCTGCCAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGA CGATAGTTACCGGATAAGGCGCAGCGGTCGGGCTGAACGGGGGGGTCGTGCACACAG CCCAGCTTGGAGCGAACGACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAG AAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGG TCGGAACAGGAGAGCGCACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAG TCCTGTCGGGTTTCGCCACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGG GGCGGAGCCTATGGAAAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTG CTGGCCTTTTGCTCACATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGA GTCAGTGAGCGAGGAAGCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCG TTGGCCGATTCATTAATGCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGT GAGCGCAACGCAATTAATGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTT TATGCTTCCGGCTCGTATGTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAA ACAGCTATGACCATGATTACGCCAAGCGCGCAATTAACCCTCACTAAAGGGAACAAAAG CTGGAGCTGCAAGCTTGG

[0335] Annotations:

[0336] • 74..657: CMV promoter - allows expression of the vector genome

[0337] • 658..5452: Vector genome

[0338] • 672..852: 5' LTR (truncated)- truncated 5' long terminal repeat (LTR) from HIV-1

[0339] • 899..1024: HIV-1 Ψ_ packaging signal of human immunodeficiency virus type 1

[0340] • 1517..1750: RRE_ The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm

[0341] • 2246..2363: cPPT / CTS_ central polypurine tract and central termination sequence of HIV- 1

[0342] • 2394..2618: bGH poly(A) signal_ bovine growth hormone polyadenylation signal

[0343] • 2637..2736: 4x miR-126-3pT binding site (shown underlined)

[0344] • 2744..2847: 4x miR142 binding site

[0345] • 2859..3447: WPRE_ woodchuck hepatitis virus posttranscriptional regulatory element • 3454..7293: wtABCB4_ wild type version of the human ABCB4 gene (shown in bold and italics)

[0346] • 7311..7849: LPa v3 promoter novel promoter derived from ABCB4 promoter (shown bold)

[0347] • 7952..8185: 3' LTR (AU3)_ self-inactivating 3' long terminal repeat (LTR) from HIV-1 • 8257..8378: SV40 poly(A) signal_ SV40 polyadenylation signal

[0348] • 8379..74: Part of the plasmid needed for bacterial work

[0349] SEQ ID NO: 39 - miR-GFP. Luc-LPaH CCATTGCATACGTTGTATCCATATCATAATATGTACATTTATATTGGCTCATGTCCAACAT TACCGCCATGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATT AGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTG GCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTA ACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCA CTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACG GTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGG CAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATC AATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGT CAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTC CGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGA GCTCGTTTAGTGAACCGGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCT GGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGT AGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGT CAGTGTGGAAAATCTCTAGCAGTGGCGCCCGAACAGGGACTTGAAAGCGAAAGGGAAA CCAGAGGAGCTCTCTCGACGCAGGACTCGGCTTGCTGAAGCGCGCACGGCAAGAGGC GAGGGGCGGCGACTGGTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGAAGGAGA GAGATGGGTGCGAGAGCGTCAGTATTAAGCGGGGGAGAATTAGATCGCGATGGGAAAA AATTCGGTTAAGGCCAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTATGGGCAA GCAGGGAGCTAGAACGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGT AGACAAATACTGGGACAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATC ATTATATAATACAGTAGCAACCCTCTATTGTGTGCATCAAAGGATAGAGATAAAAGACAC CAAGGAAGCTTTAGACAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCACAG CAAGCGGCCGCTGATCTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAGTGAATTATATAAATATAAAGTAGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGCA AAGAGAAGAGTGGTGCAGAGAGAAAAAAGAGCAGTGGGAATAGGAGCTTTGTTCCTTG GGTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACA GGCCAGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTG AGGCGCAACAGCATCTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAG AATCCTGGCTGTGGAAAGATACCTAAAGGATCAACAGCTCCTGGGGATTTGGGGTTGCT CTGGAAAACTCATTTGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTC TGGAACAGATTTGGAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTAC ACAAGCTTAATACACTCCTTAATTGAAGAATCGCAAAACCAGCAAGAAAAGAATGAACAA GAATTATTGGAATTAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACATAACAAATTGG CTGTGGTATATAAAATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTT TTGCTGTACTTTCTATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGA CCCACCTCCCAACCCCGAGGGGACCCGACAGGCCCGAAGGAATAGAAGAAGAAGGTG GAGAGAGAGACAGAGACAGATCCATTCGATTAGTGAACGGATCTCGACGGTATCGGTTA ACTTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGACA TAATAGCAACAGACATACAAACTAAAGAATTACAAAAACAAATTACAAAAATTCAAAATTT TATCGATCACGAGACTAGCCTCGAGGATATCCCATAGAGCCCACCGCATCCCCAGCAT GCCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCAGA ATAGAATGACACCTACTCAGACAATGCGATGCAATTTCCTCATTTTATTAGGAAAGGACA GTGGGAGTGGCACCTTCCAGGGTCAAGGAAGGCACGGGGGAGGGGCAAACAACAGAT GGCTGGCAACTAGAAGGCACAGACGCGTGGGCCCCTGCAGTCGTACCGTGAGTAATAA TGCGATCGTCGTACCGTGAGTAATAATGCGCGGTTCGTACCGTGAGTAATAATGCGATC GTCGTACCGTGAGTAATAATGCGCTGCAGGTGTAGTGTTTCCTACTTTATGGAGTGATG TAGTGTTTCCTACTTTATGGACGGTTGTAGTGTTTCCTACTTTATGGAATCGTGTAGTGTT TCCTACTTTATGGAGGCTAGCCTAGGCGGGGAGGCGGCCCAAAGGGAGATCCGACTC GTCTGAGGGCGAAGGCGAAGACGCGGAAGAGGCCGCAGAGCCGGCAGCAGGCCGCG GGAAGGAAGGTCCGCTGGATTGAGGGCCGAAGGGACGTAGCAGAAGGACGTCCCGCG CAGAATCCAGGTGGCAACACAGGCGAGCAGCCATGGAAAGGACGTCAGCTTCCCCGAC AACACCACGGAATTGTCAGTGCCCAACAGCCGAGCCCCTGTCCAGCAGCGGGCAAGG CAGGCGGCGATGAGTTCCGCCGTGGCAATAGGGAGGGGGAAAGCGAAAGTCCCGGAA AGGAGCTGACAGGTGGTGGCAATGCCCCAACCAGTGGGGGTTGCGTCAGCAAACACA GTGCACACCACGCCACGTTGCCTGACAACGGGCCACAACTCCTCATAAAGAGACAGCA ACCAGGATTTATACAAGGAGGAGAAAATGAAAGCCATACGGGAAGCAATAGCATGATAC AAAGGCATTAAAGCAGCGTATCCACATAGCGTAAAAGGAGCAACATAGTTAAGAATACC AGTCAATCTTTCACAAATTTTGTAATCCAG AGGTTG ATTGTCG ACCG AC TTA CTTGTA CA GCTCGTCCA TGCCGA GAGTGA TCCCGGCGGCGGTCA CGAA CTCCA GCA GGA CCA TGT GATCGCGCTTCTCGTTGGGGTCTTTGCTCAGGGCGGACTGGGTGCTCAGGTAGTGGTT GTCGGGCAGCAGCACGGGGCCGTCGCCGATGGGGGTGTTCTGCTGGTAGTGGTCGG CGA GCTGCACGCTGCCGTCCTCGA TGTTGTGGCGGA TCTTGAA GTTCA CCTTGA TGCC GTTCTTCTGCTTGTCGGCCA TGA TA TA GA CGTTGTGGCTGTTGTA GTTGTA CTCCA GCT TGTGCCCCA GGA TGTTGCCGTCCTCCTTGA A GTCGA TGCCCTTCA GCTCGA TGCGGTT CACCAGGGTGTCGCCCTCGAACTTCACCTCGGCGCGGGTCTTGTAGTTGCCGTCGTC CTTGAAGAAGATGGTGCGCTCCTGGACGTAGCCTTCGGGCATGGCGGACTTGAAGAA GTCGTGCTGCTTCATGTGGTCGGGGTAGCGGCTGAAGCACTGCACGCCGTAGGTCAG GGTGGTCA CGA GGGTGGGCCA GGGCACGGGCA GCTTGCCGGTGGTGCA GA TGA A CT TCAGGGTCAGCTTGCCGTAGGTGGCATCGCCCTCGCCCTCGCCGGACACGCTGAACT TGTGGCCGTTTACGTCGCCGTCCAGCTCGACCAGGATGGGCACCACCCCGGTGAACA GCTCCTCGCCCTTGCTCA CCA TGGGGCCGGGA TTTTCCTCGA CA TCTCCA CA GGTCA G CAA TGA GCCCCGGCCCTCTGCGCGGA TCTTGCCGCCCTTCTTGGCCTTGA TCA GGA TC TCCCGGATCTTTCTGGCGTCCAGCTTGCCGGTCAGGCCCTTGGGCACCTCGTCCACGA ACACCACGCCGCCTCTCAGCTTCTTGGCGGTGGTCACCTGGCTGGCCACGTAGTCCA CGA TCTCCTTCTCGGTCA TGGTCTTGCCGTGCTCCA GCA CCA CCACGGCGGCTGGCA GCTCTCCGGCGTCGTCGTCGGGCAGTCCGGCCACTCCGGCGTCCCGGATGTTGGGGT GCTGCAGCAGGATGCTCTCCAGCTCGGCTGGGGCCACCTGGTAGCCCTTGTACTTGA TCA GGCTCTTCA GCCGGTCCACGA TGA A GAA GTGCTCGTCCTCGTCCCAGTA GGCGATGTCGCCGCTGTGCAGCCAGCCGTCCTTGTCGATCAGGGCGTTGGTGGCCTCGGGGT TGTTCACGTA GCCGCTCA TGA TCA TGGGGCCTCTCA CGCA CA GCTCGCCTCTCTGGTT CA CGCCCA GGGTCTTGCCGGTGTCCA GGTCCACCA CCTTGGCCTCGAA GAAGGGCAC CACCTTGCCCACGGCTCCGGGCTTGTCGTCGCCCTCGGGGGTGATCAGGATGGCGCT GGTGGTCTCGGTCAGGCCGTAGCCCTGCCGGATGCCTGGCAGGTGGAACCGCTTGGC CACGGCCTCGCCCACCTCCTTGCTCAGTGGGGCTCCGCCAGAGGCGATCTCGTGCAG GTTGCTCAGGTCGTACTTGTCGATCAGGGTGCTCTTGGCGAAGAAGCTGAACAGGGT GGGCA CCA GCA GGGCGCTCTGGA TCTTGTAGTCCTGCA GGCTCCGCA GGAACAGCTC CTCCTCGAACCTGTACATCAGCACCACCCGGAAGCCGCAGATCAGGTAGCCCAGGGT GGTGAA CA TGCCGAAGCCGTGGTGGAA GGGCA CCA CGCTCA GGA TGGCGGTGTCGG GCTTGATCTGGTTGCCGAAGATGGGGTCTCTGGCGTGGCTGAACCGCACGCAGGCGG TCCGGTGGGGCAGGGCCACGCCCTTGGGCAGGCCGGTGCTGCCGCTGCTGTTCATGA TCAGGGCGATGGTCTTGTCCCGGTCGAAGCTCTCGGGCTTGAAGTCGTACTCGTTGAA GCCTGGGGGCAGGTGGCTGGTCACGAAGGTGTACATGCTCTGGAAGCCCTGGTAATC GGTCTTGCTGTCCA TGA TGA TGA TCTTCTGGA TGA TGGGCAGCTTCTTCTGCA CA TTCA GGA TCTTCTGCAGGCCCTTCTTGCTCA CGAACACCA CGGTGGGCTGGCTGA TGTTCA T GCTGTTCA GCAGCTCCCGCTCGTTGTAGA TGTCGTTGGCTGGGGCCACGGCCA CGCC GATGAACAGGGCTCCCAGCACGGGCATGAAGAACTGCAGGCTGTTCTCGCTGCACAC CACGATCCGGTGGTTGGTGTTCAGGCCGTACCGCTTCATGGCCTCGGCCAGCCGCAC GCTCATCTCGAAGTACTCGGCGTAGGTGATGTTCACCTCGATGTGGGCGTCGGTGAA GGCGA TGGTGCCTGGCA CCTGGGCGTACCGCTTCA TGGCCTTGTGCA GCTGCTCGCC GGCTGTGCCGTCCTCCA GGGGGTA TCTGGGGGCTGGTCCCTTCTTGA TGTTCTTGGCG TCCTCCA TGCTGCCGCCGCCGCTCTTGTCA TCGTCA TCCTTGTAATCGA TGTCA TGA TC TTTA TAA TCACCGTCA TGGTCTTTGTAGTCCATGGTGGCACCGGTCTCAGCGGGAGGA GAAACCACAGCCTCAGAACCAAGTACACCCTCTCCGGCGGCCCGGCGCACGAGTCG CGGGGCCCGGGGGCACTGGGTGGAGGTCCGGCCACTCCCGCCCCGCGCCCCACGTC CCGGGTCTCCCTTCGAGGCCAGACGCGCCCGGACCTCTCTCGGCTCGAACCTCGCGC GTGTCTGGCAGGGCCTCTGGACGCGCGGGCGCTGCAGCAGAGGGGCCTGGACTTTG CTCGCCGCGGCCTCGCCCCCGCCCCCGCCCCCGCCCCCCACGCGCGTCCGGCCCGG GAAACGCCTACCGTTGCAGCCAGGGCGAGGGCCGCGGCCCGCGAGAGCAGCTCCCG CCGCCCGCCCAGCTCTCTCCCCAGGCTAGGCGCGGGCCAGGGGCGCAGCCTGCCGC TCTTGCCGCCGGGGCGCCCGGCTGGGGCCGCCGCCCACCCTGGCGACCCGCCAGAA CCAGCCCGGTCGCAGGATAAGACCGAGATCAGGCTGGAGCCTCGGGTGCCGAGAAA GCCTCGCCTGATGGGCCACCCCCTCGAGGCCTCTCACACTACCTAAACCACGCCAGG ACAACCTCTGCTCCTCTCCACCGAAATTCCAAGGGGTCGAGTGGATGTTGGAGGTGG CATGGGCCCAGAGAGGTCTCTGACCTCTGCCCCAGCTCCAAGGTCAGCAGGCAGGG AGGGCTGTGTGTTTGCTGTTTGCTGCTTGCAATGTTTGCCCATTTTAGGGGATATGCTT GAATTCGAGCTCGGTACCTTTAAGACCAATGACTTACAAGGCAGCTGTAGATCTTAGCC ACTTTTTAAAAGAAAAGGGGGGACTGGAAGGGCTAATTCACTCCCAACGAAGACAAGAT CTGCTTTTTGCTTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTC TGGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAG TAGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAG TCAGTGTGGAAAATCTCTAGCAGTAGTAGTTCATGTCATCTTATTATTCAGTATTTATAAC TTGCAAAGAAATGAATATCAGAGAGTGAGAGGAACTTGTTTATTGCAGCTTATAATGGTT ACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACTGCATTCTAG TTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGCTCTAGCTATCCCGCCC CTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTCCGCCCCATG GCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCTGAGCTATTC CAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGGACGTACCCAATTCGCCCTATAGT GAGTCGTATTACGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTGGGAAAACC CTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCTGGCGTAAT AGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAATGGCGAAT GGGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTACGCGCAGCG TGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTCCCTTCCTTT CTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTGATGGTTCA CGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAGTCCACGTT CTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCGGTCTATTC TTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGCTGATTTAA CAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTTAGGTGGCACTTTTCG GGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATATGTATCC GCTCATGAGACAATAACCCTGATAAATGCTTCAATAATAGCACCTAGATCAAGAGACAG GATGAGGATCGTTTCGCATGATTGAACAAGATGGATTGCACGCAGGTTCTCCGGCCGCT TGGGTGGAGAGGCTATTCGGCTATGACTGGGCACAACAGACAATCGGCTGCTCTGATG CCGCCGTGTTCCGGCTGTCAGCGCAGGGGCGCCCGGTTCTTTTTGTCAAGACCGACCT GTCCGGTGCCCTGAATGAACTGCAAGACGAGGCAGCGCGGCTATCGTGGCTGGCCAC GACGGGCGTTCCTTGCGCAGCTGTGCTCGACGTTGTCACTGAAGCGGGAAGGGACTG GCTGCTATTGGGCGAAGTGCCGGGGCAGGATCTCCTGTCATCTCACCTTGCTCCTGCC GAGAAAGTATCCATCATGGCTGATGCAATGCGGCGGCTGCATACGCTTGATCCGGCTA CCTGCCCATTCGACCACCAAGCGAAACATCGCATCGAGCGAGCACGTACTCGGATGGA AGCCGGTCTTGTCGATCAGGATGATCTGGACGAAGAGCATCAGGGGCTCGCGCCAGCC GAACTGTTCGCCAGGCTCAAGGCGAGCATGCCCGACGGCGAGGATCTCGTCGTGACC CATGGCGATGCCTGCTTGCCGAATATCATGGTGGAAAATGGCCGCTTTTCTGGATTCAT CGACTGTGGCCGGCTGGGTGTGGCGGACCGCTATCAGGACATAGCGTTGGCTACCCG TGATATTGCTGAAGAGCTTGGCGGCGAATGGGCTGACCGCTTCCTCGTGCTTTACGGTA TCGCCGCTCCCGATTCGCAGCGCATCGCCTTCTATCGCCTTCTTGACGAGTTCTTCTGA ATTATTAACGCTTACAATTTCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCGGTATTT CACACCGCATCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATT TTTCTAAATACATTCAAATATGTATCCGCTCATGACCAAAATCCCTTAACGTGAGTTTTCG TTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCTTTTTTT CTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGTGGTTTGTTT GCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCAGAGCGCAGA TACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAGAACTCTGTAG CACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGCCAGTGGCGAT AAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAGGCGCAGCGGT CGGGCTGAACGGGGGGGTCGTGCACACAGCCCAGCTTGGAGCGAACGACCTACACCG AACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAA GGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCGCACGAGGGAGC TTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGCCACCTCTGACTT GAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGAAAAACGCCAGCA ACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCACATGTTCTTTCCTG CGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGAGCTGATACCGCT CGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAAGCGGAAGAGCG CCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAATGCAGCTGGCAC GACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAATGTGAGTTAGC TCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTATGTTGTGTGGAA TTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATTACGCCAAGCGC GCAATTAACCCTCACTAAAGGGAACAAAAGCTGGAGCTGCAAGCTTGG

[0350] Annotations:

[0351] • 74..657: CMV promoter - allows expression of the vector genome

[0352] • 658..7041: Vector genome

[0353] • 672..852: 5' LTR (truncated)- truncated 5' long terminal repeat (LTR) from HIV-1

[0354] • 899..1024: HIV-1 Ψ_ packaging signal of human immunodeficiency virus type 1

[0355] • 1517..1750: RRE_ The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm

[0356] • 2246..2363: cPPT / CTS_ central polypurine tract and central termination sequence of HIV- 1

[0357] • 2394..2618: bGH poly(A) signal_ bovine growth hormone polyadenylation signal

[0358] • 2637..2736: 4x miR-126-3pT binding site (shown underlined)• 2744..2847: 4x miR142 binding site

[0359] • 2859..3447: WPRE_ woodchuck hepatitis virus posttranscriptional regulatory element • 3458..5962: Luciferase. GFP (shown in bold and italics)

[0360] • 5978..6516: LPa_v3 promoter_ novel promoter derived from ABCB4 promoter (shown bold)

[0361] • 6525..6715: HCR_ hApoE-HCR (Apolipoprotein E Hepatic Control Region) (shown in bold and underlined)

[0362] • 6808..7041: 3' LTR (AU3)_ self-inactivating 3' long terminal repeat (LTR) from HIV-1 • 7113..7234: SV40 poly(A) signal_ SV40 polyadenylation signal

[0363] • 7235..74: Part of the plasmid needed for bacterial work

[0364] SEQ ID NO: 65 - Predicted ABCB4 natural promoter sequence and flanking region (1048bp)

[0365] TAGGAGCAGAGACAATTCAATACAGACAGAAGTCTTAGATGAGAGCAGTGAGCCAGGG CACTGGACTGGGACTCAGGAGGCTTCCCCTAGACTCTGGTTCCACCGATGCAGCCTCA GGCAGGACTTCACCTCTCTGGGCATCCGTTTCTTCATATGTTAAACATACGGGGTTTTAA TTAGATGATCGCTGAAGACCCCTCTAGCCCTAAAACTCTGTGTCTCTTAAGTGCTAAGAG GGCACCAACAGCGTTCCTCCTCCCCAAGGAGCATAATGTGATGGTTCCTGCCGGCCCT GGCTGACTCTCGCCGTCCTTGGAGATAATTGGGTTCAGTGCCACCTGGACCAGAACTG GGGATGCGGAAGCAAGAGGCGAGTCTATTGCTCTCTCTCGGTCCTGGGCCGCCCTGTG ATTGTTGGGCGTCCGGAAACTGTCTCCCCTATGGGTTTAAAAACAAAACTGAGCGCCCA TGGGGTGTGACAGTCATCTGCAGGGGCTTGGGTGGCCCATCAGGCGAGGCTTTCTCG GCACCCGAGGCTCCAGCCTGATCTCGGTCTTATCCTGCGACCGGGCTGGTTCTGGCGG GTCGCCAGGGTGGGCGGCGGCCCCAGCCGGGCGCCCCGGCGGCAAGAGCGGCAGG CTGCGCCCCTGGCCCGCGCCTAGCCTGGGGAGAGAGCTGGGCGGGCGGCGGGAGCT GCTCTCGCGGGCCGCGGCCCTCGCCCTGGCTGCAACGGTAGGCGTTTCCCGGGCCGG ACGCGCGTGGGGGGCGGGGGCGGGGGCGGGGGCGAGGCCGCGGCGAGCAAAGTCC AGGCCCCTCTGCTGCAGCGCCCGCGCGTCCAGAGGCCCTGCCAGACACGCGCGAGGT TCGAGGTGAGAGAGGTCCGGGCGCGTCTGGCCTCGAAGGGAGACCCGGGACGTGGG GCGCGGGGCGGGAGTGGCCGGACCTCCACCCAGTGCCCCCGGGCCCCGCGACTCGT GCGCCGGGCCGCCGGAGAGGGTGTACTTGGTTCTGAGGCTGTGGTTTCTCCTCAGGCT GAGATGGATCTTGA

[0366] SEQ ID NO: 66 - Region for enhancing expression TCCCCTAGACTCTGGTTCCACCGATGCAGCCTCAGGCAGGACTTCACCTCTCTGGGCAT CCGTTTCTTCATATGTTAAACATACGGGGTTTTAATTAGATGATCGCTGAAGACCCCTCT AGCCCTAAAACTCTGTGTCTCTTAAGTGCTAAGAGGGCACCAACAGCGTTCCTCCTCCC CAAGGAGCATAATGTGATGGTTCCTGCCGGCCCTGGCTGACTCTCGCCGTCCTTGGAG ATAATTGGGTTCAGTGCCACCTGGACCAGAACTGGGGATGCGGAAGCAAGA

[0367] SEQ ID NO: 67 - Reverse complement of region for enhancing expressionTCTTGCTTCCGCATCCCCAGTTCTGGTCCAGGTGGCACTGAACCCAATTATCTCCAAGG ACGGCGAGAGTCAGCCAGGGCCGGCAGGAACCATCACATTATGCTCCTTGGGGAGGA GGAACGCTGTTGGTGCCCTCTTAGCACTTAAGAGACACAGAGTTTTAGGGCTAGAGGG GTCTTCAGCGATCATCTAATTAAAACCCCGTATGTTTAACATATGAAGAAACGGATGCCC AGAGAGGTGAAGTCCTGCCTGAGGCTGCATCGGTGGAACCAGAGTCTAGGGGA

[0368] SEQ ID NO: 68 - Sequence corresponding to the natural human ABCB4 promoter sequence and flanking region (1048bp) with splice sites mutated, i.e., the donor splice site is not GT, and the acceptor splice site is not AG.

[0369] TAGGAGCAGAGACAATTCAATACAGACAGAAGTCTTAGATGAGAGCAGTGAGCCAGGG CACTGGACTGGGACTCAGGAGGCTTCCCCTAGACTCTGGTTCCACCGATGCAGCCTCA GGCAGGACTTCACCTCTCTGGGCATCCGTTTCTTCATATGTTAAACATACGGGGTTTTAA TTAGATGATCGCTGAAGACCCCTCTAGCCCTAAAACTCTGTGTCTCTTAAGTGCTAAGAG GGCACCAACAGCGTTCCTCCTCCCCAAGGAGCATAATGTGATGGTTCCTGCCGGCCCT GGCTGACTCTCGCCGTCCTTGGAGATAATTGGGTTCAGTGCCACCTGGACCAGAACTG GGGATGCGGAAGCAAGAGGCGAGTCTATTGCTCTCTCTCGGTCCTGGGCCGCCCTGTG ATTGTTGGGCGTCCGGAAACTGTCTCCCCTATGGGTTTAAAAACAAAACTGAGCGCCCA TGGGGTGTGACAGTCATCTGCAGGGGCTTGGGTGGCCCATCAGGCGAGGCTTTCTCG GCACCCGAGGCTCCAGCCTGATCTCGGTCTTATCCTGCGACCGGGCTGGTTCTGGCGG GTCGCCAGGGTGGGCGGCGGCCCCAGCCGGGCGCCCCGGCGGCAAGAGCGGCAGG CTGCGCCCCTGGCCCGCGCCTAGCCTGGGGAGAGAGCTGGGCGGGCGGCGGGAGCT GCTCTCGCGGGCCGCGGCCCTCGCCCTGGCTGCAACGGTAGGCGTTTCCCGGGCCGG ACGCGCGTGGGGGGCGGGGGCGGGGGCGGGGGCGAGGCCGCGGCGAGCAAAGTCC AGGCCCCTCTGCTGCAGCGCCCGCGCGTCCAGAGGCCCTGCCAGACACGCGCGAGGT TCGAGNNGAGAGAGGTCCGGGCGCGTCTGGCCTCGAAGGGAGACCCGGGACGTGGG GCGCGGGGCGGGAGTGGCCGGACCTCCACCCAGTGCCCCCGGGCCCCGCGACTCGT GCGCCGGGCCGCCGGAGAGGGTGTACTTGGTTCTGAGGCTGTGGTTTCTCCTCNNGCT GAGATGGATCTTGA

[0370] SEQ ID NO: 69 - CCL. LP1.coABCB11 CCATTGCATACGTTGTATCCATATCATAATATGTACATTTATATTGGCTCATGTCCAACAT TACCGCCATGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATT AGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTG GCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTA ACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCA CTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACG GTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGG CAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATC AATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGT CAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTC CGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGA GCTCGTTTAGTGAACCGGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCT GGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGTAGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGT CAGTGTGGAAAATCTCTAGCAGTGGCGCCCGAACAGGGACTTGAAAGCGAAAGGGAAA CCAGAGGAGCTCTCTCGACGCAGGACTCGGCTTGCTGAAGCGCGCACGGCAAGAGGC GAGGGGCGGCGACTGGTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGAAGGAGA GAGATGGGTGCGAGAGCGTCAGTATTAAGCGGGGGAGAATTAGATCGCGATGGGAAAA AATTCGGTTAAGGCCAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTATGGGCAA GCAGGGAGCTAGAACGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGT AGACAAATACTGGGACAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATC ATTATATAATACAGTAGCAACCCTCTATTGTGTGCATCAAAGGATAGAGATAAAAGACAC CAAGGAAGCTTTAGACAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCACAG CAAGCGGCCGCTGATCTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAGT GAATTATATAAATATAAAGTAGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGCA AAGAGAAGAGTGGTGCAGAGAGAAAAAAGAGCAGTGGGAATAGGAGCTTTGTTCCTTG GGTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACA GGCCAGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTG AGGCGCAACAGCATCTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAG AATCCTGGCTGTGGAAAGATACCTAAAGGATCAACAGCTCCTGGGGATTTGGGGTTGCT CTGGAAAACTCATTTGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTC TGGAACAGATTTGGAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTAC ACAAGCTTAATACACTCCTTAATTGAAGAATCGCAAAACCAGCAAGAAAAGAATGAACAA GAATTATTGGAATTAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACATAACAAATTGG CTGTGGTATATAAAATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTT TTGCTGTACTTTCTATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGA CCCACCTCCCAACCCCGAGGGGACCCGACAGGCCCGAAGGAATAGAAGAAGAAGGTG GAGAGAGAGACAGAGACAGATCCATTCGATTAGTGAACGGATCTCGACGGTATCGGTTA ACTTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGACA TAATAGCAACAGACATACAAACTAAAGAATTACAAAAACAAATTACAAAAATTCAAAATTT TATCGATCACGAGACTAGCCTCGAGGATATCCCCTAAAATGGGCAAACATTGCAAGCA GCAAACAGCAAACACACAGCCCTCCCTGCCTGCTGACCTTGGAGCTGGGGCAGAGGT CAGAGACCTCTCTGGGCCCATGCCACCTCCAACATCCACTCGACCCCTTGGAATTTCG GTGGAGAGGAGCAGAGGTTGTCCTGGCGTGGTTTAGGTAGTGTGAGAGGGGAATGAC TCCTTTCGGTAAGTGCAGTGGAAGCTGTACACTGCCCAGGCAAAGCGTCCGGGCAGC GTAGGCGGGCGACTCAGATCCCAGCCAGTGGACTTAGCCCCTGTTTGCTCCTCCGATA ACTGGGGTGACCTTGGTTAATATTCACCAGCAGCCTCCCCCGTTGCCCCTCTGGATCCA CTGCTTAAATACGGACGAGGACAGGGCCCTGTCTCCTCAGCTTCAGGCACCACCACTG ACCTGGGACAGTGAATCACCGGTGCCACCA TGA GCGA TTCTGTGA TCCTGCGGA GCA T CAA GA A GTTCGGCGAGGA AA ACGA CGGCTTCGA GA GCGA CAA GAGCTA CAACAA CG ACAAGAAGTCCCGGCTGCAGGACGAGAAGAAAGGCGACGGTGTCAGAGTGGGCTTC TTCCA GCTGTTCCGGTTCA GCAGCAGCACCGA CA TCTGGCTGA TGTTCGTGGGCA GC CTGTGCGCCTTTCTGCA CGGAATTGCTCA GCCTGGCGTGCTGCTGA TCTTCGGCACCA TGA CCGA CGTGTTCA TCGA CT A CGA CGTGGAA CTGCAA GA GCTGCA GA TCCCTGGCA A GGCCTGCGTGAACAA CA CCA TCGTGTGGA CCA A CA GCA GCCTGAA CCA GAA CA TGA CCA A CGGCACCA GA TGCGGCCTGCTGAATA TCGA GTCCGA GA TGA TCAAGTTCGCCA GCTA CTACGCCGGAATCGCCGTGGCCGTTCTGA TCA CCGGCTA CA TCCA GA TCTGCTT TTGGGTTA TCGCCGCTGCCA GA CA GA TCCA GA AG A TGCGGAAGTTCTA CTTCCGGCG GA TCA TGCGGA TGGAAA TCGGA TGGTTCGA CTGCAA CA GCGTGGGCGA GCTGAACAC CA GA TTCA GCGA CGA CA TCA A CAA GA TCA ACGA CGCCA TTGCCGACCA GA TGGCCCT GTTCA TCCA GCGGA TGA CCTCTA CCA TCTGCGGCTTTCTGCTGGGCTTTTTCA GA GGC TGGAAGCTGA CCCTGGTCA TCA TCA GCGTGTCCCCA CTGA TCGGA A TCGGA GCCGCC A CAA TCGGCCTGA GCGTGTCCAAGTTCA CCGA TTA CGA GCTGAAGGCCTA CGCCA A A GCTGGCGTGGTGGCCGATGAAGTGATCAGCTCCATGAGAACCGTGGCCGCCTTTGGC GGCGA GA AG A GA GA GGTGGAA CGCTA CGAGAAAAA CCTGGTGTTCGCCCA GA GA TG GGGCA TCA GAAA GGGCA TCGTGA TGGGA TTCTTCA CCGGCTTCGTGTGGTGCCTGA T CTTCCTGTGTTA CGCCCTGGCCTTTTGGTACGGCAGCACCCTGGTTCTGGA TGA GGGC GA GT A TA CCCCTGGAA CTCTGGTGCAGA TCTTTCTGA GCGTGA TCGTGGGCGCCCTGAA CCTGGG A TGCCTCTCCA TGCCTGGAA GCCTTTGCCA CA GGCAGA GCCGCCGCTA CCA GCA TCTTCGA GA CAATCGA CA GAAA GCCCA TCA TCGA CTGCA TGA GCGA GGA CG GCTA CAA GCTGGA CA GAA TCA A GGGCGA GA TCGA GTTCCACAA CGTGA CCTTTCA CT A CCCCA GCAGA CCCGAAGTGAA GA TCCTGAA CGA CCTGA A CA TGGTCA TCAA GCCCG GCGA GA TGA CA GCCCTCGTTGGA CCTTCTGGCGCCGGAAAA TCTA CA GCCCTGCA GC TGA TCCAGCGGTTCTA CGA TCCTTGCGAA GGCA TGGTCA CCGTGGACGGCCA CGA CA TCA GA TCTCTGAACA TCCA GTGGCTGCGGGACCA GA TCGGCA TCGTGGAA CAA GAGC CTGTGCTGTTCA GCACCA CAATCGCCGA GA AC A TCA GA TACGGCCGCGA GGA CGCCA CCA TGG A AG A TA TTGTGCA GGCCGCTAAA GA GGCCAA CGCCTA CAA CTTCA TCA TGG A CCTGCCTCA GCAGTTCGA CA CCCTCGTCGGA GAAGGCGGA GGA CAAA TGTCTGGCG GCCA GAAA CA GA GA GTGGCCA TTGCTA GA GCCCTGA TCA GAAA CCCCA AG A TCCTGC TGCTGGA CA TGGCCA CAAGCGCCCTGGA TA A TGA GA GCGA GGCCA TGGTGCAA GAG GTGCTGA GCA AG A TCCAGCACGGCCA CA CCA TCA TCAGTGTGGCCCA CA GA CTGA GC A CCGTGCGGGCTGCCGA TA CCA TCA TCGGA TTTGA GCA CGGCACCGCCGTGGAAA GA GGCACACACGAGGAACTGCTGGAACGGAAGGGCGTGTACTTCACCCTCGTGACACTG CA GA GCCA GGGCAA TCA GGCCCTGAA CGA A GA GGA CA TCAA GGA TGCCA CCGA GGA CGA CA TGCTGGCCA GAACCTTTAGCA GA GGCA GCTA CCA GGA CA GCCTGCGGGCCTC TA TCA GA CA GA GAAGCAA GA GCCA GCTGAGCTA CCTGGTGCA CGAA CCTCCA CTGGC TGTGGTGGA TCACAA GA GCACCTACGA GGAA GA TCGGAAGGACAAGGA CA TCCCCG TGCAAGAAGAAGTGGAACCCGCTCCTGTGCGGCGGATCCTGAAATTTTCTGCCCCTG AGTGGCCCTACATGCTCGTGGGATCTGTGGGAGCCGCCGTGAATGGAACAGTGACCC CTCTGTACGCCTTCCTGTTCTCCCA GA TCCTGGGCA CCTTCA GCA TCCCCG A CAAA GA GGAACAGCGGAGCCAGATCAATGGCGTGTGCCTGCTGTTTGTGGCTATGGGCTGCGT GTCCCTGTTTA CCCA GTTCCTGCAGGGA TA CGCCTTCGCCAAGA GCGGAGA GCTGCT GA CCA A GCGGCTGA GAAAGTTCGGCTTCA GA GCCA TGCTCGGCCA GGA TA TCGCTTG GTTTGA CGA CCTGA GAAA CA GCCCTGGCGCTCTGA CAA CCA GA CTGGCCA CA GA TGC TTCTCA GGTGCAAGGCGCTGCCGGCTCTCA GA TCGGAATGA TCGTGAA CA GCTTCA C CAA TGTGA CCGTGGCCA TGA TCA TTGCCTTTAGCTTCA GCTGGAAA CTGA GCCTCGTG A TCCTGTGCTTCTTCCCA TTTCTGGCCCTGTCCGGCGCTA CCCA GA CCAGAATGCTGA CA GGCTTCGCCA GCAGA GA CAA GCA GGCCCTGGAAA TGGTCGGA CA GA TCA CCAA C GAGGCCCTGAGCAACATCAGAACAGTGGCCGGCATCGGCAAAGAGCGGCGGTTTATT GA GGCTCTGGAAA CCGA GCTGGAAAAA CCCTTCAA GA CCGCCA TCCA GAAGGCCAAT A TCTA CGGCTTCTGCTTCGCCTTCGCTCAGTGCA TCA TGTTTA TCGCCAA CA GCGCCA GCTA TAGA TACGGCGGCTA CCTGA TCAGCAA TGA GGGCCTGCACTTCA GCTACGTGTT CCGCGTGA TCTCTGCCGTGGTGCTGTCTGCCA CA GCTCTGGGCA GA GCA TTCA GCTA CA CA CCCA GCTA CGCCA A GGCCAA GA TTA GCGCCGCCA GA TTCTTTCA GCTGCTGGA TA GA CA GCCTCCTA TCTCCGTGTA CAACACCGCTGGCGA GAAGTGGGA CAA CTTCCA GGGCAA GA TCGA CTTCGTGGA TTGCAAGTTCA CA TACCCCTCTCGGCCCGA CA GCCA GGTGCTGAATGGA CTGTCCGTGTCTA TCA GCCCCGGA CA GA CA CTGGCCTTTGTGGG A A GCTCTGGCTGCGGCAAAA GCA CCA GCA TCCA GCTGCTCGA GA GA TTCTACGA CCC CGA CCA GGGCAAAGTGA TGA TCGA CGGA CACGA CA GCAA GAAA GTGAA CGTGCAGT TCCTGA GA TCCAA CA TCGGGA TCGTGTCTCAA GAA CCCGTGCTGTTTGCCTGCTCCA T CA TGG A TA AC A TTA A GT A CGGCGA CAA CA CCAAA GAAA TCCCCA TGG AAA GAGTGA T CGCTGCCGCCAAA CA GGCCCAGCTGCA CGA TTTCGTGA TGA GCCTGCCTGA GAAGTA CGA GA CAAA CGTGGGCTCTCA GGGCA GCCA GCTGTCTA GA GGCGAAAA GCA GCGGA TCGCTA TCGCCA GA GCCA TCGTGCGGGA TCCTAA GA TTCTGCTCCTGGA TGAAGCCA CCAGCGCACTGGACACCGAGTCCGAAAAGACAGTGCAAGTGGCCCTGGACAAAGCC A GA GA GGGCAGAACCTGTA TCGTGA TCGCCCA CCGGCTGA GCA CAATCCAGAACGC CGA TA TTA TCGCCGTGA TGGCCCAGGGCGTCGTGA TCGAAAA GGGAA CCCA CGA AG A A CTGA TGGCTCA GAA GGGCGCCTA CT AC A A GCTCGTGA CAACAGGCA GCCCCA TCA G CTGAGTCGACAATCAACCTCTGGATTACAAAATTTGTGAAAGATTGACTGGTATTCTTAA CTATGTTGCTCCTTTTACGCTATGTGGATACGCTGCTTTAATGCCTTTGTATCATGCTATT GCTTCCCGTATGGCTTTCATTTTCTCCTCCTTGTATAAATCCTGGTTGCTGTCTCTTTATG AGGAGTTGTGGCCCGTTGTCAGGCAACGTGGCGTGGTGTGCACTGTGTTTGCTGACGC AACCCCCACTGGTTGGGGCATTGCCACCACCTGTCAGCTCCTTTCCGGGACTTTCGCTTTCCCCCTCCCTATTGCCACGGCGGAACTCATCGCCGCCTGCCTTGCCCGCTGCTGGAC AGGGGCTCGGCTGTTGGGCACTGACAATTCCGTGGTGTTGTCGGGGAAGCTGACGTCC TTTCCATGGCTGCTCGCCTGTGTTGCCACCTGGATTCTGCGCGGGACGTCCTTCTGCTA CGTCCCTTCGGCCCTCAATCCAGCGGACCTTCCTTCCCGCGGCCTGCTGCCGGCTCTG CGGCCTCTTCCGCGTCTTCGCCTTCGCCCTCAGACGAGTCGGATCTCCCTTTGGGCCG CCTCCCCGCCTGGAATTCGAGCTCGGTACCTTTAAGACCAATGACTTACAAGGCAGCTG TAGATCTTAGCCACTTTTTAAAAGAAAAGGGGGGACTGGAAGGGCTAATTCACTCCCAA CGAAGACAAGATCTGCTTTTTGCTTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGC CTGGGAGCTCTCTGGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTT GAGTGCTTCAAGTAGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTC AGACCCTTTTAGTCAGTGTGGAAAATCTCTAGCAGTAGTAGTTCATGTCATCTTATTATTC AGTATTTATAACTTGCAAAGAAATGAATATCAGAGAGTGAGAGGAACTTGTTTATTGCAG CTTATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTC ACTGCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGCTCTA GCTATCCCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATT CTCCGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCC TCTGAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGGACGTACCCAAT TCGCCCTATAGTGAGTCGTATTACGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGA CTGGGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCA GCTGGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCT GAATGGCGAATGGGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGT TACGCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTC TTCCCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCT CCCTTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGG GTGATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTG GAGTCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATC TCGGTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATG AGCTGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTTAGGT GGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCA AATATGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATAGCACCTAGAT CAAGAGACAGGATGAGGATCGTTTCGCATGATTGAACAAGATGGATTGCACGCAGGTTC TCCGGCCGCTTGGGTGGAGAGGCTATTCGGCTATGACTGGGCACAACAGACAATCGGC TGCTCTGATGCCGCCGTGTTCCGGCTGTCAGCGCAGGGGCGCCCGGTTCTTTTTGTCA AGACCGACCTGTCCGGTGCCCTGAATGAACTGCAAGACGAGGCAGCGCGGCTATCGTG GCTGGCCACGACGGGCGTTCCTTGCGCAGCTGTGCTCGACGTTGTCACTGAAGCGGG AAGGGACTGGCTGCTATTGGGCGAAGTGCCGGGGCAGGATCTCCTGTCATCTCACCTT GCTCCTGCCGAGAAAGTATCCATCATGGCTGATGCAATGCGGCGGCTGCATACGCTTG ATCCGGCTACCTGCCCATTCGACCACCAAGCGAAACATCGCATCGAGCGAGCACGTAC TCGGATGGAAGCCGGTCTTGTCGATCAGGATGATCTGGACGAAGAGCATCAGGGGCTC GCGCCAGCCGAACTGTTCGCCAGGCTCAAGGCGAGCATGCCCGACGGCGAGGATCTC GTCGTGACCCATGGCGATGCCTGCTTGCCGAATATCATGGTGGAAAATGGCCGCTTTTC TGGATTCATCGACTGTGGCCGGCTGGGTGTGGCGGACCGCTATCAGGACATAGCGTTG GCTACCCGTGATATTGCTGAAGAGCTTGGCGGCGAATGGGCTGACCGCTTCCTCGTGC TTTACGGTATCGCCGCTCCCGATTCGCAGCGCATCGCCTTCTATCGCCTTCTTGACGAG TTCTTCTGAATTATTAACGCTTACAATTTCCTGATGCGGTATTTTCTCCTTACGCATCTGT GCGGTATTTCACACCGCATCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTA TTTGTTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGACCAAAATCCCTTAACGT GAGTTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGA TCCTTTTTTTCTGCGCGTAATCTGCTGCTTGCAAACAAAAAAACCACCGCTACCAGCGGT GGTTTGTTTGCCGGATCAAGAGCTACCAACTCTTTTTCCGAAGGTAACTGGCTTCAGCA GAGCGCAGATACCAAATACTGTTCTTCTAGTGTAGCCGTAGTTAGGCCACCACTTCAAG AACTCTGTAGCACCGCCTACATACCTCGCTCTGCTAATCCTGTTACCAGTGGCTGCTGC CAGTGGCGATAAGTCGTGTCTTACCGGGTTGGACTCAAGACGATAGTTACCGGATAAG GCGCAGCGGTCGGGCTGAACGGGGGGGTCGTGCACACAGCCCAGCTTGGAGCGAAC GACCTACACCGAACTGAGATACCTACAGCGTGAGCTATGAGAAAGCGCCACGCTTCCCGAAGGGAGAAAGGCGGACAGGTATCCGGTAAGCGGCAGGGTCGGAACAGGAGAGCG CACGAGGGAGCTTCCAGGGGGAAACGCCTGGTATCTTTATAGTCCTGTCGGGTTTCGC CACCTCTGACTTGAGCGTCGATTTTTGTGATGCTCGTCAGGGGGGCGGAGCCTATGGA AAAACGCCAGCAACGCGGCCTTTTTACGGTTCCTGGCCTTTTGCTGGCCTTTTGCTCAC ATGTTCTTTCCTGCGTTATCCCCTGATTCTGTGGATAACCGTATTACCGCCTTTGAGTGA GCTGATACCGCTCGCCGCAGCCGAACGACCGAGCGCAGCGAGTCAGTGAGCGAGGAA GCGGAAGAGCGCCCAATACGCAAACCGCCTCTCCCCGCGCGTTGGCCGATTCATTAAT GCAGCTGGCACGACAGGTTTCCCGACTGGAAAGCGGGCAGTGAGCGCAACGCAATTAA TGTGAGTTAGCTCACTCATTAGGCACCCCAGGCTTTACACTTTATGCTTCCGGCTCGTAT GTTGTGTGGAATTGTGAGCGGATAACAATTTCACACAGGAAACAGCTATGACCATGATT ACGCCAAGCGCGCAATTAACCCTCACTAAAGGGAACAAAAGCTGGAGCTGCAAGCTTG G

[0371] Annotations:

[0372] • 74..657: CMV promoter - allows expression of the vector genome

[0373] • 658..7041: Vector genome

[0374] • 672..852: 5' LTR (truncated)- truncated 5' long terminal repeat (LTR) from HIV-1

[0375] • 899..1024: HIV-1 Ψ_ packaging signal of human immunodeficiency virus type 1

[0376] • 1517..1750: RRE_ The Rev response element (RRE) of HIV-1 allows for Rev-dependent mRNA export from the nucleus to the cytoplasm

[0377] • 2246..2363: cPPT / CTS_ central polypurine tract and central termination sequence of HIV- 1

[0378] • 2384..2841: LP1 promoter (shown bold)

[0379] • 2854..6819: Transgene (shown bold and in italics)

[0380] • 6826..7414: WPRE_ woodchuck hepatitis virus posttranscriptional regulatory element • 7501..7734: 3' LTR (AU3)_ self-inactivating 3' long terminal repeat (LTR) from HIV-1 • 7806..7927: SV40 poly(A) signal_ SV40 polyadenylation signal

[0381] • 7928..74: Part of the plasmid needed for bacterial work

[0382] SEQ ID NO: 70 - miR-coABCB4-LPaH, Version 3 backbone - coABCB11 CCATTGCATACGTTGTATCCATATCATAATATGTACATTTATATTGGCTCATGTCCAACAT TACCGCCATGTTGACATTGATTATTGACTAGTTATTAATAGTAATCAATTACGGGGTCATT AGTTCATAGCCCATATATGGAGTTCCGCGTTACATAACTTACGGTAAATGGCCCGCCTG GCTGACCGCCCAACGACCCCCGCCCATTGACGTCAATAATGACGTATGTTCCCATAGTA ACGCCAATAGGGACTTTCCATTGACGTCAATGGGTGGAGTATTTACGGTAAACTGCCCA CTTGGCAGTACATCAAGTGTATCATATGCCAAGTACGCCCCCTATTGACGTCAATGACG GTAAATGGCCCGCCTGGCATTATGCCCAGTACATGACCTTATGGGACTTTCCTACTTGG CAGTACATCTACGTATTAGTCATCGCTATTACCATGGTGATGCGGTTTTGGCAGTACATC AATGGGCGTGGATAGCGGTTTGACTCACGGGGATTTCCAAGTCTCCACCCCATTGACGT CAATGGGAGTTTGTTTTGGCACCAAAATCAACGGGACTTTCCAAAATGTCGTAACAACTC CGCCCCATTGACGCAAATGGGCGGTAGGCGTGTACGGTGGGAGGTCTATATAAGCAGA GCTCGTTTAGTGAACCGGGGTCTCTCTGGTTAGACCAGATCTGAGCCTGGGAGCTCTCT GGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAGTGCTTCAAGT AGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGACCCTTTTAGT CAGTGTGGAAAATCTCTAGCAGTGGCGCCCGAACAGGGACTTGAAAGCGAAAGGGAAA CCAGAGGAGCTCTCTCGACGCAGGACTCGGCTTGCTGAAGCGCGCACGGCAAGAGGC GAGGGGCGGCGACTGGTGAGTACGCCAAAAATTTTGACTAGCGGAGGCTAGAAGGAGA GAGATGGGTGCGAGAGCGTCAGTATTAAGCGGGGGAGAATTAGATCGCGATGGGAAAA AATTCGGTTAAGGCCAGGGGGAAAGAAAAAATATAAATTAAAACATATAGTATGGGCAA GCAGGGAGCTAGAACGATTCGCAGTTAATCCTGGCCTGTTAGAAACATCAGAAGGCTGT AGACAAATACTGGGACAGCTACAACCATCCCTTCAGACAGGATCAGAAGAACTTAGATC ATTATATAATACAGTAGCAACCCTCTATTGTGTGCATCAAAGGATAGAGATAAAAGACACCAAGGAAGCTTTAGACAAGATAGAGGAAGAGCAAAACAAAAGTAAGACCACCGCACAG CAAGCGGCCGCTGATCTTCAGACCTGGAGGAGGAGATATGAGGGACAATTGGAGAAGT GAATTATATAAATATAAAGTAGTAAAAATTGAACCATTAGGAGTAGCACCCACCAAGGCA AAGAGAAGAGTGGTGCAGAGAGAAAAAAGAGCAGTGGGAATAGGAGCTTTGTTCCTTG GGTTCTTGGGAGCAGCAGGAAGCACTATGGGCGCAGCGTCAATGACGCTGACGGTACA GGCCAGACAATTATTGTCTGGTATAGTGCAGCAGCAGAACAATTTGCTGAGGGCTATTG AGGCGCAACAGCATCTGTTGCAACTCACAGTCTGGGGCATCAAGCAGCTCCAGGCAAG AATCCTGGCTGTGGAAAGATACCTAAAGGATCAACAGCTCCTGGGGATTTGGGGTTGCT CTGGAAAACTCATTTGCACCACTGCTGTGCCTTGGAATGCTAGTTGGAGTAATAAATCTC TGGAACAGATTTGGAATCACACGACCTGGATGGAGTGGGACAGAGAAATTAACAATTAC ACAAGCTTAATACACTCCTTAATTGAAGAATCGCAAAACCAGCAAGAAAAGAATGAACAA GAATTATTGGAATTAGATAAATGGGCAAGTTTGTGGAATTGGTTTAACATAACAAATTGG CTGTGGTATATAAAATTATTCATAATGATAGTAGGAGGCTTGGTAGGTTTAAGAATAGTTT TTGCTGTACTTTCTATAGTGAATAGAGTTAGGCAGGGATATTCACCATTATCGTTTCAGA CCCACCTCCCAACCCCGAGGGGACCCGACAGGCCCGAAGGAATAGAAGAAGAAGGTG GAGAGAGAGACAGAGACAGATCCATTCGATTAGTGAACGGATCTCGACGGTATCGGTTA ACTTTTAAAAGAAAAGGGGGGATTGGGGGGTACAGTGCAGGGGAAAGAATAGTAGACA TAATAGCAACAGACATACAAACTAAAGAATTACAAAAACAAATTACAAAAATTCAAAATTT TATCGATCACGAGACTAGCCTCGAGGATATCCCATAGAGCCCACCGCATCCCCAGCAT GCCTGCTATTGTCTTCCCAATCCTCCCCCTTGCTGTCCTGCCCCACCCCACCCCCCAGA ATAGAATGACACCTACTCAGACAATGCGATGCAATTTCCTCATTTTATTAGGAAAGGACA GTGGGAGTGGCACCTTCCAGGGTCAAGGAAGGCACGGGGGAGGGGCAAACAACAGAT GGCTGGCAACTAGAAGGCACAGACGCGTGGGCCCCTGCAGTCGTACCGTGAGTAATAA TGCGATCGTCGTACCGTGAGTAATAATGCGCGGTTCGTACCGTGAGTAATAATGCGATC GTCGTACCGTGAGTAATAATGCGCTGCAGGTGTAGTGTTTCCTACTTTATGGAGTGATG TAGTGTTTCCTACTTTATGGACGGTTGTAGTGTTTCCTACTTTATGGAATCGTGTAGTGTT TCCTACTTTATGGAGGCTAGCCTAGGCGGGGAGGCGGCCCAAAGGGAGATCCGACTC GTCTGAGGGCGAAGGCGAAGACGCGGAAGAGGCCGCAGAGCCGGCAGCAGGCCGCG GGAAGGAAGGTCCGCTGGATTGAGGGCCGAAGGGACGTAGCAGAAGGACGTCCCGCG CAGAATCCAGGTGGCAACACAGGCGAGCAGCCATGGAAAGGACGTCAGCTTCCCCGAC AACACCACGGAATTGTCAGTGCCCAACAGCCGAGCCCCTGTCCAGCAGCGGGCAAGG CAGGCGGCGATGAGTTCCGCCGTGGCAATAGGGAGGGGGAAAGCGAAAGTCCCGGAA AGGAGCTGACAGGTGGTGGCAATGCCCCAACCAGTGGGGGTTGCGTCAGCAAACACA GTGCACACCACGCCACGTTGCCTGACAACGGGCCACAACTCCTCATAAAGAGACAGCA ACCAGGATTTATACAAGGAGGAGAAAATGAAAGCCATACGGGAAGCAATAGCATGATAC AAAGGCATTAAAGCAGCGTATCCACATAGCGTAAAAGGAGCAACATAGTTAAGAATACC AGTCAATCTTTCACAAATTTTGTAATCCAGAGGTTGATTGTCGACTCAGCTGATGGGGCT GCCTGTTGTCA CGA GCTTGTA GT A GGCGCCCTTCTGA GCCA TCA GTTCTTCGTGGGTT CCCTTTTCGA TCACGA CGCCCTGGGCCA TCA CGGCGA TA AT A TCGGCGTTCTGGA TTG TGCTCA GCCGGTGGGCGA TCA CGA TA CA GGTTCTGCCCTCTCTGGCTTTGTCCA GGGC CA CTTGCA CTGTCTTTTCGGA CTCGGTGTCCAGTGCGCTGGTGGCTTCA TCCA GGA GC AGA A TCTTAGGA TCCCGCACGA TGGCTCTGGCGA TAGCGA TCCGCTGCTTTTCGCCTC TA GA CA GCTGGCTGCCCTGA GA GCCCACGTTTGTCTCGTA CTTCTCA GGCA GGCTCA T CA CGAAA TCGTGCA GCTGGGCCTGTTTGGCGGCAGCGA TCA CTCTTTCCA TGGGGATT TCTTTGGTGTTGTCGCCGTA CTTAA TGTTA TCCA TGA TGGA GCA GGCA A A CA GCACGG GTTCTTGA GA CA CGA TCCCGA TGTTGGA TCTCA GGA A CTGCA CGTTCACTTTCTTGCT GTCGTGTCCGTCGA TCA TCA CTTTGCCCTGGTCGGGGTCGTA GAA TCTCTCGA GCAGC TGGA TGCTGGTGCTTTTGCCGCAGCCA GA GCTTCCCA CA A A GGCCA GTGTCTGTCCG GGGCTGA TA GA CA CGGA CAGTCCA TTCA GCA CCTGGCTGTCGGGCCGA GA GGGGTA T GTGAACTTGCAA TCCA CGAAGTCGA TCTTGCCCTGGA A GTTGTCCCA CTTCTCGCCA G CGGTGTTGTACACGGAGATAGGAGGCTGTCTATCCAGCAGCTGAAAGAATCTGGCGG CGCTAATCTTGGCCTTGGCGTAGCTGGGTGTGTAGCTGAATGCTCTGCCCAGAGCTGT GGCAGACAGCACCACGGCAGAGATCACGCGGAACACGTAGCTGAAGTGCAGGCCCT CA TTGCTGA TCA GGTA GCCGCCGTA TCTA TAGCTGGCGCTGTTGGCGA TAAA CA TGA T GCA CTGA GCGAA GGCGAA GCAGAAGCCGTA GA TA TTGGCCTTCTGGA TGGCGGTCTTGAA GGGTTTTTCCA GCTCGGTTTCCAGA GCCTCAA TA A A CCGCCGCTCTTTGCCGA TG CCGGCCA CTGTTCTGA TGTTGCTCA GGGCCTCGTTGGTGA TCTGTCCGA CCA TTTCCA GGGCCTGCTTGTCTCTGCTGGCGAAGCCTGTCAGCATTCTGGTCTGGGTAGCGCCGG A CA GGGCCA GAAA TGGGAAGAA GCA CA GGA TCA CGA GGCTCA GTTTCCAGCTGAAG CTAAA GGCAA TGA TCA TGGCCA CGGTCA CA TTGGTGAA GCTGTTCA CGA TCA TTCCGA TCTGA GA GCCGGCAGCGCCTTGCA CCTGA GAAGCA TCTGTGGCCA GTCTGGTTGTCA GA GCGCCA GGGCTGTTTCTCA GGTCGTCAAA CCA A GCGA TA TCCTGGCCGA GCA TGG CTCTGAAGCCGAACTTTCTCAGCCGCTTGGTCAGCAGCTCTCCGCTCTTGGCGAAGGC GTATCCCTGCAGGAACTGGGTAAACAGGGACACGCAGCCCATAGCCACAAACAGCA GGCACACGCCA TTGA TCTGGCTCCGCTGTTCCTCTTTGTCGGGGA TGCTGAAGGTGCC CA GGA TCTGGGA GAACA GGA A GGCGTA CA GAGGGGTCA CTGTTCCA TTCA CGGCGG CTCCCA CA GA TCCCA CGA GCA TGTAGGGCCA CTCA GGGGCA GA AAA TTTCA GGA TCC GCCGCACAGGAGCGGGTTCCACTTCTTCTTGCACGGGGATGTCCTTGTCCTTCCGATC TTCCTCGTAGGTGCTCTTGTGATCCACCACAGCCAGTGGAGGTTCGTGCACCAGGTAG CTCA GCTGGCTCTTGCTTCTCTGTCTGA TA GA GGCCCGCA GGCTGTCCTGGTAGCTGC CTCTGCTAAA GGTTCTGGCCA GCA TGTCGTCCTCGGTGGCA TCCTTGA TGTCCTCTTC GTTCA GGGCCTGA TTGCCCTGGCTCTGCAGTGTCA CGA GGGTGAAGTA CA CGCCCTT CCGTTCCAGCAGTTCCTCGTGTGTGCCTCTTTCCACGGCGGTGCCGTGCTCAAATCCG A TGA TGGTA TCGGCAGCCCGCACGGTGCTCA GTCTGTGGGCCA CA CTGA TGA TGGTG TGGCCGTGCTGGA TCTTGCTCA GCA CCTCTTGCA CCA TGGCCTCGCTCTCA TTA TCC A GGGCGCTTGTGGCCA TGTCCA GCAGCAGGA TCTTGGGGTTTCTGA TCA GGGCTCTA G CAA TGGCCA CTCTCTGTTTCTGGCCGCCA GA CA TTTGTCCTCCGCCTTCTCCGA CGA G GGTGTCGAACTGCTGAGGCAGGTCCATGATGAAGTTGTAGGCGTTGGCCTCTTTAGC GGCCTGCA CAA TA TCTTCCA TGGTGGCGTCCTCGCGGCCGTA TCTGA TGTTCTCGGCG A TTGTGGTGCTGAACAGCACAGGCTCTTGTTCCACGA TGCCGA TCTGGTCCCGCAGCC A CTGGA TGTTCA GA GA TCTGA TGTCGTGGCCGTCCACGGTGA CCA TGCCTTCGCAA G GA TCGTA GAACCGCTGGA TCA GCTGCAGGGCTGTA GA TTTTCCGGCGCCA GAA GGTC CAA CGA GGGCTGTCA TCTCGCCGGGCTTGA TGA CCA TGTTCA GGTCGTTCA GGA TCTT CACTTCGGGTCTGCTGGGGTAGTGAAAGGTCACGTTGTGGAACTCGATCTCGCCCTTG A TTCTGTCCA GCTTGTA GCCGTCCTCGCTCA TGCAGTCGA TGA TGGGCTTTCTGTCGA T TGTCTCGAAGATGCTGGTAGCGGCGGCTCTGCCTGTGGCAAAGGCTTCCAGGCATGG A GA GGCA TTTCCCA GGTTCA GGGCGCCCA CGA TCA CGCTCA GAAA GA TCTGCA CCA G AGTTCCAGGGGTATACTCGCCCTCATCCAGAACCAGGGTGCTGCCGTACCAAAAGGC CA GGGCGTAA CA CA GGA AG A TCAGGCA CCA CA CGAA GCCGGTGAAGAA TCCCA TCA CGA TGCCCTTTCTGA TGCCCCA TCTCTGGGCGAA CA CCA GGTTTTTCTCGTA GCGTTC CA CCTCTCTCTTCTCGCCGCCAAA GGCGGCCA CGGTTCTCA TGG A GCTGA TCACTTCA TCGGCCA CCACGCCA GCTTTGGCGTAGGCCTTCA GCTCGTAA TCGGTGAA CTTGGA C A CGCTCA GGCCGA TTGTGGCGGCTCCGA TTCCGA TCAGTGGGGA CA CGCTGA TGA TG A CCA GGGTCA GCTTCCA GCCTCTGAAAAA GCCCA GCAGAAA GCCGCA GA TGGTA GA GGTCA TCCGCTGGA TGAACAGGGCCA TCTGGTCGGCAA TGGCGTCGTTGA TCTTGTTG A TGTCGTCGCTGAA TCTGGTGTTCA GCTCGCCCA CGCTGTTGCAGTCGAA CCA TCCGA TTTCCATCCGCATGATCCGCCGGAAGTAGAACTTCCGCATCTTCTGGATCTGTCTGGC A GCGGCGA TA A CCCAAAA GCA GA TCTGGA TGTAGCCGGTGA TCA GAA CGGCCA CGG CGA TTCCGGCGTAGTA GCTGGCGAA CTTGA TCA TCTCGGA CTCGA TA TTCA GCAGGCC GCA TCTGGTGCCGTTGGTCA TGTTCTGGTTCA GGCTGCTGTTGGTCCA CA CGA TGGTG TTGTTCACGCAGGCCTTGCCAGGGATCTGCAGCTCTTGCAGTTCCACGTCGTAGTCGA TGA AC A CGTCGGTCA TGGTGCCGAAGA TCA GCA GCA CGCCA GGCTGA GCAA TTCCGT GCAGAAAGGCGCACAGGCTGCCCACGAACATCAGCCAGATGTCGGTGCTGCTGCTG AACCGGAACAGCTGGAAGAAGCCCACTCTGACACCGTCGCCTTTCTTCTCGTCCTGCA GCCGGGACTTCTTGTCGTTGTTGTAGCTCTTGTCGCTCTCGAAGCCGTCGTTTTCCTCG CCGAA CTTCTTGA TGCTCCGCA GGA TCA CA GAA TCGCTCA TGGTGGCACCGGTCTCAG CGGGAGGAGAAACCACAGCCTCAGAACCAAGTACACCCTCTCCGGCGGCCCGGCGC ACGAGTCGCGGGGCCCGGGGGCACTGGGTGGAGGTCCGGCCACTCCCGCCCCGCGC CCCACGTCCCGGGTCTCCCTTCGAGGCCAGACGCGCCCGGACCTCTCTCGGCTCGAA CCTCGCGCGTGTCTGGCAGGGCCTCTGGACGCGCGGGCGCTGCAGCAGAGGGGCCTGGACTTTGCTCGCCGCGGCCTCGCCCCCGCCCCCGCCCCCGCCCCCCACGCGCGTC CGGCCCGGGAAACGCCTACCGTTGCAGCCAGGGCGAGGGCCGCGGCCCGCGAGAG CAGCTCCCGCCGCCCGCCCAGCTCTCTCCCCAGGCTAGGCGCGGGCCAGGGGCGCA GCCTGCCGCTCTTGCCGCCGGGGCGCCCGGCTGGGGCCGCCGCCCACCCTGGCGAC CCGCCAGAACCAGCCCGGTCGCAGGATAAGACCGAGATCAGGCTGGAGCCTCGGGT GCCGAGAAAGCCTCGCCTGATGGGCCACCCCCTCGAGGCCTCTCACACTACCTAAAC CACGCCAGGACAACCTCTGCTCCTCTCCACCGAAATTCCAAGGGGTCGAGTGGATGT TGGAGGTGGCATGGGCCCAGAGAGGTCTCTGACCTCTGCCCCAGCTCCAAGGTCAGC AGGCAGGGAGGGCTGTGTGTTTGCTGTTTGCTGCTTGCAATGTTTGCCCATTTTAGGG GATATGCTTGAATTCGAGCTCGGTACCTTTAAGACCAATGACTTACAAGGCAGCTGTAG ATCTTAGCCACTTTTTAAAAGAAAAGGGGGGACTGGAAGGGCTAATTCACTCCCAACGA AGACAAGATCTGCTTTTTGCTTGTACTGGGTCTCTCTGGTTAGACCAGATCTGAGCCTG GGAGCTCTCTGGCTAACTAGGGAACCCACTGCTTAAGCCTCAATAAAGCTTGCCTTGAG TGCTTCAAGTAGTGTGTGCCCGTCTGTTGTGTGACTCTGGTAACTAGAGATCCCTCAGA CCCTTTTAGTCAGTGTGGAAAATCTCTAGCAGTAGTAGTTCATGTCATCTTATTATTCAGT ATTTATAACTTGCAAAGAAATGAATATCAGAGAGTGAGAGGAACTTGTTTATTGCAGCTT ATAATGGTTACAAATAAAGCAATAGCATCACAAATTTCACAAATAAAGCATTTTTTTCACT GCATTCTAGTTGTGGTTTGTCCAAACTCATCAATGTATCTTATCATGTCTGGCTCTAGCT ATCCCGCCCCTAACTCCGCCCATCCCGCCCCTAACTCCGCCCAGTTCCGCCCATTCTC CGCCCCATGGCTGACTAATTTTTTTTATTTATGCAGAGGCCGAGGCCGCCTCGGCCTCT GAGCTATTCCAGAAGTAGTGAGGAGGCTTTTTTGGAGGCCTAGGGACGTACCCAATTCG CCCTATAGTGAGTCGTATTACGCGCGCTCACTGGCCGTCGTTTTACAACGTCGTGACTG GGAAAACCCTGGCGTTACCCAACTTAATCGCCTTGCAGCACATCCCCCTTTCGCCAGCT GGCGTAATAGCGAAGAGGCCCGCACCGATCGCCCTTCCCAACAGTTGCGCAGCCTGAA TGGCGAATGGGACGCGCCCTGTAGCGGCGCATTAAGCGCGGCGGGTGTGGTGGTTAC GCGCAGCGTGACCGCTACACTTGCCAGCGCCCTAGCGCCCGCTCCTTTCGCTTTCTTC CCTTCCTTTCTCGCCACGTTCGCCGGCTTTCCCCGTCAAGCTCTAAATCGGGGGCTCCC TTTAGGGTTCCGATTTAGTGCTTTACGGCACCTCGACCCCAAAAAACTTGATTAGGGTG ATGGTTCACGTAGTGGGCCATCGCCCTGATAGACGGTTTTTCGCCCTTTGACGTTGGAG TCCACGTTCTTTAATAGTGGACTCTTGTTCCAAACTGGAACAACACTCAACCCTATCTCG GTCTATTCTTTTGATTTATAAGGGATTTTGCCGATTTCGGCCTATTGGTTAAAAAATGAGC TGATTTAACAAAAATTTAACGCGAATTTTAACAAAATATTAACGCTTACAATTTAGGTGGC ACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTGTTTATTTTTCTAAATACATTCAAATA TGTATCCGCTCATGAGACAATAACCCTGATAAATGCTTCAATAATAGCACCTAGATCAAG AGACAGGATGAGGATCGTTTCGCATGATTGAACAAGATGGATTGCACGCAGGTTCTCCG GCCGCTTGGGTGGAGAGGCTATTCGGCTATGACTGGGCACAACAGACAATCGGCTGCT CTGATGCCGCCGTGTTCCGGCTGTCAGCGCAGGGGCGCCCGGTTCTTTTTGTCAAGAC CGACCTGTCCGGTGCCCTGAATGAACTGCAAGACGAGGCAGCGCGGCTATCGTGGCT GGCCACGACGGGCGTTCCTTGCGCAGCTGTGCTCGACGTTGTCACTGAAGCGGGAAG GGACTGGCTGCTATTGGGCGAAGTGCCGGGGCAGGATCTCCTGTCATCTCACCTTGCT CCTGCCGAGAAAGTATCCATCATGGCTGATGCAATGCGGCGGCTGCATACGCTTGATC CGGCTACCTGCCCATTCGACCACCAAGCGAAACATCGCATCGAGCGAGCACGTACTCG GATGGAAGCCGGTCTTGTCGATCAGGATGATCTGGACGAAGAGCATCAGGGGCTCGCG CCAGCCGAACTGTTCGCCAGGCTCAAGGCGAGCATGCCCGACGGCGAGGATCTCGTC GTGACCCATGGCGATGCCTGCTTGCCGAATATCATGGTGGAAAATGGCCGCTTTTCTGG ATTCATCGACTGTGGCCGGCTGGGTGTGGCGGACCGCTATCAGGACATAGCGTTGGCT ACCCGTGATATTGCTGAAGAGCTTGGCGGCGAATGGGCTGACCGCTTCCTCGTGCTTT ACGGTATCGCCGCTCCCGATTCGCAGCGCATCGCCTTCTATCGCCTTCTTGACGAGTTC TTCTGAATTATTAACGCTTACAATTTCCTGATGCGGTATTTTCTCCTTACGCATCTGTGCG GTATTTCACACCGCATCAGGTGGCACTTTTCGGGGAAATGTGCGCGGAACCCCTATTTG TTTATTTTTCTAAATACATTCAAATATGTATCCGCTCATGACCAAAATCCCTTAACGTGAG TTTTCGTTCCACTGAGCGTCAGACCCCGTAGAAAAGATCAAAGGATCTTCTTGAGATCCT TTTTTTCTGCGCGTAATCTGCTGCTTGCAAAC...

Claims

Claims1. A promoter sequence comprising at least a portion of a ABCB4 promoter sequence or reverse complement thereof, wherein the splice sites of the ABCB4 promoter are mutated.

2. The promoter sequence of claim 1, wherein the promoter sequence comprises a portion of the human ABCB4 promoter sequence or reverse complement thereof.

3. The promoter sequence of any preceding claim, wherein at least a portion of the promoter sequence comprises at least 95 % sequence identity, or at least 99% sequence identity to SEQ ID NO: 1, optionally at least 95 % sequence identity, or at least 99% sequence identity to SEQ ID NO: 3.

4. The promoter sequence of any preceding claim, wherein at least a portion of the promoter sequence comprises at least 95 % sequence identity, or at least 99% sequence identity to SEQ ID NO: 2, optionally at least 95 % sequence identity, or at least 99% sequence identity to SEQ ID NO: 4.

5. A lentiviral vector sequence comprising the promoter sequence of any one of claims 1- 4 which is operably linked with a transgene.

6. A lentiviral vector sequence comprising the promoter sequence of any one of claims 1- 4, wherein the promoter sequence and the transgene are in reverse orientation to the direction of transcription of the lentiviral vector sequence.

7. The promoter sequence of any one of claims 1-4, or the lentiviral vector sequence of any one of claims 5-6, further comprising an enhancer sequence, optionally an HCR enhancer, preferably as defined in SEQ ID NO: 5 or the reverse complement thereof, and further preferably wherein the lentiviral vector sequence comprises a sequence with at least 95% sequence identity, or at least 99% sequence identity or is identical to SEQ ID NO: 6 or SEQ ID NO: 7.

8. The lentiviral vector sequence of any one of claims 5-7, wherein the transgene encodes for a liver protein, optionally a liver transmembrane protein.

9. The lentiviral vector sequence of any one of claims 5-8, wherein the transgene encodes for an ATB binding cassette transporter protein, preferably ABC B4 orABCBH.

10. The lentiviral vector sequence of any one of claims 5-9, wherein the transgene encodes ABCB4.

11. The lentiviral vector sequence of claim 10, wherein the transgene(i) encodes human ABCB4 and / or(ii) comprises a sequence with at least 95% sequence identity, or 100% sequence identity to SEQ ID NO: 8 and / or(iii) coding sequence has at least 95% sequence identity, or 100% sequence identity to SEQ ID NO: 43.

12. The lentiviral vector sequence of claim 10, wherein the transgene(i) is a codon-optimised ABCB4 gene, and / or(ii) comprises a sequence with at least 95% sequence identity, or 100% sequence identity to SEQ ID NO: 9, and / or(iii) coding sequence has at least 95% sequence identity, or 100% sequence identity to SEQ ID NO: 44.

13. The lentiviral vector sequence of claim 9, wherein the transgene encodes ABCB11, optionally wherein the transgene comprises a sequence with at least 95% sequence identity, or 100% sequence identity to SEQ ID NO: 72 or SEQ ID NO: 73.

14. The lentiviral vector sequence of any one of claims 5-13, further comprising one or more miRNA sequence, preferably one or more repeat miRNA binding sequences.

15. The lentiviral vector sequence of claim 14, wherein the one or more miRNA sequences comprises one or more of miR-142 and / or one or more of miR-126-3pT, optionally comprising a sequence with at least 95% sequence identity, or 100% sequence identity to SEQ ID NO: 10-14.

16. The lentiviral vector sequence of any one of claims 5-15, wherein the lentiviral vector sequence is based on a human immunodeficiency virus (HIV), preferably HIV-117. The lentiviral vector sequence of any one of claims 5-16, comprising at least 95 % sequence identity, or at least 99% sequence identity to SEQ ID NO: 15 or SEQ ID NO: 16 or SEQ ID NO: 71.

18. A lentiviral vector comprising the lentiviral vector sequence as defined in any one of claims 5-17.

19. The lentiviral vector sequence of any one of claims 5-17, or the lentiviral vector of claim 18, for use in therapy.

20. The lentiviral vector sequence of any one of claims 8-17, or the lentiviral vector of claim 18, for use in the treatment of a liver disease.

21. The lentiviral vector sequence of any one of claims 8-12 or 14-17, or the lentiviral vector of claim 18, for use in the treatment of cholestasis.

22. The lentiviral vector sequence of any one of claims 8-12 or 14-17, or the lentiviral vector of claim 18, for use in the treatment of progressive familiar intrahepatic cholestasis (PFIC), preferably PFIC3, wherein the transgene encodes ABCB4.

23. The lentiviral vector sequence of any one of claims 10-12 or 14-17, or the lentiviral vector of claim 18, for use in the treatment of a disease or disorder associated with mutated ABCB4 and / or aberrant expression of ABCB4, optionally wherein the disease or disorder is PFIC3, intrahepatic cholestasis of pregnancy type 3 (ICP3), gallbladder disease 1 (GBD1), or low phospholipid associated cholelithiasis.

24. A host cell comprising the lentiviral vector sequence of any one of claims 5-17 or the lentiviral vector of claim 18.

25. A composition comprising a lentiviral vector sequence according to any one of claims 5-17, or the lentiviral vector of claim 18, and a pharmaceutically acceptable carrier.

26. Use of the promoter sequence as defined in any one of claims 1-4, or the lentivirus vector sequence as defined in any one of claims 5-17, to improve lentiviral titre.

27. A plasmid comprising the lentiviral vector sequence as defined in any of one of claims 5-17.

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